Fermentations

J Agric Food Chem, 2003 Jan 15, 51(2), 424 - 32
Chemical and sensorial aroma characterization of freshly distilled Calvados . 1 . Evaluation of quality and defects on the basis of key odorants by olfactometry and sensory analysis; Guichard H et al.; Eight freshly distilled samples of Calvados, a fermented and distilled apple juice, were analyzed by sensory evaluation and direct injection GC to determine the composition of higher alcohols, esters, and aldehydes . The composition determined by direct injection was tentatively related to sensory descriptors . Esters have a probable maximum level around 500 g/hl of pure alcohol (PA) . This level also corresponds to the threshold of the main ester constituent, ethyl acetate . A high ratio of esters to ethyl acetate seems to be of prime importance for good quality . Total aldehydes, with a maximum level between 8 and 11 g/hl of PA and mainly comprising acetal (maximum between 5 and 9 g/hl of PA), were related to a "green" descriptor . Higher alcohols do not have a direct impact on quality, but other volatile compounds with a positive impact on flavor should probably be present at a high level . As overall quality was not well related to sensory quality, it was necessary to perform more a precise analysis to determine the key odorants . The Calvados samples were thus extracted using pentane . Gas chromatography, employing both a flame ionization detector and an olfactometry port, was used to analyze the obtained extracts . Seventy-one odors were detected and distributed according to Calvados quality determined by sensory evaluation . Nineteen odors common to all Calvados samples constituted the "skeleton" of the aroma . Twenty-eight odors were specific to a quality class: 6 for good quality, 4 for neutral, and 18 for defective . Twenty-four other odors had either too low an odor impact or no evident specificity.

Biosci Rep, 2002 Jun-Aug, 22(3-4), 421 - 30
F0 cysteine, bCys21, in the Escherichia coli ATP synthase is involved in regulation of potassium uptake and molecular hydrogen production in anaerobic conditions; Mnatsakanyan N et al.; The single cysteine in the b subunit of the membranous F0 sector and the 19 cysteines in extramembranous F1 sector of the Escherichia coli ATP synthase were replaced by alanine . When cells were grown under anaerobic conditions on glucose, the kcat for ATP hydrolysis of membrane vesicles containing the bCys21Ala mutant enzyme, but not enzymes with other cysteine replacements, was lower, while ATP-driven H+ pumping was unchanged . However, the ATP-dependent increase in the number of accessible thiol groups in membrane vesicles was negated . Furthermore, K+ uptake and molecular hydrogen production by whole cells and protoplasts was greatly decreased . These results indicate a role for the F0 subunit bCys21 in the functionality of F0F1 and coupling to other membranous activities under fermentative conditions.

J Am Diet Assoc, 1998 Feb, 98(2), 182 - 6, 189; quiz 187-8
Nutrition and oral health guidelines for pregnant women, infants, and children; Fitzsimons D et al.; Good oral health care and nutrition during pregnancy, infancy, and childhood are essential but often overlooked factors in the growth and development of the teeth and oral cavity . Pregnant women and parents and caregivers of infants and children often receive little guidance about proper preventive dental and oral health care, including fluoride and dietary measures . Pregnant women can maintain their health through proper diet, good oral hygiene, and appropriate use of fluoride . An adequate diet during gestation is important for optimal oral development of the fetus . To promote good oral health in infancy, caregivers need to provide the infant with appropriate amounts of fluoride in addition to a healthful diet . As the teeth erupt into the mouth, the caregiver needs to clean the teeth thoroughly on a daily basis . When solid foods are introduced in later infancy, it is also important to limit the frequency of caries-promoting fermentable carbohydrates between meals . Good oral hygiene habits and dietary practices that emphasize minimum exposure to retentive, fermentable carbohydrates; use of fluoridated water; and a varied, balanced diet should continue throughout childhood to set the stage for optimal oral health for a lifetime.

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi, 1998 Jun, 12(2), 139 - 42
{The study on purification and characterization of HIV-1 reverse transcriptase from a recombinant strain of E . coli}; Jia H et al.; Of the HIV proteins, reverse transcriptase(RT) has been probably the most useful target protein for screening and designing of its specific inhibitors . Because retroviral replication is absolutely dependent on both the RNase H and the polymerase function of RT and, so far as is now known, RT does not play a direct role in the life cycle of a normal cell . Under suitable fermentation conditions in our experiments, HIV-1 RT was highly expressed in E . coli JM109(pKRT-2)* by inducing the trc promoter with isopropyl-beta-Dthiogalactopyranoside(IPTG) . 1 . 1 mg of purified RT was obtained from one liter culture of bacteria by DEAE-cellulose and phosphaellulose chromatography . SDS-PAGE analysis of the purified RT showed two major protein bands of 66 kD and 51 kD, indicating that the purified RT was a heterodimer composed of two subunits . Results of enzyme assay showed that the purified RT had high activity(1.4 x 10(4) umit/mg) . We also improved the reaction system of enzyme assay . The effect of PFA on HIV-1 RT was determined with the improved enzyme assay and the mechanism of inhibition was non-competitive with respect to substrate consistent with the reports of Dr . Bo Oberg . This suggests that the purified HIV-1 RT by this simple method can be applied to the anti HIV-1-drug screening . (*E . coli JM109(pKRT2) was obtained from NIAID, NIH; pKRT2 from Dr . Richard D'Aquila and Dr . William C . Summers.)

Biotechnol Bioeng, 2003 Mar 5, 81(5), 525 - 34
Effect of deletion of chitin synthase genes on mycelial morphology and culture viscosity in Aspergillus oryzae; Muller C et al.; The objective of this study was to quantify the effect of disrupting two chitin synthases, chsB and csmA, on the morphology and rheology during batch cultivation of Aspergillus oryzae . The rheological properties were characterized in batch cultivations at different biomass concentrations (from 3.4-22.5 g kg(-1) biomass) and the power-law model adequately described the rheological properties . In the cultivations there were pellets, clumps, and freely dispersed hyphal elements . The different morphological fractions were quantified using image analysis . The apparent viscosity of the fermentation broth was significantly affected by the biomass concentration, the morphology, and also by pH . The chsB disruption strain had lower consistency index K values for all biomass concentrations investigated, which is a desirable trait for industrial Aspergillus fermentations .

J Nutr, 2003 Jan, 133(1), 1 - 4
Effects of dietary fibers on magnesium absorption in animals and humans; Coudray C et al.; There is overwhelming evidence that dietary fibers are an important component of human and animal diets and play an important role in human health . Because dietary fibers and some associated substances, such as phytate, have in vitro mineral-binding capacities, they have been thought to impair absorption of minerals such as calcium, iron and zinc, although magnesium absorption seems to be less affected . Indeed, the effect of dietary fibers depends largely on their own nature and characteristics, and also on mineral homeostasis . In 1977 it was observed that resistant starch, a fermentable dietary fiber, could improve Mg absorption in rats . More recently, attention has been focused on other fermentable substrates such as inulin and oligo- or polysaccharides, for their potential prebiotic and health effects . Studies conducted on different types of fermentable carbohydrates have confirmed their beneficial effect on Mg absorption in different animal species . The majority of these studies have also sought to determine the effects of fibers on other minerals such as calcium, iron and zinc . In contrast to the studies with Mg, these studies did not show a consistent effect on the absorption of these minerals . This is due to the particularities of sites and mechanism of Mg absorption . To date, four human studies have been carried out that generally confirmed the enhancing effect of fermentable oligo- or polysaccharides on Mg absorption.

Appl Environ Microbiol, 2003 Jan, 69(1), 227 - 32
Purification and characterization of a prolyl aminopeptidase from Debaryomyces hansenii; Bolumar T et al.; A prolyl aminopeptidase (PAP) (EC 3.4.11.5) was isolated from the cell extract of Debaryomyces hansenii CECT12487 . The enzyme was purified by selective fractionation with protamine and ammonium sulfate, followed by two chromatography steps, which included gel filtration and anion-exchange chromatography . The PAP was purified 248-fold, with a recovery yield of 1.4% . The enzyme was active in a broad pH range (from 5 to 9.5), with pH and temperature optima at 7.5 and 45 degrees C . The molecular mass was estimated to be around 370 kDa . The presence of inhibitors of serine and aspartic proteases, bestatin, puromycin, reducing agents, chelating agents, and different cations did not have any effect on the enzyme activity . Only iodoacetate, p-chloromercuribenzoic acid, and Hg(2+), which are inhibitors of cysteine proteases, markedly reduced the enzyme activity . The K(m) for proline-7-amido-4-methylcoumarin was 40 micro M . The enzyme exclusively hydrolyzed N-terminal-proline-containing substrates . This is the first report on the identification and purification of this type of aminopeptidase in yeast, which may contribute to the scarce knowledge about D . hansenii proteases and their possible roles in meat fermentation.

Appl Environ Microbiol, 2003 Jan, 69(1), 113 - 21
Oxygen consumption by anaerobic Saccharomyces cerevisiae under enological conditions: effect on fermentation kinetics; Rosenfeld E et al.; The anaerobic growth of the yeast Saccharomyces cerevisiae normally requires the addition of molecular oxygen, which is used to synthesize sterols and unsaturated fatty acids (UFAs) . A single oxygen pulse can stimulate enological fermentation, but the biochemical pathways involved in this phenomenon remain to be elucidated . We showed that the addition of oxygen (0.3 to 1.5 mg/g {dry mass} of yeast) to a lipid-depleted medium mainly resulted in the synthesis of the sterols and UFAs required for cell growth . However, the addition of oxygen during the stationary phase in a medium containing excess ergosterol and oleic acid increased the specific fermentation rate, increased cell viability, and shortened the fermentation period . Neither the respiratory chain nor de novo protein synthesis was required for these medium- and long-term effects . As de novo lipid synthesis may be involved in ethanol tolerance, we studied the effect of oxygen addition on sterol and UFA auxotrophs (erg1 and ole1 mutants, respectively) . Both mutants exhibited normal anaerobic fermentation kinetics . However, only the ole1 mutant strain responded to the oxygen pulse during the stationary phase, suggesting that de novo sterol synthesis is required for the oxygen-induced increase of the specific fermentation rate . In conclusion, the sterol pathway appears to contribute significantly to the oxygen consumption capacities of cells under anaerobic conditions . Nevertheless, we demonstrated the existence of alternative oxygen consumption pathways that are neither linked to the respiratory chain nor linked to heme, sterol, or UFA synthesis . These pathways dissipate the oxygen added during the stationary phase, without affecting the fermentation kinetics.

J Dairy Sci, 2002 Dec, 85(12), 3454 - 61
Effects of a monensin controlled-release capsule or premix on attenuation of subacute ruminal acidosis in dairy cows; Mutsvangwa T et al.; The effects of monensin, administered either as a controlled release capsule (CRC) or a premix, on attenuating grain-induced subacute ruminal acidosis (SARA) and on ruminal fermentation characteristics in Holstein cows receiving a total mixed ration were investigated in two experiments . In both experiments, six multiparous, rumen-fistulated Holstein cows were used in a two-treatment, two-period crossover design with 6-wk periods . In Experiment 1, treatments were either a monensin CRC or a placebo CRC . In Experiment 2, treatments were either a monensin premix or a placebo premix . In both experiments, at the beginning of wk 4 SARA was induced in experimental cows for a 10-d period with a grain challenge model, and ruminal pH was measured continuously using indwelling pH probes . The administration of monensin either as a CRC or a premix had no effect on ruminal pH characteristics . Neither monensin CRC nor premix had an effect on ruminal volatile fatty acid concentrations, but reduced the acetate:propionate ratio . Monensin premix-treated cows were observed to have increased milk yield, largely as a result of a higher dry matter intake in monensin-treated cows compared to control cows . Milk fat content and yield were lower in monensin-treated cows compared to placebo-treated cows during SARA . In conclusion, there is no evidence that monensin was efficacious in raising ruminal pH during SARA under the conditions employed in this study.

J Dairy Sci, 2002 Dec, 85(12), 3411 - 9
Effects of cultivars on ensiling characteristics, chemical composition, and ruminal degradability of pea silage; Mustafa AF et al.; A study was conducted to determine the effects of cultivar on ensiling characteristics, chemical composition and ruminal nutrient degradability of pea (Pisum sativum L.) silage . The cultivars evaluated were Lenca (L), Carneval (C), and Delta (D) . Peas were field-grown and forage was harvested and ensiled in mini-silos for 0, 2, 4, 8, 16, and 70 d . The ensiled forage of all cultivars went through a rapid fermentation with a sharp reduction in pH during the first 2 days of ensiling . Extensive proteolysis took place between 0 and 2 d as indicated by a reduction in true protein and neutral detergent insoluble protein (NDICP) and an increase in nonprotein nitrogen . Chemical analysis of the 70 d silage showed that cultivar L contained higher neutral detergent fiber (NDF) and acid detergent fiber and lower starch levels than C and D . Crude protein was highest for C (20.5% DM), intermediate for D (19.0% DM) and lowest for L (17.9% DM) . Distribution of protein fractions showed that L contains lower soluble protein and higher NDICP levels than the other two pea cultivars . However, no difference in acid detergent insoluble protein levels was observed between the three cultivars . Results of the in situ incubation experiment indicated that L had lower ruminal DM (69.2 vs 74.0%) and CP (84.1 vs 90.6%) degradabilities than C or D . However, ruminal degradability of NDF was similar among the three cultivars (average of 32.9%) . It was concluded that chemical composition and ruminal nutrient degradability of pea silage are significantly influenced by cultivars.

J Dairy Sci, 2002 Dec, 85(12), 3336 - 51
Effect of protein source on amino acid supply, milk production, and metabolism of plasma nutrients in dairy cows fed grass silage; Korhonen M et al.; This study conducted according to a 4 x 4 Latin square with 28 d periods and four ruminally cannulated Finnish Ayrshire cows investigated the effect of protein supplements differing in amino acid (AA) profile and rumen undegradable protein content on postruminal AA supply and milk production . Mammary metabolism of plasma AA and other nutrients were also studied . The basal diet (Control; 13.4% crude protein) consisted of grass silage and barley in a ratio of 55:45 on a dry matter basis . The other three isonitrogenous diets (17.0% crude protein) were control + fishmeal (FM), control + soybean meal (SBM), and control + corn gluten meal (CGM) . The protein supplements replaced portions of dry matter of the control diet maintaining the silage to barley ratio constant for all diets . Dry matter intake was limited to 95% of the preexperimental ad libitum intake and was similar (mean 19.8 kg/d dry matter) across the diets . Protein supplements increased milk, lactose, and protein yields but did not affect yields of energy-corrected milk or milk fat . Milk protein yield response was numerically lowest for diet SBM . Protein supplements increased milk protein concentration but decreased milk fat and lactose concentrations . Microbial protein synthesis and rumen fermentation parameters were similar across the diets, except for an increased rumen ammonia concentration for diets supplemented with protein feeds . Protein supplements increased N intake, ruminal organic matter and N, and total tract organic matter, N, and neutral detergent fiber digestibilities . Protein supplements also increased N and AA flows into the omasum, with SBM giving the lowest and CGM the highest flows . This was associated with an unchanged microbial N flow and a higher undegraded dietary N flow . The omasal flows of individual AA reflected differences in total N flow and AA profile of the experimental diets . Differences in AA flows did not always reflect plasma AA concentrations . The results indicated that AA supply of dairy cows fed a grass silage-cereal diet can be manipulated using protein supplements differing in ruminal protein degradability and AA profile . Lower milk production response to SBM than that to FM and CGM appeared to be related mainly to lower N and AA supplies arising from a high ruminal protein degradability of SBM . Histidine appeared to be the first limiting AA for milk protein synthesis on the control diet . Mammary gland may regulate AA uptake according to requirements.

Ann Bot (Lond), 2003 Jan, 91 Spec No, 279 - 90
Dynamic aspects of alcoholic fermentation of rice seedlings in response to anaerobiosis and to complete submergence: relationship to submergence tolerance; Boamfa EI et al.; Rice plants are severely damaged by complete submergence . This is a problem in rice farming and could be the result, in part, of tissue anoxia imposed by a reduced availability of oxygen . To investigate this possibility we monitored alcoholic fermentation products as markers for tissue anaerobiosis using sensitive laser-based spectroscopy able to sense ethanol and acetaldehyde down to 3 nl l(-1) and 0.1 nl l(-1), respectively . Acetaldehyde emission began within 0.5 h of imposing an oxygen-free gas phase environment followed closely by ethanol . As treatment progressed, ethanol output increased and came to exceed acetaldehyde emission as this stabilized considerably after approx . 3 h . On re-entry of air, a sharp post-anaerobic peak of acetaldehyde production was observed . This was found to be diagnostic of a preceding anoxic episode of 0.5 h or more . When anaerobiosis was lengthened by up to 14 h, the size of the post-anaerobic acetaldehyde outburst increased . After de-submergence from oxygen-free water, a similarly strong but slower post-anaerobic acetaldehyde upsurge was seen, which was accompanied by an increase in ethanol emission . Light almost, but not completely, eliminated fermentation in anaerobic surroundings and also the post-anaerobic or post-submergence peaks in acetaldehyde production . All photosynthetically generated oxygen was consumed within the plant . There was no substantial difference in acetaldehyde and ethanol output between FR13A and the less submergence-tolerant line CT6241 under any submergence treatment . In some circumstances, submergence damaged CT6241 more than FR13A even in the absence of vigorous fermentation . We conclude that oxygen deprivation may not always determine the extent of damage caused to rice plants by submergence under natural conditions.

Ann Bot (Lond), 2003 Jan, 91 Spec No, 195 - 204
Assessment of enzyme induction and aerenchyma formation as mechanisms for flooding tolerance in Trifolium subterraneum 'Park'; Aschi-Smiti S et al.; The objective of this study was to evaluate the role of enzyme induction and aerenchyma formation in prolonged tolerance to soil flooding in a variety of underground clover (Trifolium subterraneum 'Park') previously selected for resistance . Seedlings were grown in hydroponic tanks, initially with aeration for 3 weeks and subsequently in the absence of aeration for up to 3 weeks . After 1 h in the absence of aeration, the oxygen concentration in the hydroponic medium had decreased to 1.5 % . During the 3 weeks of extreme oxygen deficiency, primary roots died and were replaced by considerable numbers of adventitious roots . Activities of many glycolytic and fermentative enzymes increased in adventitious roots . Excised adventitious roots were capable of immediate induction of ethanol in the absence of lactate production, in association with energy charge higher than that in excised roots of aerobically maintained controls . Energy charge was even higher when measured in adventitious roots in planta . Interestingly, haemoglobin protein could be correlated with energy charge . Aerenchyma was readily visualized in adventitious roots by optical microscopy of longitudinal and transverse sections . We conclude that avoidance of root anoxia via aerenchyma is the major mechanism for prolonged root tolerance in Trifolium subterraneum 'Park'.

Ann Bot (Lond), 2003 Jan, 91 Spec No, 155 - 72
Functional electron microscopy in studies of plant response and adaptation to anaerobic stress; Vartapetian BB et al.; This article reviews the contribution made by functional electron microscopy towards identifying and understanding the reactions of plant roots and shoots to anaerobic stress . Topics examined include: (1) unexpected hypersensitivity, rather than hyper-resistance, to anoxia of root tips of flooding-tolerant plants; (2) protective, rather than damaging, effects of a stimulated energy metabolism (glycolysis and fermentation) under anaerobic conditions; (3) the concept of two main strategies of plant adaptation to anaerobic environments, namely avoidance of anaerobiosis on the whole plant level, termed 'apparent' tolerance, and metabolic adaptation at the cellular and molecular levels, termed 'true' tolerance; (4) the importance of protein synthesis during hypoxia and anoxia for enhanced energy production and metabolic adaptation; (5) a general adaptive syndrome in plants to stress at the ultrastructural level and a possible molecular mechanism for its realization under anoxia; (6) the physiological role of anaerobically synthesized lipids and nitrate as alternative electron acceptors in an oxygen-free medium; and (7) the selection of cell lines derived from callus cultures that possess enhanced tolerance to anoxia and can regenerate whole plants with improved tolerance of soil waterlogging.

Int J Syst Evol Microbiol, 2002 Nov, 52(Pt 6), 2315 - 9
Citeromyces siamensis sp . nov., a novel halotolerant yeast isolated in Thailand; Nagatsuka Y et al.; Two halotolerant yeast strains, H130(T) and H149, were isolated from dry salted squid and fermented soybeans, respectively, in Thailand . Both isolates grew by multilateral budding, produced asci that had one roughened spherical ascospore and contained ubiquinone Q-8 . These characteristics were shared by Citeromyces matritensis, the only species of the genus Citeromyces . Strains H130(T) and H149 were differentiated from C matritensis by their ability to assimilate L-sorbose and L-lysine and to grow at 37 degrees C . The novel isolates were more tolerant to higher concentrations of cations (3 M NaCl or 0.8 M LiCI) and to higher osmotic pressure (60% glucose) than C . matritensis . A phylogenetic analysis of 18S rRNA gene sequence data indicated that the two novel isolates represented a sister species to C . matritensis . Furthermore, DNA-DNA hybridization data indicated that the isolates were clearly distinct from the type strain of C . matritensis (IFO 0954(T) . Based on the above characteristics, strains H130(T) and H149 are proposed to represent a novel species within the genus Citeromyces, Citeromyces siamensis; the type strain is H130(T) (= IFO 11052(T) = JCM 11522(T) = TISTR 5777(T) = CBS 9153(T)).

Pediatr Res, 2003 Jan, 53(1), 113 - 8
Nicotinic induction of preproenkephalin and tyrosine hydroxylase gene expression in butyrate-differentiated rat PC12 cells: a model for adaptation to gut-derived environmental signals; Nankova BB et al.; Accelerated maturation of peripheral sympathoadrenal transmitter levels and function occurs at 7-10 postnatal days in the rat . This event is temporally disconnected from the timing of major changes in physiologic stimuli evident after the birthing process (i.e . temperature, oxygen, sound, light, etc.) . Colonization of the gut, fermentation of carbohydrates, and production of short-chain fatty acids (e.g . butyrate) mirrors this postnatal time course . In this report, we examined the interaction between butyrate differentiation of rat pheochromocytoma cells and cholinergic-nicotinic induction of the neuropeptide (enkephalin) and catecholamine-related biosynthetic enzymes (tyrosine hydroxylase, dopamine beta-hydroxylase, phenylethanolamine N-methyltransferase) . Our results show that butyrate induces both preproenkephalin and tyrosine hydroxylase mRNA through a proximal promoter region and that this regulatory step is time and dose dependent . Moreover, there is an additional interaction with cholinergic-nicotinic inducible mechanisms consistent with classically described transsynaptic cholinergic regulation of these genes . Dopamine beta-hydroxylase and phenylethanolamine N-methyltransferase promoters were not affected by butyrate treatment . We speculate that colonization of the human gut (along with the attendant fermentation of enteral carbohydrates to short-chain fatty acids) may represent a mechanism through which environmental signals affect postnatal maturation of sympathoadrenal transmitter systems.

J Exp Bot, 2003 Jan, 54(382), 467 - 75
Metabolic signalling and carbon partitioning: role of Snf1-related (SnRK1) protein kinase; Halford NG et al.; A protein kinase that plays a key role in the global control of plant carbon metabolism is SnRK1 (sucrose non-fermenting-1-related protein kinase 1), so-called because of its homology and functional similarity with sucrose non-fermenting 1 (SNF1) of yeast . This article reviews studies on the characterization of SnRK1 gene families, SnRK1 regulation and function, interacting proteins, and the effects of manipulating SnRK1 activity on carbon metabolism and development.

Bioresour Technol, 2003 May, 87(3), 337 - 9
Grape and apple wines volatile fermentation products and possible relation to spoilage; Polychroniadou E et al.; The main volatile by-products of the alcoholic fermentation of grape wine, cider and apple pulp wine were investigated to determine if any correlated with spoilage resistance in the latter two . Spoilage was visually detected after seven days in low-alcohol grape wine in comparison to 11 and 16 days in cider and apple pulp wine, respectively . Acetaldehyde, ethyl acetate, methanol, propanol, isobutanol and amyl alcohols were the main fermentation by-products detected in all three wines . There were highest concentrations of acetaldehyde, ethyl acetate, methanol and propanol in grape wine and, therefore, these by-products could not be implicated in spoilage resistance in apple wines . Increased concentrations of isobutanol and amyl alcohols, however, in cider and apple pulp wine in comparison to grape wine might have been the reason for spoilage resistance in the apple wines.

Folia Microbiol (Praha), 2002, 47(5), 511 - 5
Influence of cultivation conditions on citrate production by Aspergillus niger in a semi-pilot-scale plant; Ul-Haq I et al.; The influence of some fermentation parameters on the semi-pilot scale (alteration of growth conditions, e.g., sugar concentration, incubation temperature and initial pH) on citrate production was demonstrated in parent and mutant strains of Aspergillus niger . Raw material from sugar industry (cane molasses) was examined as basal fermentation medium in a stirred stainless-steel 15-L fermentor . After growth on medium with 150 g/L sugar, the parent strain produced 51.2 g/L citric acid; the mutant strain achieved production maximum of 96.2 g/L . Comparing the growth, kinetic (volumetric substrate uptake rate, rate of substrate consumption and volumetric productivity rate) and production parameters it was found that the mutant strain grows more rapidly, with slightly changed morphology (intermediate, shiny round pellets with diameter 0.6-0.7 mm), and exhibits a higher citrate production and higher efficiency of sugar utilization.

Folia Microbiol (Praha), 2002, 47(5), 493 - 8
Isolation and identification of Streptomyces sp . and assay of its exocellular water-soluble blue pigments; Lu L et al.; A bacterial strain producing a great amount of blue pigment during submerse fermentation was isolated and identified . Based on morphological characteristics, cell-wall chemotype and sequence of 16S rRNA gene, the strain should belong to the genus Streptomyces; it had 99.4% homology of 16S rRNA gene sequence with that of Streptomyces indigocolor . The pigment production by the strain was affected by carbon and nitrogen sources . The main components of the pigment mixture (detected by HPLC and TLC) were tentatively classified as actinorhodin-related compounds . The pigment was relatively stable against light and higher temperature but was sensitive to low pH . The preliminary acute-toxicity determination showed that the pigment was nontoxic (LD50 > 15 mg/g).

Biotechniques, 2002 Dec, 33(6), 1296 - 300
Buffered non-fermenter system for lab-scale production of secreted recombinant His-tagged proteins in Saccharomyces cerevisiae; Ngamkitidechakul C et al.; Expression of recombinant proteins using a secretion system can minimize co-purification of contaminating host proteins . Production of His-tagged recombinant proteins in the yeast alpha-factor secretion system has previously required a fermenter system to control the growth conditions such as pH of the yeast culture . We describe an inexpensive non-fermenter system for the production of secreted recombinant His-tagged proteins in Saccharomyces cerevisiae that uses a buffered low peptone YP glycerol medium, which does not interfere with immobilized metal affinity chromatography . Maspin, a tumor suppressor serpin, was expressed as a secreted N-terminal His/FLAG-tagged protein . Purification of the soluble active recombinant protein only requires centrifugation, concentration by ultrafiltration, and Ni2+ affinity chromatography . Purified protein yields of this system are 3-5 mg/L culture medium.

Microbiol Res, 2002, 157(4), 267 - 74
Isolation and identification of yeasts associated with vineyard and winery by RFLP analysis of ribosomal genes and mitochondrial DNA; Sabate J et al.; Yeast colonies isolated from vineyard and cellar substrates were analysed in the present study . Yeast species assessment was carried out by amplification and digestion of a region of the ribosomal RNA gene repeat unit . Saccharomyces strains were also characterised using mitochondrial DNA restriction analysis . Oxidative basidiomycetous yeasts without enological potential were predominant in the vineyard environment . Yeasts associated with grape skin depend on grape variety, vintage and degree of grape maturation . These species from grape surface constituted the predominant microbiota in must and they developed during the first stages of the process . Yeasts colonies were also isolated and identified from the walls of a fermentation vat some days before the harvest . Contray to what was expected, Saccharomyces cerevisiae was not the major species isolated as Candida sorbosa represented 76% of the species isolated . Saccharomyces strains isolated from the fermentation vat had been previously isolated in wine fermentations in this cellar . Therefore, these strains should be considered as constant residents of this winery.

J Gen Appl Microbiol, 2002 Oct, 48(5), 261 - 7
Isolation and clonal pre-selection of enological Saccharomyces; Caridi A et al.; The aim of the present study was to perform a fast pre-selection from a great number of wine yeasts using a simple phenotypic-based methodology that allows many different strains to be simultaneously tested . A total of 150 elliptic yeasts, isolated from must and wine from black grapes of a distinctive Italian variety, were studied . Yeasts were identified to genus level by assessing their ability to ferment glucose and their production of spores on acetate agar . The Saccharomyces strains were seeded on BiGGY agar to determine their H(2)S production, on calcium carbonate agar to test their acetic acid production, and on grape-skin agar and on grape-seed agar to assess their interaction with phenolic compounds . The Saccharomyces strains were also examined for fermentative vigor after 2 d or 7 d both with and without the addition of 100 mg L(-1) of SO(2) in must at 20 degrees brix and pH 3.20 . At the end of fermentation, the wines produced by the 18 best yeasts were analyzed and the strains were studied for additional biochemical and technological characteristics . The resistance of the strains to simultaneous acid-stress and osmotic-stress was studied carrying out in duplicate winemaking tests in must at 30 degrees brix and pH 2.60 . A remarkable heterogeneity among the 150 autochthonous yeasts studied was demonstrated . The phenotypical biodiversity is particularly interesting for several technological characteristics useful in winemaking, such as fermentation vigor, acetic acid production and malic acid content of the wines . The vast majority of the elliptic wine yeasts isolated did not show suitable characteristics, so only 18 strains, 12% of the total, remained for the final tests . Many of the strains that had passed the preliminary screenings revealed some defects when they were studied for fermentation performance, both in standard winemaking and under stressors . Two strains exhibited particularly interesting performances: one strain for winemaking of normal musts and the other for winemaking of musts from dried grapes or under stressful conditions.

J Gen Appl Microbiol, 1998 Oct, 44(5), 347 - 353
Breeding of flocculent industrial alcohol yeast strains by self-cloning of the flocculation gene FLO1 and repeated-batch fermentation by transformants; Ishida-Fujii K et al.; A nonflocculent industrial polyploid yeast strain, Saccharomyces cerevisiae 396-9-6V, was converted to a flocculent one by introducing a functional FLO1 gene at the URA3 locus . The flocculent strain FSC27 obtained was a so-called self-cloned strain, having no bacterial DNA . FSC27 cells could be easily recovered for reuse from fermentation mash without any physical energy . The strain produced a concentration of alcohol as high as 396-9-6V, although the fermentation rate of FSC27 was slightly lower than that of 396-9-6V . When uracil was added to the medium or when URA3 was reintroduced into FSC27 (named FSCU-L18), the fermentation rate and the growth rate increased, and the ethanol concentration produced was higher than that produced by the parent strain . The stable flocculation and high ethanol productivity were observed by using FSCU-L18 during 10 cycles of repeated-batch fermentation test.

J Gen Appl Microbiol, 1999 Feb, 45(1), 23 - 28
Identification of acetic acid bacteria isolated from Indonesian sources, especially of isolates classified in the genus Gluconobacter; Yamada Y et al.; Sixty-four strains of acetic acid bacteria were isolated from Indonesian sources such as fruits, flowers, and fermented foods by the enrichment culture at pH 3.5 . Forty-five strains were routinely identified as Acetobacter strains because of their oxidation of acetate and lactate to carbon dioxide and water and their Q-9 isoprenolog, corresponding to 70% of all the 64 acetic acid bacteria isolated . Eight isolates were identified as Gluconacetobacter strains because of their oxidation of acetate and lactate and their Q-10 isoprenolog, occupying 13% of all the isolates . The remaining 11 isolates, accommodated in the genus Gluconobacter because of no oxidation of acetate and lactate and because of their Q-10 isoprenolog, accounted for 17% of all the isolates . They were divided into two groups based on DNA base compositions . One comprised the seven isolates, which had high G1C contents of DNA ranging from 60.3 to 63.5 mol% and of which DNAs hybridized with that of the type strain of Gluconobacter oxydans at values of 64-94% of DNA relatedness . The other comprised the remaining four isolates, which had low G+C contents of DNA ranging from 57.5 to 57.7 mol% and of which DNAs hybridized with that of the type strain of Gluconobacter frateurii at values of 63-77% of DNA relatedness . The high values of DNA relatedness, 84 to 96%, were obtained between the type strains of Gluconobacter cerinus and Gluconobacter asaii.

J Gen Appl Microbiol, 1999 Feb, 45(1), 1 - 6
Isolation and characterization of triacylglycerol-secreting mutant strain from yeast, Saccharomyces cerevisiae; Nojima Y et al.; To establish the molecular bases for development of a microbiological system approaching excretive fermentation of useful lipids, a mutant strain that accumulates lipids in the medium was isolated from the laboratory yeast Saccharomyces cerevisiae . Following the mutagenesis to strain YP1, a long chain fatty acid utilizer with ethylmethane sulfonate, the mutant strain, STG1, was selected from about 80,000 colonies . The analysis of extracellular lipids and the monitoring of leakage of intracellular proteins indicated that strain STG1 secreted lipids containing triacylglycerols into the extracellular space without cell lysis . Genetic studies clarified that this mutation was recessive and was complemented by wild-type genomic DNA fragments . STG1 was considered to be a good tool for elucidation of the molecular mechanism for transmembrane lipid transport.

J Gen Appl Microbiol, 1999 Jun, 45(3), 99 - 103
Production of alkaline protease by a genetically engineered Aspergillus oryzae U1521; Samarntarn W et al.; The production of alkaline protease of Aspergillus oryzae U1521 was examined in liquid culture . In a culture of defatted soybean only, it gave satisfactory enzyme yields at 584,000 U/g defatted soybean . When various carbohydrates were supplemented, enzyme production was significantly increased . An increase in production by lactose was the most marked . Enrichment with casitone or casein increased productivity, but not cornsteep solid . Media formulation (g/L) of defatted soybean 10, lactose 5, casitone 1, and KH(2)PO(4) 5 enhanced alkaline protease production by A . oryzae U1521 to a maximum of 1,410,000 U/g defatted soybean . Scaling-up experiments indicated the flask-scale results could be reproduced at 40 g of substrate in 5-L fermenter . The enzyme activity was maximum between pH 8-9 and at a temperature of 45 degrees C.

J Gen Appl Microbiol, 1999 Dec, 45(6), 277 - 282
Effect of three strains of Pleurotus tuber-regium (Fr.) Sing . on chemical composition and rumen fermentation of wheat straw; Jalc D et al.; This study was conducted to investigate changes in in vitro dry matter digestibility (IVDMD) and cell wall constituent degradation in wheat straw treated with 3 strains of the fungus Pleurotus tuber-regium (PT) . The incubation of wheat straw for 30 days at 28 degrees C improved IVDMD from 30.3% (UWS-untreated wheat straw) to 47.1% for strain PT1, to 48.5% for PT4, and was unchanged IVDMD-29.9% -for PT5 . The growth of fungi was accompanied by the dry matter loss of wheat straw: 31.5% for PT1, 20.9% for PT4, and 4.8% for PT5 . Fungal treatment was characterized by increased crude protein and ash contents (%) in all fungi-treated straws and reduced hemicellulose and lignin content . It is evident that enzymes of all 3 PT strains preferentially degraded hemicellulose and lignin over cellulose . Wheat straw treated with PT1 (TWS-PT1), PT4 (TWS-PT4), and PT5 (TWS-PT5) and barley (80% : 20%) were used as the experimental diets at the fermentation in the artificial rumen . UWS with barley (80% : 20%) served as the control diet . The fermentation of experimental diets was accompanied with increased IVDMD and a very low degree of hemicellulose degradation . Total gas and methane productions were similar in all diets . Moreover, total volatile fatty acid (VFA) production (mmol day(-1)), mol % of acetate, propionate, butyrate, isobutyrate, and isovalerate were not influenced during the fermentation of experimental diets . From the stoichiometric relations, production, utilization, and recovery of metabolic hydrogen and organic matter fermented were unchanged . Only the recovery of metabolic hydrogen in TWS-PT5 was significantly increased in comparison to control diet . Total microbial production showed the tendency of lower values in experimental diets, and it was accompanied with a significant decrease of ammonia nitrogen (mg L(-1)) . Finally the results showed that the strains of Pleurotus tuber-regium can improve the quality of wheat straw, but the loss of dry matter (DM) (mainly hemicellulose) limits the effective utilization of fungi-treated straw in ruminant digestion.

J Gen Appl Microbiol, 1997 Jun, 43(3), 133 - 137
Effect of six species of white-rot basidiomycetes on the chemical composition and rumen degradability of wheat straw; Jalc D et al.; This study was conducted to investigate changes in in vitro dry matter digestibility (IVDMD), volatile fatty acids (VFA) production and cell-wall constituent degradation in wheat straw treated with six white-rot fungi: Daedalea quercina, Hericium clathroides, Phelinus laevigatus, Inonotus andersonii, Inonotus obliquus, and Inonotus dryophilus . The incubation of wheat straw for 30 days at 28 C improved IVDMD from 41.4 (control) to 59.2% for D . quercina, 56.3% for H . clathroides, 50.2% for P . laevigatus, 51.4% for I . andersonii, 52% for I . obliquus, and 55.9% for I . dryophilus . In contrast, the growth of fungi was accompanied by the dry matter loss of wheat straw: 43% for D . quercina, 12% for H . clathroides, and 22-25% for the other fungi . It is evident that the increase in digestibility by D . quercina was not offset by a loss of dry matter . The total VFA production during the rumen fermentation of fungus-treated straw was slightly increased by H . clathroides and I . dryophilus only . Neutral detergent fiber (NDF) and acid detergent fiber (ADF) were reduced in fungus-treated straw . Out of the three fractions (hemicellulose, cellulose, and lignin), hemicellulose and lignin showed the largest proportionate loss after inoculation with the fungi D . quercina, H . clathroides, P . laevigatus, and I . obliquus . The other two fungi showed the largest proportionate loss in cellulose and hemicellulose contents . The results of this study suggest that the digestion enhancement of wheat straw colonized by white-rot fungi is regulated by complex factors including the degradation of structural carbohydrates and lignin.

Biofizika, 2002 Nov-Dec, 47(6), 1064 - 7
{Interaction of membrane proton conductivity, membrane and oxidation-reduction potential in Escherichia coli}; Akopian K et al.; It was shown that the proton conductivity of Escherichia coli membranes depends on pH and other conditions of bacterial growth . It is considerably lower in cells fermenting glucose and accomplishing the nitrate-nitrite respiration compared with cells accomplishing the oxygen respiration . Proton conductivity increases substantially with decreasing pH of medium . It was found that proton conductivity is related to the redox and membrane potentials of cells . The energy-dependent flux of protons from cells and the ATPase activity of membrane vesicles considerably vary depending on whether bacteria are grown under aerobic or anaerobic conditions . The H+ flux from cells fermenting glucose (pH 7.5) was 1.7 times greater than the H+ flux from cells that accomplish the nitrate-nitrite and oxygen respiration . The N,N'-dicyclohexylcarbodiimide (DCCD)-sensitive ATPase activity increased 2.5 times as K+ concentration increased to 100 mM (including residual K+ in potassium-free medium) . The DCCD-sensitive ATPase activity considerably decreased with decreasing pH of medium, whereas the ATPase activity that was not suppressed by DCCD was stimulated . These results can be used for establishing the relationship between membrane proton conductivity and the energy-dependent H+ flux and ATPase activity.

Br J Nutr, 2002 Dec, 88 Suppl 3, S281 - 5
Bioavailability of minerals in legumes; Sandberg AS; The mineral content of legumes is generally high, but the bioavailability is poor due to the presence of phytate, which is a main inhibitor of Fe and Zn absorption . Some legumes also contain considerable amounts of Fe-binding polyphenols inhibiting Fe absorption . Furthermore, soya protein per se has an inhibiting effect on Fe absorption . Efficient removal of phytate, and probably also polyphenols, can be obtained by enzymatic degradation during food processing, either by increasing the activity of the naturally occurring plant phytases and polyphenol degrading enzymes, or by addition of enzyme preparations . Biological food processing techniques that increase the activity of the native enzymes are soaking, germination, hydrothermal treatment and fermentation . Food processing can be optimized towards highest phytate degradation provided that the optimal conditions for phytase activity in the plant is known . In contrast to cereals, some legumes have highest phytate degradation at neutral or alkaline pH . Addition of microbial enzyme preparations seems to be the most efficient for complete degradation during processing . Fe and Zn absorption have been shown to be low from legume-based diets . It has also been demonstrated that nutritional Fe deficiency reaches its greatest prevalence in populations subsisting on cereal- and legume-based diets . However, in a balanced diet containing animal protein a high intake of legumes is not considered a risk in terms of mineral supply . Furthermore, once phytate, and in certain legumes polyphenols, is degraded, legumes would become good sources of Fe and Zn as the content of these minerals is high.

Appl Biochem Biotechnol, 2003 Jan, 104(1), 23 - 36
Effect of freeze-dried immobilized cells on delignified cellulosic material in low-temperature and ambient-temperature wine making; Ikonomopoulou M et al.; In this article, we report on wine making by freeze-dried immobilized cells on delignified cellulosic material for ambient and low temperatures . Biocatalyst supported by freeze-dried delignified cellulosic (FDC) material recovered after the first repeated-batch fermentations the fermentation efficiency and startup, which become about equal to those of biocatalyst supported by wet delignified cellulosic material . The FDC biocatalyst was suitable for wine making at low temperatures (5-15 degrees C), and produced wine of 12% alcoholic degree, with the main volatiles contained in the wine and reduced by a decrease in temperature . The fermentation efficiency was not affected by total acidity of must, while an increase in initial Be density improved percentages of higher alcohols and ethyl acetate . The quality of the wine was validated by a preliminary taste test to be in the range of acceptable to excellent.

J Food Prot, 2002 Dec, 65(12), 1854 - 60
Fate of Escherichia coli O157:H7 during silage fermentation; Byrne CM et al.; The survival characteristics of Escherichia coli O157:H7 in silage derived from contaminated grass were investigated . The survival of other enteric bacteria was also investigated to determine if E . coli O157:H7 demonstrates enhanced acid tolerance in comparison . Samples of chopped grass were treated as follows: (i) no additive (control); (ii) inoculation with E . coli O157:H7 to a final concentration of log10 4.0 CFU g(-1); (iii) addition of an 85% solution of formic acid at 3.0 ml kg(-1) grass; and (iv) addition of both E . coli O157:H7 and formic acid, at the above concentrations . Treated 6-kg grass samples were packed into laboratory silos, sealed, and stored at 15 degrees C for up to 180 days . Individual replicate silos were removed from storage periodically and subjected to microbiological and chemical analyses . Chemical analyses of the silage samples indicated that lactic acid-dominant fermentations, with a rapid drop in pH, occurred . Numbers of enteric bacteria decreased from log10 7.0 to 8.0 CFU g(-1) to undetectable levels within 19 days' storage . E . coli O157:H7 did not survive the silage fermentation process, with numbers declining from approximately log10 4.0 CFU g(-1) to undetectable levels within 19 days of ensiling . The pattern of decline in numbers of E . coli O157:H7 was the same as that for the enteric bacteria, indicating that under the conditions tested, the acid tolerance of E . coli O157:H7 was not significantly different from the acid tolerance of other enteric bacteria . This study found that E . coli O157:H7 did not survive a good silage fermentation process, indicating that properly ensiled grass that is correctly stored is unlikely to be a vector for the transmission of the pathogen among cattle.

J Appl Microbiol, 2003, 94(1), 120 - 6
Optimization of submerged culture process for the production of mycelial biomass and exo-polysaccharides by Cordyceps militaris C738; Kim SW et al.; AIMS: The objective of the present study was to determine the optimal culture conditions for mycelial biomass and exo-polysaccharide (EPS) by Cordyceps militaris C738 in submerged culture . METHODS AND RESULTS: The optimal temperatures for mycelial biomass and EPS production were 20 degrees C and 25 degrees C, respectively, and corresponding optimal initial pHs were found to be 9 and 6, respectively . The suggested medium composition for EPS production was as follows: 6% (w/v) sucrose, 1% (w/v) polypeptone, and 0.05% (w/v) K2HPO4 . The influence of pH on the fermentation broth rheology, morphology and EPS production of C . militaris C738 was carried out in a 5-l stirred-tank fermenter . The morphological properties were comparatively characterized by pellet roughness and compactness by use of image analyser between the culture conditions with and without pH control . The roughness and compactness of the pellets indicated higher values at pH-stat culture (pH 6.0), suggesting that larger and more compact pellets were desirable for polysaccharide production (0.91 g g(-1) cell d(-1) . CONCLUSIONS: Under the optimized culture conditions (with pH control at 6), the maximum concentration of biomass and EPS were 12.7 g l(-1) and 7.3 g l(-1), respectively, in a 5-l stirred-tank fermenter . SIGNIFICANCE AND IMPACT OF THE STUDY: The critical effect of pH on fungal morphology and rheology presented in this study can be widely applied to other mushroom fermentation processes.

Biotechnol Bioeng, 2003 Feb 20, 81(4), 448 - 58
Determination of in vivo oxygen uptake and carbon dioxide evolution rates from off-gas measurements under highly dynamic conditions; Wu L et al.; In vivo kinetics of Saccharomyces cerevisiae are studied, in a time window of 150 s, by analyzing the response of O(2) and CO(2) in the fermentor off-gas after perturbation of chemostat cultures by metabolite pulses . Here, a new mathematical method is presented for the estimation of the in vivo oxygen uptake rate (OUR) and carbon dioxide evolution rate (CER) directly from the off-gas data in such perturbation experiments . The mathematical construction allows effective elimination of delay and distortion in the off-gas measurement signal under highly dynamic conditions . A black box model for the fermentor off-gas system is first obtained by system identification, followed by the construction of an optimal linear filter, based on the identified off-gas model . The method is applied to glucose and ethanol pulses performed on chemostat cultures of S . cerevisiae . The estimated OUR is shown to be consistent with the independent dissolved oxygen measurement . The estimated in vivo OUR and CER provide valuable insights into the complex dynamic behavior of yeast and are essential for the establishment and validation of in vivo kinetic models of primary metabolism .

Biotechnol Bioeng, 2003 Feb 20, 81(4), 438 - 47
Use of confocal microscopy to follow the development of penetrative hyphae during growth of Rhizopus oligosporus in an artificial solid-state fermentation system; Nopharatana M et al.; Two methods were compared for determining the concentration of penetrative biomass during growth of Rhizopus oligosporus on an artificial solid substrate consisting of an inert gel and starch as the sole source of carbon and energy . The first method was based on the use of a hand microtome to make sections of approximately 0.2- to 0.4-mm thickness parallel to the substrate surface and the determination of the glucosamine content in each slice . Use of glucosamine measurements to estimate biomass concentrations was shown to be problematic due to the large variations in glucosamine content with mycelial age . The second method was a novel method based on the use of confocal scanning laser microscopy to estimate the fractional volume occupied by the biomass . Although it is not simple to translate fractional volumes into dry weights of hyphae due to the lack of experimentally determined conversion factors, measurement of the fractional volumes in themselves is useful for characterizing fungal penetration into the substrate . Growth of penetrative biomass in the artificial model substrate showed two forms of growth with an indistinct mass in the region close to the substrate surface and a few hyphae penetrating perpendicularly to the surface in regions further away from the substrate surface . The biomass profiles against depth obtained from the confocal microscopy showed two linear regions on log-linear plots, which are possibly related to different oxygen availability at different depths within the substrate . Confocal microscopy has the potential to be a powerful tool in the investigation of fungal growth mechanisms in solid-state fermentation .

Curr Genet, 2002 Dec, 42(3), 147 - 52 Epub 2002 Nov 29.
Up-regulation of tRNA biosynthesis affects translational readthrough in maf1-delta mutant of Saccharomyces cerevisiae; Kwapisz M et al.; Maf1p is a negative effector of RNA polymerase III in yeast . The maf1-delta mutation caused an increase in the level of cellular tRNAs, but a decrease of translational readthrough at nonsense codons . Using the lacZ- luc dual gene reporter system, we detected an almost twofold diminution of UAA and UAG readthrough in maf1-delta compared with the parental strain . The maf1-delta mutation did not affect the rate of protein biosynthesis and growth at standard conditions, but resulted in temperature-sensitive growth on non-fermentable carbon sources . We examined the correlation of the temperature sensitive and antisuppression phenotypes of maf1- Delta using a colour phenotype assay in the ade2-1 SUP11 strain . Antisuppression, but not the temperature-sensitive growth defect, was compensated either by increased dosage of SUP11or by {PSI(+)}, the prion form of the translation termination factor Sup35p . Summarizing, the elevated tRNA levels in maf1- Delta increase translational fidelity and, independently, affect growth under special conditions.

J Dairy Sci, 2002 Nov, 85(11), 2964 - 73
Ruminal digestion and fermentation of high-producing dairy cows with three different feeding systems combining pasture and total mixed rations; Bargo F et al.; Six multiparous Holstein cows fitted with rumen cannulas were used to study the effect of three feeding systems combining pasture and total mixed rations (TMR) on ruminal digestion in a 21-wk repeated measures experiment . The three treatments were: 1) pasture plus concentrate (PC), 2) pasture plus partial TMR (pTMR), and 3) TMR (nonpasture) . Ruminal NH3-N concentration was lower on both the pTMR and TMR treatments (10.2 +/- 0.5 mg/dL) than on the PC treatment (19.9 +/- 0.5 mg/dL) . Ruminal pH was not affected by treatments and averaged 5.87 . Neither total volatile fatty acid concentration (137.5 mmol/L) nor individual volatile fatty acid proportions (63.1,20.6, and 12.0 mol/ 100 mol for acetate, propionate, and butyrate, respectively) differed among treatments . The pTMR treatment reduced the total potentially degradable fraction of dry matter (85.5 vs . 82.3%) and the potentially digestible fraction of neutral detergent fiber (82.1 vs . 74.9%) of pasture compared to the PC treatment . Ruminal NH3-N losses were reduced when combining pasture and TMR; however this combination decreased the ruminal digestion of pasture, indicating the presence of associative effects in the rumen.

J Dairy Sci, 2002 Nov, 85(11), 2928 - 47
Corn silage management III: effects of hybrid, maturity, and processing on nitrogen metabolism and ruminal fermentation; Johnson LM et al.; Two experiments were conducted to evaluate the effects of maturity and mechanical processing of two hybrids of whole plant corn silage on DM and OM digestibility, nitrogen metabolism, ruminal fermentation, and milk production and composition in lactating Holstein cows . In the first experiment, Pioneer hybrid 3845 whole plant corn was harvested at hard dough, one-third milkline, and two-thirds milkline with a theoretical length-of-cut of 6.4 mm . At each stage of maturity, corn was harvested with (1-mm roll clearance) and without (15.9-mm roll clearance) mechanical processing using a John Deere 5830 harvester with an on-board kernel processor . In the second experiment, Pioneer hybrids 3845 and Quanta were harvested at one-third milkline, two-thirds milkline, and blackline stages of maturity with and without mechanical processing . The theoretical length-of-cut was 12.7 mm . Total tract DM and OM digestibilities were lower for cows fed diets containing processed corn silage in experiment 1, and tended to be lower for cows fed diets containing unprocessed corn silage in experiment 2 . Ruminal acetate concentrations were greater and ruminal propionate concentrations were lower 2 and 6 h after feeding for cows fed diets containing corn silage harvested at physiological maturity in experiment 2 . This was due to decreased digestion of starch at advanced maturities in experiment 2 . Ruminal pH tended to decline rapidly after feeding for cows fed hybrid Quanta (2 h) compared to hybrid 3845 (5 h) corn silage based diets . Ruminal acetate concentrations decreased and ruminal propionate concentrations increased 2 and 6 h after feeding for cows fed diets containing hybrid Quanta corn silage compared to hybrid 3845 corn silage . This was related to a greater starch concentration in the corn silage, greater starch intake, and increased rate of starch digestion for cows fed hybrid Quanta corn silage-based diets . Microbial nitrogen flow was lower and feed nitrogen flow was greater for cows fed diets containing hybrid Quanta corn silage . The lower microbial nitrogen flow was due to lower microbial nitrogen concentration and nonammonia nitrogen flow to the duodenum . Milk fat and protein concentrations had a strong quadratic relationship with forage NDF intake as a percentage of body weight . When forage NDF intake as a percentage of body weight dropped below 0.70%, there was a rapid decline in milk fat and protein concentrations.

J Dairy Sci, 2002 Nov, 85(11), 2890 - 904
Ruminal fermentation and nutrient digestion by dairy cows fed varying amounts of soyhulls as a replacement for corn grain; Ipharraguerre IR et al.; Five multiparous Holstein cows cannulated in the rumen and duodenum that averaged 63 d in milk were used in a 5 x 5 Latin square design with 14-d periods to evaluate the incremental substitution of soyhulls for corn in the diet . Diets contained 23% alfalfa silage, 23% corn silage, and 54% concentrate on a dry matter (DM) basis . Pelleted soyhulls replaced corn in the concentrate to supply 0, 10, 20, 30, or 40% of the dietary DM . The intakes of DM and organic matter were unaffected by treatments . Intakes of acid detergent fiber and neutral detergent fiber increased linearly, but the intake of nonstructural carbohydrates decreased linearly as soyhulls increased from 0 to 40% of dietary DM . The amount of acid detergent fiber and neutral detergent fiber digested was increased whereas the amount of nonstructural carbohydrate digested was decreased in the rumen, in the lower digestive tract, and in the total digestive tract as soyhulls replaced corn in the diet . Passage to the duodenum of nonammonia N, microbial N, nonammonia nonmicrobial N, total essential amino acids, total nonessential amino acids, and total amino acids were not affected by treatments . Yield of milk (29.5 kg/d) was not affected by treatments in this experiment . In a companion experiment, cows fed the 40% SH diet produced 1.2 kg/day per cow less (P < 0.07) milk than cows fed the control diet which is similar to the 1.3 kg/day per cow less milk produced by cows fed the same 40% SH diet in this experiment . Differences in the source of energy (fiber vs . nonstructural carbohydrates), in the amount of fiber and nonstructural carbohydrates digested, and in the site of digestion in the gastrointestinal tract may cause a shortage of the source and/or amount of energy that is required for maximum milk production in high producing cows when more than 30% of the dietary DM that is supplied as corn is replaced with soyhullss.

Crit Rev Biotechnol, 2002, 22(4), 355 - 74
Production and mass transfer characteristics of non-Newtonian biopolymers for biomedical applications; Richard A et al.; The market for microbial biopolymers is currently expanding to include several emerging biomedical applications . Specifically, these applications are drug delivery and wound healing . A fundamental understanding of the key fermentation parameters is necessary in order to optimize the production of these biopolymers . Considering that most microbial biopolymer systems exhibit non-Newtonian rheology, oxygen mass transfer can be an important parameter to optimize and control . In this article, we present a critical review of recent advances in rheological and mass transfer characteristics of selected biopolymers of commercial interest in biomedical applications.

Exp Biol Med (Maywood), 2002 Dec, 227(11), 1017 - 21
Modulation of CD8+ intraepithelial lymphocyte distribution by dietary fiber in the rat large intestine; Ishizuka S et al.; We studied whether ingestion of dietary fiber modifies the distribution of intraepithelial lymphocytes (IEL) in a physiological condition . Male WKAH rats were fed diets either with fiber (sugar beet fiber or crystalline cellulose, 100 g/kg diet each) or without fiber for 3 weeks . The number of CD8(+), CD4(+), and NKR-P1(+) IEL per epithelial layer in the crypt section of the cecum, proximal colon, and distal colon were scored by immunohistochemical staining . We found that the proportion of CD8(+) IEL was greater in the cecal mucosa and was gradually reduced toward the distal large intestine in general . In contrast, there was no difference in the proportion of CD4(+) and NKR-P1(+) IEL in the large intestine . Dietary sugar beet fiber, but not crystalline cellulose, increased the proportion of CD8(+) IEL, especially in the cecal mucosa, but not the CD4(+) and NKR-P1(+) IEL . Analysis of cecal organic acid concentration confirmed higher concentrations of acetate and butyrate, and lower concentration of succinate and isovalerate, in the cecum of the rats fed sugar beet fiber than other diets . These results indicate that ingestion of some dietary fiber modulates local cell proliferation of a progenitor of CD8(+) IEL or promotes homing of CD8(+) T cells into the large intestinal epithelium, most likely via the fermentation in the luminal contents.

Lett Appl Microbiol, 2003, 36(1), 54 - 8
Saccharomyces cerevisiae biodiversity in spontaneous commercial fermentations of grape musts with 'adequate' and 'inadequate' assimilable-nitrogen content; Granchi L et al.; AIM: To evaluate whether intraspecific diversity of Saccharomyces cerevisiae in wine fermentations is affected by initial assimilable-nitrogen content . METHODS AND RESULTS: Saccharomyces cerevisiae isolates from two spontaneous commercial wine fermentations started with adequate and inadequate nitrogen amounts were characterized by mitochondrial DNA restriction analysis . Several strains occurred in each fermentation, two strains, but not the same ones, being predominant at frequencies of about 30% . No significant differences were detected by comparing the biodiversity indices of the two fermentations . Cluster analysis demonstrated that the strain distribution was independent of nitrogen content, the two pairs of closely related dominant strains grouping into clusters at low similarity . CONCLUSIONS: The genetic variability of S . cerevisiae in wine fermentations seemed not to depend on the nitrogen availability in must . SIGNIFICANCE AND IMPACT OF THE STUDY: Nitrogen content did not affect the genetic diversity but may have induced a 'selection effect' on S . cerevisiae strains dominating wine fermentations, with possible consequences on wine properties.

J Gen Appl Microbiol, 2001 Jun, 47(3), 119 - 131
Identification of Acetobacter strains isolated from Indonesian sources, and proposals of Acetobacter syzygii sp . nov., Acetobacter cibinongensis sp . nov., and Acetobacter orientalis sp . nov .
Lisdiyanti P, Kawasaki H, Seki T, Yamada Y, Uchimura T, Komagata K.
Forty-six strains of acetic acid bacteria newly isolated from flowers, fruits, and fermented foods collected in Indonesia were taxonomically studied . They were Gram-negative rods, produced acetic acid from ethanol, oxidized acetate and lactate to CO(2) and H(2)O, and had Q-9 as the major ubiquinone system . On the basis of DNA-DNA similarity, all strains studied, including type strains and reference strains of the genus Acetobacter, were separated into eleven groups (Groups I to XI) . Of the 46 isolates, two isolates were included in Group II and identified as Acetobacter pasteurianus, five in Group IV as A . orleanensis, 16 in Group V as A . lovaniensis, five in Group VII as A . indonesiensis, and three in Group VIII as A . tropicalis . The remaining 15 isolates constituted three new groups based on DNA-DNA similarity; four isolates were included in Group IX, two in Group X, and nine in Group XI . No isolates were identified as A . aceti (Group I), A . peroxydans (Group III), and A . estunensis (Group VI) . Phylogenetic analysis based on 16S rDNA sequences of representative strains of the Groups indicated belonging to the strains of the genus Acetobacter . On the basis of DNA base composition, DNA-DNA similarity, and 16S rDNA sequences, three new species of the genus Acetobacter are proposed: Acetobacter syzygii sp . nov . for Group IX, Acetobacter cibinongensis sp . nov . for Group X, and Acetobacter orientalis sp . nov . for Group XI . The distribution of Acetobacter strains in Indonesia is discussed in light of isolation sources.

J Gen Appl Microbiol, 2001 Jun, 47(3), 107 - 117
Structure and function of cyclin-dependent Pho85 kinase of Saccharomyces cerevisiae; Toh-E A et al.; Yeast Saccharomyces cerevisiae has five cyclin-dependent protein kinases (Cdks), Cdc28, Srb10, Kin28, Ctk1, and Pho85 . Any of these Cdks requires a cyclin partner for its kinase activity and a Cdk/cyclin complex, thus produced, phosphorylates a set of specific substrate proteins to exert its function . The cyclin partners of Srb10, Kin28, and Ctk1 are Srb11, Ccl1, and Ctk2, respectively . In contrast to the fact that each of Srb10, Kin28, and Ctk1 has a single cyclin partner, Cdc28 and Pho85 are polygamous; Cdc28 has 9 cyclins and Pho85 has 10 cyclins . Among these Cdks, Kin28 and Cdc28 are essential Cdks and it is well known that Cdc28 kinase plays a major role in regulating cell cycle progression . Pho85 is a non-essential Cdk but its absence causes a broad spectrum of phenotypes such as constitutive expression of PHO5, inability to utilize non-fermentable carbon sources, defects in cell cycle progression, and so on . Pho85 homologues are expanding to higher eukaryotes . Pho85 is most closely related with Cdk5 in terms of the amino acid sequence . The functional analysis of the domains of Pho85 also supports the close relationship between Pho85 and Cdk5, in which it was shown that the method of regulation of these two kinases is similar . Furthermore, forced expression of the mammalian CDK5 gene in a pho85Delta strain canceled a part of the pho85 defects . In this review, we summarize the functions of both Pho85/cyclin kinase and emphasize yeast Pho85 as valuable model systems to elucidate the functions of their homologues in other organisms.

J Ind Microbiol Biotechnol, 2002 Dec, 29(6), 314 - 22
Influence of the yeast strain on the changes of the amino acids, peptides and proteins during sparkling wine production by the traditional method; Martinez-Rodriguez AJ et al.; The influence of five yeast strains on the nitrogen fractions, amino acids, peptides and proteins, during 12 months of aging of sparkling wines produced by the traditional or Champenoise method, was studied . High-performance liquid chromatography (HPLC) techniques were used for analysis of the amino acid and peptide fractions . Proteins plus polypeptides were determined by the colorimetric Bradford method . Four main stages were detected in the aging of wines with yeast . In the first stage, a second fermentation took place; amino acids and proteins plus polypeptides diminished, and peptides were liberated . In the second stage, there was a release of amino acids and proteins, and peptides were degraded . In the third stage, the release of proteins and peptides predominated . In the fourth stage, the amino acid concentration diminished . The yeast strain used influenced the content of free amino acids and peptides and the aging time in all the nitrogen fractions.

J Hepatobiliary Pancreat Surg, 2002, 9(4), 448 - 58
Enteral nutrition in HPB surgery: past and future; Bengmark S; Perioperative nutrition has, during the past century, been transformed from a tool to provide calorie and nitrogen support to a tool to boost the immune system and increase resistance to complications . Despite all the progress in medicine and surgery, perioperative morbidity, the rate of infections, thrombosis, and the development of serosal adhesions has remained the same as long as can be judged, or at least during the past 80 years . Most prone to develop complications are persons above the age of 65 and persons with depressed immunity . About 80% of the immune system is localized in the gastrointestinal tract, which offers great opportunities for modulation through enteral nutrition . As the stomach has a tendency to develop postoperative paralysis, tube feeding is often necessary . In 1918, Andresen demonstrated the advantages of enteral nutrition, which already started on the operating table . Mulholland and colleagues and Rhoads and co-workers demonstrated, during the 1940s, certain advantages of enteral tube feeding . Also, the works by Alexander, Fischer, and Ryan, and their co-workers supported the value of early enteral feeding, and suggested enteral feeding as an effective tool to boost the immune system . It was, however, works published in the early 1990s, by Moore and colleagues and by Kudsk and colleagues, which made surgeons more aware of the advantages of early enteral nutrition . Surgery in the hepatobiliary pancreatic field is known to have a high rate of complications . Uninterrupted perioperative nutrition, i.e., nutrition during the night before, during surgery, and immediately after, offers a strong tool to prevent complications . It is essential that the nutrition also provides food for the colon, e.g., fiber and healthy bacteria (probiotics) to ferment the fiber and boost the immune system.

Berl Munch Tierarztl Wochenschr, 2002 Nov-Dec, 115(11-12), 453 - 7
Effect of Tween 80 and monensin on ruminal fermentation of the diet containing 70% wheat straw treated by white-rot fungus in artificial rumen; Jalc D; The objective of this study was to investigate the effect of Tween 80 and monensin on rumen fermentation of the diet containing 70% wheat straw treated by white-rot fungus Pleurotus tuber-regium (TWS-PT) and 30% barley in artificial rumen (RUSITEC) . The RUSITEC consisted of four fermentation vessels (V1, V2, V3, V4): V1 was without additives (control), V2 received daily 10 mg of monensin, V3 received daily 0.5% Tween 80 (vol.wt-1) and V4 involved the combination of 10 mg of monensin with 0.5% Tween 80 (vol.wt-1) . After an adaptation period (6 days) the fermentation parameters were determined for six consecutive days . Tween 80 did not affect the rumen fermentation of the diet consisting 70% TWS-PT and 30% barley in RUSITEC . Monensin affected the rumen fermentation of the diet by the decreased degradability of DM, neutral detergent fiber (NDF), acid detergent fiber (ADF), hemicellulose, cellulose (p < 0.001), the decrease of methane production (p < 0.001) and the higher proportion of propionate within the volatile fatty acids (p < 0.001) in comparison to control . Tween 80 did not improve the potency of monensin . Only some indices of the increase mol% of propionate (about 3.4%) and the decrease of methane production (about 0.47 mmol.day-1) were found by using Tween 80 plus monensin in comparison to use of monensin alone.

Trends Biotechnol, 2003 Jan, 21(1), 29 - 37
Accelerated design of bioconversion processes using automated microscale processing techniques; Lye GJ et al.; Microscale processing techniques are rapidly emerging as a means to increase the speed of bioprocess design and reduce material requirements . Automation of these techniques can reduce labour intensity and enable a wider range of process variables to be examined . This article examines recent research on various individual microscale unit operations including microbial fermentation, bioconversion and product recovery techniques . It also explores the potential of automated whole process sequences operated in microwell formats . The power of the whole process approach is illustrated by reference to a particular bioconversion, namely the Baeyer-Villiger oxidation of bicyclo{3.2.0}hept-2-en-6-one for the production of optically pure lactones.

Surg Neurol, 2002 Sep-Oct, 58(3-4), 271 - 3
Gas-containing otogenic brain abscess; Paolini S et al.; BACKGROUND: Gas-containing brain abscesses are very rare . Two mechanisms may be responsible for the presence of intracavitary gas: bacterial fermentation or penetration through an abnormal communication between the exterior and the intracranium . The need to search for this potential communication is considered an indication for open surgery . We report the case of a surgically treated gas-containing brain abscess originating from an undiagnosed chronic otitis media . CASE DESCRIPTION: A 54-year-old man developed acute neurologic deterioration, becoming comatose within 24 hours . A contrast-enhanced computed tomography (CT) scan disclosed a gas-containing cystic mass in the right temporal lobe . Urgent surgical decompression revealed the presence of an abscess, which was excised . During the same surgery, we performed a radical mastoidectomy, removing a previously undiagnosed attic cholesteatoma . Neither procedure revealed a discontinuity of the floor of the middle cranial fossa . Cultures grew a mixed flora . Antibiotics were administered for 6 weeks . The patient made a complete neurologic recovery . CONCLUSION: This report demonstrates that otogenic brain abscesses may contain gas due to fermentation of nonclostridial bacteria.

Yeast, 2002 Dec, 19(16), 1399 - 411
Characterization of a gene encoding tRNA nucleotidyltransferase from Candida glabrata; Hanic-Joyce PJ et al.; A gene encoding ATP (CTP):tRNA nucleotidyltransferase (EC2.7.7.25) was isolated from Candida (Torulopsis) glabrata by complementation in Saccharomyces cerevisiae . The predicted amino acid sequence of the protein revealed a large region with high sequence similarity to members of the Class II group of the nucleotidyltransferase superfamily and an N-terminal region characteristic of a mitochondrial targeting sequence . The essential role of the carboxylates within the conserved DXD and RRD motifs was confirmed by mutagenesis . C . glabrata strains bearing truncated CCA1 genes that lacked sequences encoding the putative mitochondrial targeting peptide were unable to grow on non-fermentable carbon sources but were able to grow on a fermentable carbon source . These results suggest that, as in S . cerevisiae, the C . glabrata CCA-adding enzyme is a sorting isozyme that functions in multiple cellular compartments . Mapping of the 5'-ends of primary transcripts of CCA1 revealed multiple transcription start sites located both upstream of and between two in-frame start codons . When the cells were cultured on a non-fermentable carbon source the longer transcripts appeared more abundant, suggesting that the choice of transcription start sites was influenced by carbon source . The shorter transcripts, which lacked sequences encoding the mitochondrial targeting information, were more predominant in cells grown on glucose . These observations suggest that expression of CCA-adding isozymes in C . glabrata may be regulated . The DNA sequence has been assigned GenBank Accession No . AF098803 .

Curr Genet, 2002 Nov, 42(2), 94 - 102 Epub 2002 Nov 21.
The mitochondrial genome can be altered or lost without lethal effect in the petite-negative yeast Debaryomyces (Schwanniomyces) occidentalis; Fernet CS et al.; The nature of mutations affecting several cytochrome-deficient mutants of Debaryomyces (Schwanniomyces) occidentalis has been characterized . The DR12 mutant, which is deficient in cytochrome b, and the B10Mn mutant, which is deficient in cytochromes b and a, a3, are deleted in the mitochondrial CYB and COX1 genes respectively . The B10 strain, which is partially deficient in cytochrome b, has no detectable change in its mitochondrial DNA and possibly carries nuclear lesion(s) . These three mutants, unlike the rho(-) and rho degrees "petite" mutants of Saccharomyces cerevisiae, can still grow on non-fermentable substrates, due to the development of a salicylhydroxamic acid (SHAM)-sensitive alternative pathway linked to phosphorylation at site 1 . A gly(-) mutant lacking mtDNA and respiratory capacity has been isolated . For the first time, it is demonstrated that mtDNA can be altered or even lost without lethal consequence in D . occidentalis, although this yeast was classified as a petite-negative species.

Microbiol Immunol, 2002, 46(10), 667 - 75
Extracellular ATP regulates cell death of lymphocytes and monocytes induced by membrane-bound lipoproteins of Mycoplasma fermentans and Mycoplasma salivarium; Into T et al.; The cytotoxicities of lipoproteins of Mycoplasma fermentans and Mycoplasma salivarium to a lymphocytic cell line, MOLT-4, and a monocytic cell line, HL-60, was upregulated by ATP added extracellularly in a dose-dependent manner . These lipoproteins induced ATP release and plasma membrane permeability increase in these cell lines . In addition, periodate-oxidized ATP, an antagonist for P2X purinergic receptors, suppressed the cytotoxicity of the lipoproteins, suggesting the possibility that P2X receptors for ATP play crucial roles in the cytotoxicity . Activation of caspase-3 induced by the lipoproteins, which was assessed by the cleavage of the synthetic substrate DEVD-pNA and the endogenous substrate poly(ADP-ribose) polymerase, was also upregulated and downregulated by extracellular ATP and periodate-oxidized ATP, respectively . On the basis of these results, this study suggests that mycoplasmal lipoproteins induce the permeability increase in lymphocytes and monocytes, by which ATP is released, and the ATP regulates the cytotoxicities of the lipoproteins to the cells, possibly by interaction with ATP receptors such as P2X purinergic receptors.

Mikrobiyol Bul, 2002 Jan, 36(1), 49 - 55
{Catalase-negative Rothia dentocariosa: evaluation of additional descriptive tests}; Ergin C et al.; Rothia dentocariosa is one of the human oral flora members, and classified in the group of coryneform bacteria which are taxonomically heterogeneous . Although R . dentocariosa is considered as a contaminant in throat cultures generally, it may be the causative infectious agent in the presence of predisposing factors . The catalase activity which is used as a simple and initial identification test, becomes contradictory because of the detection of the catalase negative strains recently . The aim of this study was to compare several biochemical and enzymatic reactions for the identification of catalase positive and negative R . dentocariosa strains . A total of 42 R . dentocariosa strains which were isolated from throat cultures have been studied, and the positivity rate of mannose fermentation in catalase negative R . dentocariosa isolates was found to be significantly higher than those catalase positive (p < 0.05) . In conclusion, mannose fermentation test seems to be a useful tool as an additional identification method of catalase negative R . dentocariosa isolates.

J Agric Food Chem, 2002 Dec 18, 50(26), 7660 - 8
Volatile response of four apple varieties with different coatings during marketing at room temperature; Bai J et al.; Five experimental coatings with different resistance to gas exchange were used with freshly harvested and 20-week commercially stored apples of Delicious, Fuji, Braeburn, and Granny Smith varieties . The coated or noncoated apples were held at 20 degrees C for up to 4 weeks . The gas partial pressures inside the fruits with the various coatings ranged from 1 to 25 kPa CO(2) and from 20 to 1 kPa O(2) . Volatile evaporation rates were measured, as also were the volatiles compositions in the fruit . The coatings with intermediate gas resistance (carnauba-shellac mixture and candelilla) gave intermediate values of CO(2) and O(2) in the internal atmosphere in Delicious, Fuji, and Braeburn apples and the highest concentrations of butyl acetate and 2-methylbutyl acetate in the fruits . The coatings with the highest gas resistance (shellac and shellac-protein) caused high internal CO(2) and low O(2), resulting in anaerobic fermentation in Braeburn and Granny Smith apples and relatively high amounts of low-molecular-weight ethyl esters trapped within the fruit . A small portion of the alcohols were evaporated from fruits compared to esters, this attributed to their high Henry's law coefficients.

Biotechnol Bioeng, 2003 Feb 5, 81(3), 341 - 7
Pulsed feeding during fed-batch fungal fermentation leads to reduced viscosity without detrimentally affecting protein expression; Bhargava S et al.; The goal in this study was to determine if pulsed addition of substrate could be used to alter filamentous fungal morphology during fermentation, to result in reduced broth viscosity . In all experiments, an industrially relevant strain of Aspergillus oryzae was grown in 20-liter fermentors . As a control, cultures were fed limiting substrate (glucose) continuously . Tests were performed by altering the feeding strategy so that the same total amount of glucose was fed in repeated 300-s cycles, with the feed pump on for either 30 or 150 s during each cycle . Variables indicative of cellular metabolic activity (biomass concentration, oxygen uptake rate, base consumed for pH control) showed no significant difference between continuous and pulse-fed fermentations . In addition, there was no significant difference between total extracellular protein expression or the apparent distribution of these proteins . In contrast, fungal mycelia during the second half of pulse-fed fermentations were approximately half the size (average projected area) of fungi during fermentations with continuous addition of glucose . As a result, broth viscosity during the second half of pulse-fed fermentations was approximately half that during the second half of continuous fermentations . If these results prove to be applicable for other fungal strains and processes, then this method will represent a simple and inexpensive means to reduce viscosity during filamentous fungal fermentation .

Arch Microbiol, 2002 Dec, 179(1), 1 - 6 Epub 2002 Nov 07.
Dissimilation of the C2 sulfonates; Cook AM et al.; Organosulfonates are widespread in the environment, both as natural products and as xenobiotics; and they generally share the property of chemical stability . A wide range of phenomena has evolved in microorganisms able to utilize the sulfur or the carbon moiety of these compounds; and recent work has centered on bacteria . This Mini-Review centers on bacterial catabolism of the carbon moiety in the C2-sulfonates and the fate of the sulfonate group . Five of the six compounds examined are subject to catabolism, but information on the molecular nature of transport and regulation is based solely on sequencing data . Two mechanisms of desulfonation have been established . First, there is the specific monooxygenation of ethanesulfonate or ethane-1,2-disulfonate . Second, the oxidative, reductive and fermentative modes of catabolism tend to yield the intermediate sulfoacetaldehyde, which is now known to be desulfonated to acetyl phosphate by a thiamin-diphosphate-dependent acetyltransferase . This enzyme is widespread and at least three subgroups can be recognized, some of them in genomic sequencing projects . These data emphasize the importance of acetyl phosphate in bacterial metabolism . A third mechanism of desulfonation is suggested: the hydrolysis of sulfoacetate.

J Environ Qual, 2002 Nov-Dec, 31(6), 1795 - 801
Application technique and slurry co-fermentation effects on ammonia, nitrous oxide, and methane emissions after spreading: II . Greenhouse gas emissions; Wulf S et al.; The aim of this study was to investigate the effect of different application techniques on greenhouse gas emission from co-fermented slurry . Ammonia (NH3), nitrous oxide (N2O), and methane (CH4) emissions were measured in two field experiments with four different application techniques on arable and grassland sites . To gather information about fermentation effects, unfermented slurry was also tested, but with trail hose application only . Co-fermented slurry was applied in April at a rate of 30 m3 ha(-1) . Measurements were made every 4 h on the first day after application and were continued for 6 wk with gradually decreasing sampling frequency . Methane emissions were <150 g C ha(-1) from co-fermentation products and seemed to result from dissolved CH4 . Only in the grassland experiment were emissions from unfermented slurry significantly higher, with wetter weather conditions probably promoting CH4 production . Nitrous oxide emission was significantly increased by injection on arable and grassland sites two- and threefold, respectively . Ammonia emissions were smallest after injection or trail shoe application and are discussed in the preceding paper . We evaluated the climatic relevance of the measured gas emissions from the different application techniques based on the comparison of CO2 equivalents . It was evident that NH3 emission reduction, which can be achieved by injection, is at least compensated by increased N2O emissions . Our results indicate that on arable land, trail hose application with immediate shallow incorporation, and on grassland, trail shoe application, bear the smallest risks of high greenhouse gas emissions when fertilizing with co-fermented slurry.

J Environ Qual, 2002 Nov-Dec, 31(6), 1789 - 94
Application technique and slurry co-fermentation effects on ammonia, nitrous oxide, and methane emissions after spreading: I . Ammonia volatilization; Wulf S et al.; Ammonia emissions after spreading animal manure contribute a major share to N losses from agriculture . There is an increasing interest in anaerobic co-digestion of liquid manure with organic additives . This fermentation results in a change of physical and chemical parameters of the slurry . Among these are an increased pH and ammonium content, implying a higher risk of NH3 losses from fermentation products . To compare different application techniques and the effect of fermentation on NH3 volatilization, we used the standard comparison method and tested it for reliability . This method seems to be perfectly suited for experiments with a large number of treatments and replicates if prerequisites concerning the experimental layout are considered . We tested four different application techniques on arable and grassland sites . The more the substrate was incorporated into the soil or applied near the soil surface on the grassland site, the less NH3 was lost . Injection of the substrate reduced losses to less than 10% of applied NH4+ on both sites, whereas losses after splash plate application amounted to more than 30% . Trail shoe application on grassland performed as well as injection . Harrowing on arable land also reduced emissions efficiently, if harrowing occurred within the first 2 h after application . Emissions from trail hose-applied co-fermentation product were not greater than from unfermented slurry . Better infiltration of the less viscous substrate seemed to have compensated for the increased loss potential.

J Nutr, 2002 Dec, 132(12), 3599 - 602
Dietary chicory inulin increases whole-body bone mineral density in growing male rats; Roberfroid MB et al.; Chicory inulin is a natural linear fructan that is not digested in the upper part of the gastrointestinal tract but is fermented in the cecocolon . It enhances calcium absorption in rats and improves femur and tibia mineral contents in gastrectomized or ovariectomized rats . We studied the effect of inulin (0, 5 and 10 g/100 g diet) on whole-body bone mineral content (WBBMC), whole-body bone area (WBBA) and whole-body bone mineral density (WBBMD) in live, growing male rats fed diets containing 0.2, 0.5 or 1 g Ca/100 g . Three experiments, each corresponding to one of the different dietary Ca concentrations, were performed using male Wistar rats (n = 108; 4 wk old) . WBBMC was measured by dual-energy X-ray absorptiometry every 4 wk up to wk 22 . Inulin increased WBBMC (P < 0.05) and WBBMD (P < 0.001) significantly but not WBBA at all ages and all dietary calcium concentrations . This is the first report to demonstrate that chicory inulin not only increases calcium absorption but also increases mineral parameters in whole-body bones.

Biotechnol Prog, 2002 Nov-Dec, 18(6), 1431 - 8
Comparison of techniques for monitoring antibody fragment production in E . coli fermentation cultures; Bowering LC et al.; The use of an optical biosensor for monitoring antibody fragment accumulation following induction in a batch fermentation of recombinant E . coli is compared to the more traditional method of ELISA quantification . Using the biosensor, concentration data can be obtained within minutes of sample addition to the device, compared to an average assay time of 3-4 h for the ELISA . We describe two biosensor assays developed as an alternative to ELISA and compare them with ELISA in the ability to provide quantitative product accumulation profiles during fermentation . Discrepancies in titers recorded by the assays are explained by a combination of differences in product variants detected by each assay and interference from sample contaminants . Method of sample preparation is also shown to be important if accurate concentration data is required . Both biosensor assays are shown to be capable of providing product accumulation profiles comparable to those obtained by ELISA . The use of a rapid extraction technique would allow such data to be obtained during process operation, enabling improved fermentation control and more rapid process development.

Biotechnol Prog, 2002 Nov-Dec, 18(6), 1366 - 76
Identification of critical batch operating parameters in fed-batch recombinant E . coli fermentations using decision tree analysis; Buck KK et al.; To develop a useful fermentation process model, it is first necessary to identify which batch operating parameters are critical in determining the process outcome . To identify critical processing inputs in large databases, we have explored the use of Decision Tree Analysis with the decision metrics of Gain (i.e., Shannon Entropy changes), Gain Ratio, and a multiple hypergeometric distribution . The usefulness of this approach lies in its ability to treat "categorical" variables, which are typical of archived fermentation databases, as well as "continuous" variables . In this work, we demonstrate the use of Decision Tree Analysis for the problem of optimizing recombinant green fluorescent protein production in E . coli . A database of 85 fermentations was generated to examine the effect of 15 process input parameters on final biomass yield, maximum recombinant protein concentration, and productivity . The use of Decision Tree Analysis led to a considerable reduction in the fermentation database through the identification of the significant as well as insignificant inputs . However, different decision metrics selected different inputs and different numbers of inputs to classify the data for each output.

Biotechnol Prog, 2002 Nov-Dec, 18(6), 1356 - 65
Genetic programming assisted stochastic optimization strategies for optimization of glucose to gluconic acid fermentation; Cheema JJ et al.; This article presents two hybrid strategies for the modeling and optimization of the glucose to gluconic acid batch bioprocess . In the hybrid approaches, first a novel artificial intelligence formalism, namely, genetic programming (GP), is used to develop a process model solely from the historic process input-output data . In the next step, the input space of the GP-based model, representing process operating conditions, is optimized using two stochastic optimization (SO) formalisms, viz., genetic algorithms (GAs) and simultaneous perturbation stochastic approximation (SPSA) . These SO formalisms possess certain unique advantages over the commonly used gradient-based optimization techniques . The principal advantage of the GP-GA and GP-SPSA hybrid techniques is that process modeling and optimization can be performed exclusively from the process input-output data without invoking the detailed knowledge of the process phenomenology . The GP-GA and GP-SPSA techniques have been employed for modeling and optimization of the glucose to gluconic acid bioprocess, and the optimized process operating conditions obtained thereby have been compared with those obtained using two other hybrid modeling-optimization paradigms integrating artificial neural networks (ANNs) and GA/SPSA formalisms . Finally, the overall optimized operating conditions given by the GP-GA method, when verified experimentally resulted in a significant improvement in the gluconic acid yield . The hybrid strategies presented here are generic in nature and can be employed for modeling and optimization of a wide variety of batch and continuous bioprocesses.

Biotechnol Prog, 2002 Nov-Dec, 18(6), 1141 - 8
Modulation of phosphoenolpyruvate synthase expression increases shikimate pathway product yields in E . coli; Yi J et al.; Product yields in microbial synthesis are ultimately limited by the mechanism utilized for glucose transport . Altered expression of phosphoenolpyruvate synthase was examined as a method for circumventing these limits . Escherichia coli KL3/pJY1.216A was cultured under fed-batch fermentor conditions where glucose was the only source of carbon for the formation of microbial biomass and the synthesis of product 3-dehydroshikimic acid . Shikimate pathway byproducts 3-deoxy-D-arabino-heptulosonic acid, 3-dehydroquinic acid, and gallic acid were also generated . An optimal expression level of phosphoenolpyruvate synthase was identified, which did not correspond to the highest expression levels of this enzyme, where the total yield of 3-dehydroshikimic acid and shikimate pathway byproducts synthesized from glucose was 51% (mol/mol) . For comparison, the theoretical maximum yield is 43% (mol/mol) for synthesis of 3-dehydroshikimic acid and shikimate pathway byproducts from glucose in lieu of amplified expression of phosphoenolpyruvate synthase.

Appl Microbiol Biotechnol, 2002 Dec, 60(4), 455 - 60 Epub 2002 Oct 15.
Use of a histone H4 promoter to drive the expression of homologous and heterologous proteins by Penicillium funiculosum; Belshaw NJ et al.; Two genes encoding histone H4 (H4.1 and H4.2) from Penicillium funiculosum have been cloned and characterised . Structurally, the histone H4.1 gene is divergently linked to the histone H3 gene and the two genes are separated by approximately 800 bp . The transcription of the histone H4.1 and H4.2 genes in P . funiculosum appears to be distinctively regulated . Histone H4.1 mRNA showed a high steady-state level during the early stages of batch culture that decreased as growth reached the stationary phase . In contrast, the expression of the histone H4.2 gene was lower than that of H4.1 throughout batch growth and increased gradually with time . In order to expand the industrial application of P . funiculosum as a host for the production of heterologous proteins, the promoter of the histone H4.1 gene was successfully used to drive the expression of an intracellular bacterial enzyme, beta-glucuronidase, and a secreted homologous enzyme, xylanase C . The constitutive secretion of xylanase C was achieved in the absence of other xylanases by batch fermentation in the presence of glucose.

Environ Technol, 2002 Oct, 23(10), 1147 - 56
Modelling of an activated primary settling tank including the fermentation process and VFA elutriation; Ribes J et al.; A complete model of a primary settler including both sedimentation and biological processes is presented . It is a one-dimensional model based on the solids flux concept and the conservation of mass that uses the Takacs model for the settling velocity, which is corrected by a compression function in the lower layers . The biological model is based on the ASM2 and enlarged with the fermentation model proposed by this research group . The settler was split in ten layers and the flux terms in the mass balance for each layer is obtained by means of the settling model . A pilot plant has been operated to study the primary sludge fermentation and volatile fatty acids (VFA) elutriation in a primary settler tank . The model has been tested with pilot plant experimental data with very good results . It has been able to simulate the VFA production in the settler and their elutriation with the influent wastewater for all the studied experiments . The developed model is easily applicable to secondary settlers and thickeners, also taking into account biological activity inside them.

Nat Prod Lett, 2002 Dec, 16(6), 377 - 82
Microbial transformation of (+)-androsta-1 ,4-diene-3,17-dione by Cephalosporium aphidicola; Choudhary MI et al.; Fermentation of (+)-androsta-1,4-diene-3,17-dione ({structure: see text}) with Cephalosporium aphidicola for 8 days yielded oxidative and reductive metabolites, androst-4-ene-3,17-dione ({structure: see text}), 17beta-hydroxyandrosta-1,4-diene-3-one ({structure: see text}), 11alpha-hydroxyandrosta-1,4-diene-3,17-dione ({structure: see text}), 11alpha-hydroxyandrost-4-ene-3,17-dione ({structure: see text}), 11alpha,17beta-dihydroxyandrost-4-ene-3-one ({structure: see text}) and 11alpha,17beta-dihydroxyandrosta-1,4-diene-3-one ({structure: see text}) . The fermentation of {structure: see text} with Fusarium lini also yielded metabolites {structure: see text} . The structures of these metabolites were elucidated on the basis of spectroscopic techniques.

J Anim Sci, 2002 Nov, 80(11), 3016 - 20
Effect of exogenous fibrolytic enzyme on ruminal fermentation and digestibility of alfalfa and rye-grass hay fed to lambs; Pinos-Rodriguez JM et al.; This experiment was carried out to study the effect of a directly fed exogenous fibrolytic enzyme on intake and digestion of DM, OM, protein, NDF, ADF, and hemicellulose of alfalfa and ryegrass hay by sheep . Four diets were randomly assigned to four ruminally cannulated lambs using a 4 x 4 Latin square design, repeated in time, with a factorial arrangement (2 x 2) of diets: 1) alfalfa hay; 2) alfalfa hay + exogenous fibrolytic enzymes (enzyme); 3) ryegrass hay; and 4) ryegrass hay + enzyme . Lambs consumed more DM and OM from alfalfa than from ryegrass hay (P < 0.001) . The ADF intake was not different between the hays, but NDF intake was lower for alfalfa (P < 0.001) . For both hays, the enzyme increased intake of DM (P < 0.01), as well as OM and CP (P < 0.05); however, NDF and ADF intake were not changed . Alfalfa hay had higher apparent digestibility of DM, OM, and CP (P < 0.001), but lower digestibility for NDF, ADF, and hemicellulose . The enzyme increased apparent digestibility of CP, hemicellulose (P < 0.05), and NDF (P < 0.10) for alfalfa . Also, for both hays, the enzyme improved N balance because lambs retained more N (P < 0.05) . The enzyme increased (P < 0.05) total VFA concentration (3 and 6 h) for both hays . Results from this trial indicate that directly fed exogenous fibrolytic enzymes may change ruminal fermentation, intake, and digestibility of forages with different nutritive value.

J Anim Sci, 2002 Nov, 80(11), 3005 - 15
Effects of virginiamycin and monensin plus tylosin on ruminal protein metabolism in steers fed corn-based finishing diets with or without wet corn gluten feed; Ives SE et al.; Six ruminally cannulated steers (345 +/- 20 kg initial BW) were used in a 6 x 6 Latin square to evaluate effects of diet and antibiotics on ruminal protein metabolism . Two diets and three antibiotic treatments were arranged factorially . One diet contained (DM basis) 72% dry-rolled corn, 12% soybean meal, 10% alfalfa hay, and 4% molasses (SBM), and the other contained 63% dry-rolled corn, 30% wet corn gluten feed, and 5% alfalfa hay (WCGF) . Antibiotic treatments included control, virginiamycin (175 mg/d; VM), and monensin/tylosin (250 and 100 mg/d, respectively; MT) . Steers were fed at 12-h intervals at a rate of 2.4% of empty BW daily . Each period included 18 d of adaptation and 3 d of ruminal fluid collections . Samples were collected at 0, 2, 4, 6, 8, and 10 h after the morning feeding on d 19 and 20 . On d 21, rumens were dosed 2 h after the morning feeding with 350 g of solubilized casein to evaluate in vivo ruminal protease and deaminase activities . Ruminal fluid samples were collected 1, 2, 3, 4, and 6 h after the casein dose . On d 19 and 20, antibiotics had no effect on ruminal pH or concentrations of VFA, lactate, ammonia, ciliated protozoa, alpha-amino nitrogen (AAN), or peptide N, but VM reduced (P < 0.01) the concentration of isovalerate compared to MT and control . After casein dosing (d 21), peptide N concentration was unaffected by antibiotics, but AAN were higher (P < 0.01) for VM than MT and control . Relative to MT and control, VM reduced ruminal isovalerate (P = 0.05) and increased ruminal propionate (P < 0.01) on d 21 . Ruminal pH was lower (P < 0.01) in steers fed SBM than in steers fed WCGF, but lactate concentrations were unaffected by diet . Steers fed SBM had higher (P < 0.05) ruminal concentrations of total VFA and propionate . Ammonia concentrations were lower before feeding and higher after feeding for steers fed WCGF (P < 0.01) . Steers fed WCGF had higher counts of total ciliated protozoa than steers fed SBM (P < 0.05) due to greater Entodinium sp . (P < 0.05) . Steers fed WCGF had higher (P < 0.01) ruminal AAN and peptide N concentrations than those fed SBM on d 19 and 20 . After casein dosing, ruminal peptide N concentrations were similar, but AAN were lower (P < 0.01) for WCGF than SBM . Overall, VM appeared to depress ruminal deaminase activity, and MT had minimal effects on ruminal fermentation products . The protein in WCGF appeared to be more readily degradable than that in SBM.

J Anim Sci, 2002 Nov, 80(11), 2978 - 88
Influence of rumen protein degradability and supplementation frequency on steers consuming low-quality forage: II . Ruminal fermentation characteristics; Bohnert DW et al.; Seven ruminally and duodenally cannulated steers (264 +/- 8 kg BW) consuming low-quality forage (5% CP; 61% NDF; 31% ADF) were used to determine the influence of CP degradability and supplementation frequency (SF) on ruminal fermentation characteristics . Treatments included an unsupplemented control and degradable intake protein (DIP) or undegradable intake protein (UIP) provided daily, every 3 d, or every 6 d . The DIP treatments (18% UIP) were calculated to provide 100% of the DIP requirement, while the UIP treatments (60% UIP) were provided on an isonitrogenous basis compared with DIP . Ruminal NH3-N was increased on the day all supplements were provided with supplemental CP (P = 0.04) and for DIP compared with UIP (P < 0.01) . Also, because ruminal NH3-N increased at a greater rate with DIP compared with UIP as SF decreased, a linear effect of SF x CP degradability interaction (P = 0.02) was observed . In addition, NH3-N was greater on the day only daily supplements were provided for supplemented treatments (P = 0.04), and decreased linearly (P < 0.01) as SF decreased . Concentration of total VFA increased linearly (P = 0.02) as SF decreased on the day all supplements were provided, whereas on the day only daily supplements were provided, total VFA were greater for UIP compared with DIP (P = 0.01), and decreased linearly (P < 0.01) as SF decreased . An interaction concerning the linear effect of SF and CP degradability (P = 0.02) was observed for ruminal liquid volume on the day all supplements were provided . This was the result of an increase in liquid volume with DIP as SF decreased compared with a minimal effect with UIP . In contrast, there was no influence of supplementation on liquid volume the day only daily supplements were provided . Ruminal liquid dilution rate was greater (P = 0.02) with CP supplementation on the day all supplements were provided . We did observe a quadratic effect of SF x CP degradability interaction (P = 0.01) for dilution rate because of a quadratic response with DIP (greatest value with the every-third-day treatment) compared with a decrease as SF decreased for UIP . On the day only daily supplements were provided, ruminal liquid dilution rate decreased linearly (P = 0.02) as SF decreased . These results suggest that DIP and UIP elicit different effects on ruminal fermentation when supplemented infrequently to ruminants consuming low-quality forage while not adversely affecting nutrient intake and digestibility.

Lett Appl Microbiol, 2002, 35(6), 473 - 80
Investigating the prebiotic and gas-generating effects of selected carbohydrates on the human colonic microflora; Probert HM et al.; AIMS: To compare the fermentation of dietary carbohydrates with reference to their prebiotic and gas-generating capacity . METHODS AND RESULTS: Static anaerobic batch culture fermentations were carried out measuring gas generation and the prebiotic effect of five selected substrates (including various fructo-oligosaccharides, levan and maltodextrin) . The largest gas producer was levan, whilst those showing no significant difference to Actilight included oligofructose and maltodextrin . Gas composition data showed that hydrogen and carbon dioxide were the two most quantitatively important gases . The substrate that appeared to have the best prebiotic effect in vitro was branched chain fructo-oligosaccharide (FOS), followed by oligofructose, Actilight and maltodextrin which each exerted a similar effect . The substrate with the least bifidogenic effect was levan . CONCLUSIONS: The composition and total gas generation data showed that there was much variation between and within donor inocula . Generally, the lower gas producers had a more selective fermentation whilst larger gas producers were less specific . SIGNIFICANCE AND IMPACT OF THE STUDY: The study of these three parameters enabled a more complete picture of carbohydrate breakdown to be drawn and hence highlighted the need for potential prebiotics to be more extensively evaluated in order to reduce negative side-effects such as gas distension.

Caries Res, 2002 Nov-Dec, 36(6), 417 - 22
Evaluation of the cariogenic potential of cassava flours from the Amazonian region; Rebelo Vieira JM et al.; Cassava flour is the main dietary carbohydrate source in the Amazonian region . The cariogenic potentials of the two main kinds of cassava flour - 'seca' and 'd'agua' - were evaluated in vivo and in vitro . A sweet made from a regional fruit (cupuacu) and a species of local fish (tambaqui) were used as positive and negative controls, respectively . For in vivo evaluation of dental plaque acidogenicity, the study had a crossover design, in four legs, with 19 volunteers in four treatment groups . Dental plaque pH was determined by the microtouch method before and for 60 min after food consumption . For in vitro evaluation, stimulated saliva of each volunteer was incubated with the food and pH variation was determined over 4 h . Carbohydrates were chemically determined in the flours and the majority was found to be water-insoluble . In vivo, plaque pH decreased significantly after the consumption of the sweet (p < 0.05), increased after the fish (p < 0.05), but did not change after intake of the flours (p > 0.05) . In vitro, the flours were slowly fermented by bacteria present in saliva . The in vivo and in vitro findings suggest that, in the form that the main cassava flours from the Amazonian region of Brazil are customarily eaten, they may be considered to have no or very low cariogenicity .

J Microbiol Methods, 2003 Feb, 52(2), 239 - 44
Selective enrichment and purification of cultures of Methanosaeta spp; Janssen PH; A simple method for the isolation of axenic cultures of members of the obligately acetotrophic methanogenic genus Methanosaeta is described . To overcome the competitive advantage obtained by faster growing acetate-utilizing Methanosarcina spp . in batch enrichment cultures, acetone and isopropanol are used as the growth substrates for the enrichment step . Acetone- and isopropanol-utilizing bacteria slowly ferment these substrates to acetate, which allows Methanosaeta spp . to maintain the acetate concentration at levels below the threshold required for growth of Methanosarcina spp . These enrichments eventually develop dense populations of Methanosaeta spp., which can then be separated from contaminating microorganisms to yield axenic cultures .

J Chromatogr A, 2002 Nov 29, 978(1-2), 153 - 64
Immobilized metal-ion affinity chromatography of recombinant Fab protein OPG C11 in the presence of EDTA-Mg(II); Xiang H et al.; Undesired adsorption of host cell proteins poses a big challenge for immobilized metal-ion affinity chromatography (IMAC) purification . In this study, by using His6-tagged protein Fab OPG C11 from Escherichia coli fermentation as a model, we found that the presence of low concentrations of EDTA-Mg2+ in feed streams weakens the adsorption but makes it more specific towards polyhistidine tag . By combining EDTA-Mg2+ treatment and periplasmic extraction, we developed a one-step purification procedure for His6-tagged recombinant Fab OPG C11 using Ni-IDA (iminodiacetic acid) chromatography . This procedure eliminated the buffer exchange step after periplasmic extraction, which is usually required before IMAC in order to remove EDTA . In addition to savings on time and cost, this procedure eliminates undesired adsorption of most host cell proteins thus significantly improves the purity of polyhistidine-tagged recombinant proteins . The strategy of EDTA-Mg2+ treatment may have general application potentials.

J Antibiot (Tokyo), 2002 Sep, 55(9), 789 - 93
Nigerloxin, a novel inhibitor of aldose reductase and lipoxygenase with Free radical scavenging activity from Aspergillus niger CFR-W-105; Rao KC et al.; An enzyme inhibitor, nigerloxin, with inhibition against soy bean lipoxygenase-I (LOX-1), rat lens aldose reductase (RLAR) as well as free radical scavenging activity was isolated from the fermented wheat bran using Aspergillus niger CFR-W-105 . Its chemical structure was identified as 2-amido-3-hydroxy-6-methoxy-5-methyl-4-(prop-1'-enyl) benzoic acid by NMR and GCEIMS data . The IC50 values against LOX-1 and RLAR were found to be 79 microM and 69 microM and ED50 against 1,1-diphenyl-2-picrylhydrazyl (DPPH) was 66 microM.

Vet Hum Toxicol, 2002 Dec, 44(6), 366 - 9
Cyanogenic potential of cassava peels and their detoxification for utilization as livestock feed; Tweyongyere R et al.; This study determined the cyanogenic potential of the cassava peels and assess the effectiveness of sun drying, heap fermentation and wet fermentation (soaking) in reducing the cyanide potential of the peels . Fresh cassava peels from major fresh food markets in Kampala and cassava grown in various parts of Uganda from Namolonge Agricultural and Animal Research Institute were used . The fresh peels from the market were subjected to the different detoxification methods foe 5 d; the cyanide potential was determined by enzymatic assay . The mean potential of the cassava peels from the food markets Kampala was 856 mg cyanide equivalen/kg of dry matter . The potential of the peels of the 14 cultivars fell between 253 and 1081 mg cyanide eQuivalent/kg of dry matter . High cyanogenic potential cultivars dominate on the market and pose danger of poisoning to livestock fed on fresh cassava peels . Treatment of the peels by sun-drying, heap fermentation on soaking reduced the cyanide potential to below 100 mg cyanide equivalent/kg of dry matter at 48, 72 and 96 h respectively . Sun-dying caused an early sharp fall in the cyanide potential, but heap fermentation or soaking gave the lowest residual cyanide after 120 h . Cassava peels could be safely used as livestock feed if they are treated to reduce the cyanogenic potential.

J Chromatogr B Analyt Technol Biomed Life Sci, 2002 Dec 25, 782(1-2), 227 - 43
Global analysis of a "simple" proteome: Methanococcus jannaschii; Giometti CS et al.; The completed genome of Methanococcus jannaschii, including the main chromosome and two extra-chromosomal elements, predicts a proteome comprised of 1783 proteins . How many of those proteins are expressed at any given time and the relative abundance of the expressed proteins, however, cannot be predicted solely from the genome sequence . Two-dimensional gel electrophoresis coupled with peptide mass spectrometry is being used to identify the proteins expressed by M . jannaschii cells grown under different conditions as part of an effort to correlate protein expression with regulatory mechanisms . Here we describe the identification of 170 of the most abundant proteins found in total lysates of M . jannaschii grown under optimal fermentation conditions . To optimize the number of proteins detected, two different protein specific stains (Coomassie Blue R250 or silver nitrate) and two different first dimension separation methods (isoelectric focusing or nonequilibrium pH gradient electrophoresis) were used . Thirty-two percent of the proteins identified are annotated as hypothetical (21% conserved hypothetical and 11% hypothetical), 21% are enzymes involved in energy metabolism, 12% are proteins required for protein synthesis, and the remainder include proteins necessary for intermediary metabolism, cell division, and cell structure . Evidence of post-translational modification of numerous M . jannaschii proteins has been found, as well as indications of incomplete dissociation of protein-protein complexes . These results demonstrate the complexity of proteome analysis even when dealing with a relatively simple genome.

Curr Drug Targets Infect Disord, 2001 Nov, 1(3), 249 - 62
New approaches for antigen discovery, production and delivery: vaccines for veterinary and human use; Potter AA et al.; Vaccination of individuals has been practiced for many years and has been one of the most effective methods of controlling infectious diseases . Unfortunately, even with this success, society continues to suffer multi-billion dollar economic losses annually due to infectious diseases . These losses occur in all animal species as well as in humans . In order to further reduce these losses, academicians and companies are employing the multidisciplinary approach to develop better and safer vaccines . These include capitalizing on advances in molecular biology, chemistry, pharmacy, immunology, genomics, proteomics, and fermentation . Thus, we are moving from a more empirical approach to vaccine production to a more focused, and, hopefully, more logical approach to identification and production of protective antigens . Furthermore, formulation and delivery of these antigens in playing a major role in revolutionizing how we deliver vaccines to induce the most appropriate immune response and ensure protection . The current review summarizes some of these advances and speculates as to how future vaccines will be produced and delivered for the benefit of society.

J Agric Food Chem, 2002 Dec 4, 50(25), 7249 - 57
Low-temperature brewing using yeast immobilized on dried figs; Bekatorou A et al.; Dried figs, following exhaustive extraction of their residual sugars with water, were used for immobilization of Saccharomyces cerevisiae AXAZ-1 . The immobilized biocatalyst was used in repeated batch fermentations of glucose at 30 degrees C, where significant reduction of the fermentation time was observed, falling from 65 h in the first batch to 7 h after the sixth batch . Repeated fermentations of wort at room and low temperatures resulted in fermentation times that fell from 26 to 20 h and from 27 to 24 days at 18 and 3 degrees C, respectively . Ethanol and beer productivities were high, showing suitability of the biocatalyst for low-temperature brewing . Diacetyl concentrations were low (0.3-0.5 mg/L), and polyphenols were lower than in commercial products and decreased as the fermentation temperature was decreased (126-50 mg/L) . Ethyl acetate concentrations increased from 53 to 88 mg/L as the temperature was decreased, while the concentration of amyl alcohols at 3 degrees C (58 mg/L) was lower than half of that at 18 degrees C (125 mg/L) . The beers produced at the end of the main fermentation had a fine clarity and a special fruity figlike aroma and taste, distinct from commercial products and more intense than beers produced by cells immobilized on other food-grade supports (gluten pellets or delignified cellulosic materials) . GC-MS analysis did not show significant differences in the qualitative composition of the aroma compounds of the beers produced by immobilized and free cells.

J Agric Food Chem, 2002 Dec 4, 50(25), 7231 - 4
Whey liquid waste of the dairy industry as raw material for potable alcohol production by kefir granules; Athanasiadis I et al.; Kefir granular biomass was used in the fermentation of sweet whey and proved to be more effective compared to single-cell biomass of kefir yeast . The operational stability of the biocatalyst was assessed by carrying out 20 repeated batch fermentations . Levels of ethanol productivity reached 2.57 g L(-1) h(-1)), whereas the yield was 0.45 g/g . The fermentation time was only 8 h . Mixtures of sweet whey with molasses were fermented at initial densities ranging from 4.2 to 10.2 degrees Be and resulted in ethanol yield factors between 0.36 and 0.48 g of ethanol/g of utilized sugar . Lower degrees Be values led to an increase of percentages of ethyl acetate on total volatiles determined and a reduction of amyl alcohols . The addition of 1% black raisin extract to whey appears to promote whey fermentation, whereas the same was not observed in the case of white sultana extract addition . It was finally established that it is preferable to ferment mixtures of whey-molasses by adding molasses in whey after the completion of whey fermentation.

Naturwissenschaften, 2002 Nov, 89(11), 483 - 96 Epub 2002 Oct 25.
Solid and liquid residues as raw materials for biotechnology; Gallert C et al.; In the past few decades huge amounts of solid and paste-like wastes of domestic and industrial origin have been deposited on sanitary landfills worldwide . Only a small proportion was incinerated, where incineration plants were available . Since primary resources, such as ores for metal production or crude oil for the production of gasoline, diesel, solvents and plastics, or coal and natural gas as sources for energy or chemicals are not available in unlimited quantities, and because the deposition of residues, wastes and worn-out commodities on sanitary landfills causes pollution of the atmosphere, the soil and the groundwater due to hazardous gaseous emissions and toxic leachates, wastes from households and from industry must be avoided or minimized at an early stage . Whenever waste material can be recycled it must be re-introduced into production processes and the non-recyclable fractions should be used as a fuel for energy recovery . After incineration, the highly toxic dust fractions of ashes and slags resulting from burning the wastes should be deposited on sanitary landfills, while the granulated mineral slag fractions could be used as a substitute for the sand in cement as a construction material.Here we review various processes for the treatment of organic fractions of differently composed wastes to upgrade them to more valuable, re-usable products or at least to recover their energy content . Upgrading processes of organic wastes include composting, biogas fermentation, production of organic acids and solvents, and biopolymer or biosurfactants production . We also include biological purification procedures for the most important components of wastes, such as chitin from the shells of Crustaceae . Typical examples from pilot-scale or full-scale studies are discussed for each process.

Appl Environ Microbiol, 2002 Dec, 68(12), 6263 - 72
Genetic changes to optimize carbon partitioning between ethanol and biosynthesis in ethanologenic Escherichia coli; Underwood SA et al.; The production of ethanol from xylose by ethanologenic Escherichia coli strain KO11 was improved by adding various medium supplements (acetate, pyruvate, and acetaldehyde) that prolonged the growth phase by increasing cell yield and volumetric productivity (approximately twofold) . Although added pyruvate and acetaldehyde were rapidly metabolized, the benefit of these additives continued throughout fermentation . Both additives increased the levels of extracellular acetate through different mechanisms . Since acetate can be reversibly converted to acetyl coenzyme A (acetyl-CoA) by acetate kinase and phosphotransacetylase, the increase in cell yield caused by each of the three supplements is proposed to result from an increase in the pool of acetyl-CoA . A similar benefit was obtained by inactivation of acetate kinase (ackA), reducing the production of acetate (and ATP) and sparing acetyl-CoA for biosynthetic needs . Inactivation of native E . coli alcohol-aldehyde dehydrogenase (adhE), which uses acetyl-CoA as an electron acceptor, had no beneficial effect on growth, which was consistent with a minor role for this enzyme during ethanol production . Growth of KO11 on xylose appears to be limited by the partitioning of carbon skeletons into biosynthesis rather than the level of ATP . Changes in acetyl-CoA production and consumption provide a useful approach to modulate carbon partitioning . Together, these results demonstrate that xylose fermentation to ethanol can be improved in KO11 by redirecting small amounts of pyruvate away from fermentation products and into biosynthesis . Though negligible with respect to ethanol yield, these small changes in carbon partitioning reduced the time required to complete the fermentation of 9.1% xylose in 1% corn steep liquor medium from over 96 h to less than 72 h.

Appl Environ Microbiol, 2002 Dec, 68(12), 6087 - 93
Flow cytometric assessment of membrane integrity of ethanol-stressed Oenococcus oeni cells; Graca da Silveira M et al.; The practical application of commercial malolactic starter cultures of Oenococcus oeni surviving direct inoculation in wine requires insight into the mechanisms involved in ethanol toxicity and tolerance in this organism . Exposure to ethanol resulted in an increase in the permeability of the cytoplasmic membrane, enhancing passive proton influx and concomitant loss of intracellular material (absorbing at 260 nm) . Cells grown in the presence of 8% (vol/vol) ethanol revealed adaptation to ethanol stress, since these cells showed higher retention of compounds absorbing at 260 nm . Moreover, for concentrations higher than 10% (vol/vol), lower rates of passive proton influx were observed in these ethanol-adapted cells, especially at pH 3.5 . The effect of ethanol on O . oeni cells was studied as the ability to efficiently retain carboxyfluorescein (cF) as an indicator of membrane integrity and enzyme activity and the uptake of propidium iodide (PI) to assess membrane damage . Flow cytometric analysis of both ethanol-adapted and nonadapted cells with a mixture of the two fluorescent dyes, cF and PI, revealed three main subpopulations of cells: cF-stained intact cells; cF- and PI-stained permeable cells, and PI-stained damaged cells . The subpopulation of O . oeni cells that maintained their membrane integrity, i.e., cells stained only with cF, was three times larger in the population grown in the presence of ethanol, reflecting the protective effect of ethanol adaptation . This information is of major importance in studies of microbial fermentations in order to assign bulk activities measured by classical methods to the very active cells that are effectively responsible for the observations.

Med Hypotheses, 2003 Jan, 60(1), 65 - 8
Chronic fatigue syndrome: a risk factor for osteopenia?
Nijs J, De Meirleir K, Englebienne P, McGregor N.
No data documenting a possible depletion of bone mineral density in patients with chronic fatigue syndrome (CFS) are currently available . However, recent pathophysiological observations in CFS patients may have deleterious consequences on bone density . Firstly, the deregulation of the 2,5A synthetase RNase L antiviral pathway and its associated channelopathy, implicates increased demands for calcium and consequent increased calcium-re-absorption from the skeletal system . Secondly, Mycoplasma fermentans which has been frequently associated with CFS, produces a lipopeptide, named 2-kDa macrophage-activating lipopeptide (MALP-2), which stimulates macrophages . MALP-2 has been shown to enhance bone resorption in a dose-dependent manner, at least in part by stimulating the formation of prostaglandins . Thirdly, decreased levels of insulin-like growth factor I (IGF-I) have been reported in CFS-patients . IGF-I is critical to the proliferation of osteoblasts . Consequently, depleted levels of IGF-I may shift the balance between osteoclastic and osteoblastic activity towards bone resorption.

J Chromatogr B Analyt Technol Biomed Life Sci, 2002 Dec 5, 781(1-2), 39 - 56
Metabolic acidosis: separation methods and biological relevance of organic acids and lactic acid enantiomers; Ewaschuk JB et al.; Metabolic acidosis can result from accumulation of organic acids in the blood due to anaerobic metabolism or intestinal bacterial fermentation of undigested substrate under certain conditions . These conditions include short-bowel syndrome, grain overfeeding of ruminants and, as recently reported, severe gastroenteritis . Measuring fermentation products such as short-chain fatty acids (SCFAs) and lactic acid in various biological samples is integral to the diagnosis of bacterial overgrowth . Stereospecific measurement of D- and L-lactic acid is necessary for confirmation of the origin and nature of metabolic acidosis . In this paper, methods for the separation of SCFAs and lactic acid are reviewed . Analysis of the organic acids involved in carbohydrate metabolism has been achieved by enzymatic methods, gas chromatography, high-performance liquid chromatography and capillary electrophoresis . Sample preparation techniques developed for these analytes are also discussed.

Biosci Biotechnol Biochem, 2002 Oct, 66(10), 2216 - 20
RNase G-dependent degradation of the eno mRNA encoding a glycolysis enzyme enolase in Escherichia coli; Kaga N et al.; Escherichia coli RNase G, encoded by the rng gene, is involved in the processing of 16S rRNA and degradation of the adhE mRNA encoding a fermentative alcohol dehydrogenase . In a search for the intracellular target RNAs of RNase G other than the 16S rRNA precursor and adhE mRNA, total cellular proteins from rng+ and rng::cat cells were compared by two-dimensional gel electrophoresis . The amount of enolase encoded by the eno gene reproducibly increased two- to three-fold in the rng::cat mutant strain compared with the rng+ parent strain . Rifampicin chase experiments showed that the half-life of the eno mRNA was some 3 times longer in the rng::cat mutant than in the wild type . These results indicate that the eno mRNA was a substrate of RNase G in vivo, in addition to 16S rRNA precursor and adhE mRNA.

Prikl Biokhim Mikrobiol, 2002 Nov-Dec, 38(6), 639 - 43
Production, characterization and properties of beta-glucosidase and beta-xylosidase from a strain of Aureobasidium sp; Iembo T et al.; beta-glucosidase and beta-xylosidase production by a yeast-like Aureobasidium sp . was carried out during solid-state and submerged fermentation using different carbon sources and crude enzymes were characterized . beta-glucosidase and beta-xylosidase gave optimum activity at pH 2.0-2.5 and 3.0, respectively . These enzymes presented maximal activity at 65 degrees, and were stable in a wide pH range and high temperatures.

Water Res, 2002 Dec, 36(20), 4985 - 96
Comparison of trichloroethylene reductive dehalogenation by microbial communities stimulated on silicon-based organic compounds as slow-release anaerobic substrates; Yu S et al.; Microcosm studies were conducted to demonstrate the effectiveness of tetrabutoxysilane (TBOS) as a slow-release anaerobic substrate to promote reductive dehalogenation of trichloroethylene (TCE) . The abiotic hydrolysis of TBOS and tetrakis(2-ethylbutoxy)silane (TKEBS), and the biotic transformations of the hydrolysis products from both were also investigated . Comparison of TCE reductive dehalogenation was performed with microbial communities stimulated from three different sites: Site 300 Lawrence Livermore National Laboratory (LLNL), CA, Point Mugu Naval Weapons Facility, CA, and the Evanite site in Corvallis, OR . Poisoned microcosms showed that 1 mol of TBOS slowly and abiotically hydrolyzes to 4 mol of 1-butanol, while the live microcosms showed the 1-butanol ferments to butyrate and/or acetate, producing H2 . The hydrolysis of TBOS and TKEBS was abiotic and not enhanced by biotic processes under the anaerobic conditions of these tests . Hydrogen consumption was correlated with reductive dehalogenation, indicating it served as an electron donor for reductive dehalogenation . TBOS was found to be a slow-release anaerobic substrate to support long-term dechlorination of TCE to ethylene in Point Mugu microcosms, and in the LLNL microcosm bioaugmented with the Evanite culture . Electron mass balances showed most of the electron flow went into the creation of organic acids, especially acetate, and the production of methane . Electron efficiencies for reductive dechlorination were as high as 14% based on the electrons used for dechlorination to the total electrons associated with the mass of TBOS and TKEBS hydrolyzed . Rates of TBOS hydrolysis increased with greater TBOS concentrations as a light nonaqueous-phase liquids (LNAPL) . These results indicate that TBOS has promise as an effective anaerobic substrate for remediating a wide range of CAH concentrations at different CAH contaminated sites.

J Bacteriol, 2002 Dec, 184(24), 6929 - 41
Molecular genetic analysis of ICEF, an integrative conjugal element that is present as a repetitive sequence in the chromosome of Mycoplasma fermentans PG18; Calcutt MJ et al.; Mycoplasma genomes contain compact gene sets that approach the minimal complement necessary for life and reflect multiple evolutionary instances of genomic reduction . Lateral gene transfer may play a critical role in shaping the mobile gene pool in these organisms, yet complex mobile elements have not been reported within this genus . We describe here a large ( approximately 23-kb) genetic element with unique features that is present in four copies in the Mycoplasma fermentans PG18 chromosome, accounting for approximately 8% of the genome . These novel elements, designated ICEF (integrative conjugal elements of M . fermentans), resemble conjugative, self-transmissible integrating elements (constins) in that circular, nonreplicative extrachromosomal forms occur in which the left and right termini of the integrated element are juxtaposed and separated by a coupling sequence derived from direct repeats flanking chromosomal copies of ICEF as a result of target site duplication . ICEF contain multiple similarly oriented open reading frames (ORFs), of which some have homology to products of known conjugation genes but others have no known counterparts . Surprisingly, unlike other constins, ICEF lack homologs of known integrases, transposases, or recombinases, suggesting that a novel enzyme may be employed for integration-excision . Skewed distribution and varied sites of chromosomal integration among M . fermentans isolates suggest a role for ICEF in promoting genomic and phenotypic variation in this species . Identification of homologs of terminal ICEF ORFs in two additional mycoplasma species indicates that ICEF is the prototype member of a family of ICE-related elements that may be widespread among pathogenic mycoplasmas infecting diverse vertebrate hosts.

J Nat Prod, 2002 Nov, 65(11), 1734 - 7
Bioactive natural products from a sclerotium-colonizing isolate of Humicola fuscoatra; Joshi BK et al.; Chemical studies of an organic extract from solid-substrate fermentations of the mycoparasitic fungus Humicola fuscoatra NRRL 22980, originally isolated as a colonist of Aspergillus flavus sclerotia, afforded two new unrelated compounds that we have named fuscoatroside (1) and fuscoatramide (2) . The structures of these metabolites were elucidated by analysis of NMR and MS data . Fuscoatroside (1) is a triterpenoid glycoside that exhibited activity in antifungal assays against A . flavus . This extract also contained the known metabolites 7-deoxysterigmatocystin, sterigmatocystin, isosclerone, and decarestrictines A(1) and I.

J Nat Prod, 2002 Nov, 65(11), 1582 - 7
Accumulation of epigallocatechin quinone dimers during tea fermentation and formation of theasinensins; Tanaka T et al.; Production and accumulation of catechin dimer quinones during tea fermentation were chemically confirmed for the first time by trapping as phenazine derivatives . Direct treatment of the fermented tea leaves with o-phenylenediamine yielded five phenazine derivatives (8-12) of o-quinones of an epigallocatechin dimer and its galloyl esters (13-16), in which two flavan units were linked at the B-rings through a C-C bond . Atrop isomerism of the biphenyl bonds was shown to be the R configuration, suggesting that the o-quinone dimers were generated by stereoselective coupling of monomeric quinones . The total concentration of the phenazine derivatives in the o-phenylenediamine-treated tea leaves was higher than that of theaflavins . In contrast, phenazine derivatives of monomeric quinones of epigallocatechin were not isolated . When the fermented tea leaves were heated, the quinone dimers were converted to theasinensins, which are constituents of black tea, suggesting that theasinensins are generated by reduction of the quinone dimers during the heating and drying steps in black tea manufacturing.

Trends Biotechnol, 2002 Dec, 20(12), 515 - 21
The production of biocatalysts and biomolecules from extremophiles; Schiraldi C et al.; The discovery of life in seemingly prohibitive environments continues to challenge conventional concepts of the growth-limiting conditions of many cellular organisms . The diversity of extremophiles has barely been tapped -estimates generally agreeing that <1% of the microorganisms in the environment have been cultivated in pure cultures to date . The production of extremophilic biomass is very important to provide sufficient material for enzyme and biomolecule isolation and characterization, eventually revealing particular features of industrial interest . Hence, special equipment and custom-tailored processes have been developed and are currently under evaluation for the improvement of fermentation productivity . Despite the remarkable opportunities that these uncommon organisms present for biotechnological applications only few instances can be reported for actual exploitation . This lack of progress from the research findings at a laboratory-scale to the actual development of pilot and large-scale production is correlated with the difficulties encountered in extremophile cultivations . Here, we report recent achievements in the production of biomass and related enzymes and biomolecules from extremophile sources, especially focusing on the application of novel fermentation strategies.

J Environ Radioact, 2002, 63(3), 265 - 70
The association of 137Cs with various components of tea leaves fermented from Chernobyl contaminated green tea; Polar E; The distribution of 137Cs among various components of fermented tea leaves harvested after the Chernobyl accident was investigated by applying a sequential extraction procedure . An association of the radioisotope with the phenolic moiety of a phenylglucoside was detected in black tea infusate using permeation chromatography as well as UV and NMR spectroscopy . The chemical structure of a 137Cs containing compound also isolated from an artificially 137Cs labelled aqueous extract of green leaves was compared to that of fermented tea leaves . The implications of the findings on the chemical forms of the radionuclide are discussed.

Transplantation, 2002 Oct 27, 74(8), 1200 - 1
Rhabdomyolysis due to red yeast rice (Monascus purpureus) in a renal transplant recipient; Prasad GV et al.; Rhabdomyolysis is a known complication of hepatic 3-methylglutaryl coenzyme A reductase (HMG-CoA) inhibitor (statin) therapy for posttransplant hyperlipidemia, and thus monitoring for this effect is indicated . We report a case of an herbal preparation-induced rhabdomyolysis in a stable renal-transplant recipient, attributed to the presence of red yeast rice (Monascus purpureus) within the mixture . The condition resolved when consumption of the product ceased . Rice fermented with red yeast contains several types of mevinic acids, including monacolin K, which is identical to lovastatin . We postulate that the interaction of cyclosporine and these compounds through the cytochrome P450 system resulted in the adverse effect seen in this patient . Transplant recipients must be cautioned against using herbal preparations to lower their lipid levels to prevent such complications from occurring.

New Microbiol, 2002 Oct, 25(4), 477 - 84
Ester antibiotic accumulation by Streptomyces hygroscopicus; Bhattacharyya BK et al.; In an attempt to maximize the ester antibiotic production by Streptomyces hygroscopicus D1.5, its efficacy was found to be enhanced by manipulation of the nutrient and physical environment . The two stage fermentation using seed inoculum (10% v/v) resulted in better production while fermentation continued for 5 days in pH 7.0 at 30 degrees C . Enhanced yield was also observed in whole cell immobilization . Under entrapment, maximum yield was achieved at 7th and 9th day of fermentation for mycelia and spore . In addition, the beads could be reused up to the 3rd cycle.

Biotechnol Annu Rev, 2002, 8, 183 - 225
Overview of solid state bioprocessing; Mitchell DA et al.; Solid-state fermentation has centuries of history, but it is only in the last two decades that there has been a concerted effort to understand the bioprocessing issues involved and to apply them to a wide range of new products . This article provides an overview of the knowledge of solid-state bioprocessing that has been gained over this time . It shows that, although significant advances have been achieved in understanding of what controls process performance, much research is still required.

Appl Microbiol Biotechnol, 2002 Nov, 60(3), 281 - 7 Epub 2002 Oct 05.
Production of 5,8,11-eicosatrienoic acid by a delta5 and delta6 desaturation activity-enhanced mutant derived from a delta12 desaturation activity-defective mutant of Mortierella alpina 1S-4; Sakuradani E et al.; Enhanced production of 5,8,11-eicosatrienoic acid (Mead acid, 20:3omega9) was attained with a mutant fungus, Mortierella alpina JT-180, derived from delta12 desaturation activity-defective and delta6 desaturation activity-enhanced M . alpina M209-7 . Production of 20:3omega9 by JT-180 was 1.4 times greater than that of the parent strain M209-7 . This is thought to be due to its enhanced Delta5 desaturation activity, which was 3.3 times higher than that of M209-7 . In both strains, 78.5-80.4% of the total lipids comprised triacylglycerol (TG), and 76.6-79.0% of 20:3omega9 was present in TG . Comparing the fatty acid compositions among various lipid species, the highest percentages (24.1-37.6%) of 20:3omega9 in total lipids were found in phosphatidylcholine . For optimization of 20:3omega9 production by JT-180, a glucose concentration of 4% in the culture medium and shifting of the growth temperature from 28 degrees C to 20 degrees C on the 2nd day were shown to be effective . Under optimal conditions, 20:3omega9 production by JT-180 reached 1.92 g/l culture medium in a 10-l jar fermentor (corresponding to 81.5 mg/g dry mycelia and 18.3% of total fatty acids), which is greater than that reported previously from M209-7 (1.65 g/l).

Appl Microbiol Biotechnol, 2002 Nov, 60(3), 233 - 42 Epub 2002 Oct 12.
Biochemistry and biotechnological applications of Gluconobacter strains; Deppenmeier U et al.; The genus Gluconobacter belongs to the group of acetic acid bacteria, which are characterized by their ability to incompletely oxidize a wide range of carbohydrates and alcohols . The corresponding products (aldehydes, ketones and organic acids) are excreted almost completely into the medium . In most cases, the reactions are catalyzed by dehydrogenases connected to the respiratory chain . Since the reactive centers of the enzymes are oriented towards the periplasmic space, transport of substrates and products into, and out of, the cell is not necessary . Thus, rapid accumulation of incompletely oxidized products in the medium is facilitated . These organisms are able to grow in highly concentrated sugar solutions and at low pH-values . High oxidation rates correlate with low biomass production, which makes Gluconobacter strains interesting organisms for industrial applications . Modern fermentation processes, such as the production of L-sorbose (vitamin C synthesis) and 6-amino- L-sorbose (synthesis of the antidiabetic drug miglitol) are carried out with members of this genus . Other important products are dihydroxyacetone, gluconate and ketogluconates . The bacteria belonging to the genus Gluconobacter exhibit extraordinary uniqueness not only in their biochemistry but also in their growth behavior and response to extreme culture conditions . This uniqueness makes them ideal organisms for microbial process development.

Nat Prod Lett, 2002 Oct, 16(5), 345 - 9
Microbial transformation of (+)-adrenosterone; Musharraf SG et al.; The microbial transformation of (+)-adrenosterone (1) by Cephalosporium aphidicola afforded three metabolites identified as androsta-1,4-diene-3,11,17-trione (2), 17beta-hydroxyandrost-4-ene-3,11-dione (3) and 17beta-hydroxyandrosta-1,4-diene-3,11-dione (4) . The fermentation of 1 with Fusarium lini also produced metabolites 2 and 4, while the fermentation with Trichothecium roseum afforded metabolite 3 . The structures of transformed products were determined by spectroscopic methods.

Anal Bioanal Chem, 2002 Nov, 374(5), 835 - 40 Epub 2002 Oct 08.
Separation and identification of organic acid-coenzyme A thioesters using liquid chromatography/electrospray ionization-mass spectrometry; Dalluge JJ et al.; A method has been developed for the direct determination of coenzyme A (CoA) and organic acid-CoA thioesters in mixtures using directly combined liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS) . Mixtures of CoA and organic acid-CoA thioesters were analyzed by LC/ESI-MS with detection of protonated molecular ions and characteristic fragment ions for each compound . The identities of the CoA-thioesters were established based on LC retention times and simultaneously recorded mass spectra . Monitoring of the CoA specific fragment ion at m/z 428 throughout the chromatogram provides a unique fingerprint for CoA content in the samples that corroborates the identification of organic acid-CoA thioesters in the mixtures . Furthermore, fragment ions arising from the ester linkage portion of the molecule allow unambiguous identification of the CoA esters in the samples . A second LC elution system was developed that allows the simultaneous separation and identification of 2-hydroxypropionyl-CoA (lactyl-CoA) and 3-hydroxypropionyl CoA (3HP-CoA), which have the same mass and identical MS fragmentation behavior . The utility of LC/ESI-MS employing this elution system is demonstrated by the determination of 3HP-CoA and lactyl-CoA (converted to CoA-thioesters from their corresponding free acids using CoA-transferase) in fermentation broths from Escherichia coli strains engineered for the production of 3-hydroxypropionic acid (3HP) . External calibration employing a purified 3HP-CoA standard allowed indirect quantification of 3HP content in the broth with a precision of 1% (RSD) . The feasibility of extending the method described above to perform LC/selected reaction monitoring-tandem mass spectrometry for direct determination of organic acid-CoA thioesters in cells was also demonstrated.

Curr Microbiol, 2003 Jan, 46(1), 24 - 7
Effects of carboxymethylcellulose and carboxypolymethylene on morphology of Aspergillus fumigatus NRRL 2346 and fumagillin production; Yang W et al.; Aspergillus fumigatus NRRL 2346 is the producer of fumagillin, an antitumor antibiotic that inhibits angiogenesis . This strain is very difficult to grow reproducibly in shake flasks owing to an extreme form of pellet growth and extensive wall growth . The effects of carboxymethylcellulose (CMC) and carboxypolymethylene (Carbopol) on growth and fumagillin production by A . fumigatus were investigated . By adding the polymers to the fermentation medium, the growth form of the mold was changed from a single large glob to small reproducible pellets, and wall growth was diminished to a minimum . Carbopol, at a lower concentration, was more effective than CMC in improving both morphology and production . Small pellets were produced which favored fumagillin biosynthesis . 1.5% (wt/vol) CMC and 0.3% (wt/vol) Carbopol were found to be the optimum concentrations; higher levels increased viscosity to an unacceptable level.

Int J Food Microbiol, 2003 Jan 15, 80(1), 61 - 6
Solid-state fermentation of cornmeal with the basidiomycete Hericium erinaceum for degrading starch and upgrading nutritional value; Han J; The ability of the basidiomycete Hericium erinaceum to degrade starch and upgrade nutritional value of cornmeal during solid-state fermentation was studied . On the basal medium which consisted of cornmeal and salt solution, H . erinaceum produced a strong alpha-amylase on the 15th day after inoculation, which resulted in a 52% degradation of the starch . By supplementation with 5-15 g soybean meal per 100 g cornmeal the alpha-amylase activity and degradation rate of starch was raised significantly (P < 0.01) . Prolongation of fermentation time from 15 to 30 days did not increase significantly the degradation rate of starch, though the alpha-amylase activity reached its maximum value of 179 U/g on the 20th day after inoculation . Under conditions close to the theoretical optimum fermentation conditions, that was after 25 days at 25 degrees C in the medium with added 15 g soybean meal per 100 g cornmeal, the starch content in the product decreased from 63% to 22% (P < 0.001) and protein content increased from 12% to 17% (P < 0.01) . In the protein in the product, the lysine content was increased from 36 to 56 mg/ g and tryptophan from 9 to 13 mg/g . Using egg protein as a standard, an evaluation on the protein quality of the fermented product showed that it was superior to that of the nonfermented control and to other cereals, was close to that of soybean and chicken, but was inferior to that of milk and red meats.

Int J Food Microbiol, 2003 Jan 15, 80(1), 47 - 53
Effects of fermentation temperature on the strain population of Saccharomyces cerevisiae; Torija MJ et al.; The influence of fermentation temperature (from 15 to 35 degrees C) on a mixed strain population was studied . Mitochondrial DNA analysis was used to differentiate Saccharomyces cerevisiae strains and the frequency of each strain during the alcoholic fermentation was determined . The chemical analyses of resulting wines were carried out . The temperature determined how Saccharomyces strains developed and how effectively they fermented . Some strains performed better at high temperatures and others at low temperatures . The maximal population size was similar at all temperatures . At low temperatures, however, it was reached later though it remained constant throughout the alcoholic fermentation . On the other hand, viable cells decreased at high temperatures, especially at 35 degrees C . Obviously, the composition of the wines changed as the temperature of fermentation changed . At low temperatures, alcohol yield was higher . Secondary metabolites to alcoholic fermentation increased as the temperature increased . Glycerol levels were directly affected by temperature.

J Agric Food Chem, 2002 Nov 20, 50(24), 7042 - 5
Influence of fatty acids on wine foaming; Gallart M et al.; The influence of fatty acids (free and bound as ethyl esters) on wine foaming was studied in different white wines and the corresponding sparkling wines . Moreover, from three of these wines the foam formed by CO(2) injection was separated, and two fractions were then obtained: foam wine (FW) and remainder wine (RW) . In these fractions and the sparkling wines produced from them, foam properties and fatty acids were also determined . The free fatty acids C8, C10, and C12 were negatively correlated with foamability (HM), whereas the ethyl esters of hexanoic, octanoic, and decanoic acids were positively related to HM . The value of HM was directly proportional to the ratio of esterified to unesterified fatty acids . This was confirmed by the changes that occur in the esterification ratio during the second fermentation and aging . No influence was observed on either the Bikerman coefficient or the stability time of foam.

OMICS, 2002, 6(3), 247 - 58
Suspected utility of enzymes with multiple activities in the small genome Mycoplasma species: the replacement of the missing "household" nucleoside diphosphate kinase gene and activity by glycolytic kinases; Pollack JD et al.; The small genome Mollicutes whose DNAs are completely sequenced (Mycoplasma genitalium, Mycoplasma pneumoniae, Mycoplasma pulmonis, and Ureaplasma urealyticum {parvum}) lack a gene (ndk) for the presumably essential nucleoside diphosphate kinase (NDPK) . We hypothesized that other activities might replace NDPK activity . We found in M . genitalium G37(T), Mycoplasma pneumoniae FH(T), Mycoplasma fermentans PG18(T), and Mycoplasma capricolum subsp . capricolum Kid(T) that their 6-phosphofructokinases (6-PFKs), phosphoglycerate kinases (PGKs), pyruvate kinases (PKs), and acetate kinases (AKs), besides reactant ADP/ATP, could use other ribo- and deoxyribo-purine and pyrimidine NDPs and NTPs . These activities could compensate for the absence of an orthologous ndk gene in the Mycoplasmataceae . They suggest a metabolically varied and consequential role for unrelated and perhaps unsuspected "replacement" or compensatory enzymes that may confound metabolic prediction . We partially purified and biochemically characterized the PKs, 6-PFKs, PGKs, and AKs from M . capricolum subsp . capricolum Kid(T) and M . fermentans PG18(T).

J Bacteriol, 2002 Dec, 184(23), 6642 - 53
Regulation of the hydrogenase-4 operon of Escherichia coli by the sigma(54)-dependent transcriptional activators FhlA and HyfR; Skibinski DA et al.; The hyf locus (hyfABCDEFGHIJ-hyfR-focB) of Escherichia coli encodes a putative 10-subunit hydrogenase complex (hydrogenase-4 {Hyf}); a potential sigma(54)-dependent transcriptional activator, HyfR (related to FhlA); and a putative formate transporter, FocB (related to FocA) . In order to gain insight into the physiological role of the Hyf system, we investigated hyf expression by using a hyfA-lacZ transcriptional fusion . This work revealed that hyf is induced under fermentative conditions by formate at a low pH and in an FhlA-dependent fashion . Expression was sigma(54) dependent and was inhibited by HycA, the negative transcriptional regulator of the formate regulon . Thus, hyf expression resembles that of the hyc operon . Primer extension analysis identified a transcriptional start site 30 bp upstream of the hyfA structural gene, with appropriately located -24 and -12 boxes indicative of a sigma(54)-dependent promoter . No reverse transcriptase PCR product could be detected for hyfJ-hyfR, suggesting that hyfR-focB may be independently transcribed from the rest of the hyf operon . Expression of hyf was strongly induced ( approximately 1,000-fold) in the presence of a multicopy plasmid expressing hyfR from a heterologous promoter . This induction was dependent on low pH, anaerobiosis, and postexponential growth and was weakly enhanced by formate . The hyfR-expressing plasmid increased fdhF-lacZ transcription just twofold but did not influence the expression of hycB-lacZ . Interestingly, inactivation of the chromosomal hyfR gene had no effect on hyfA-lacZ expression . Purified HyfR was found to specifically interact with the hyf promoter/operator region . Inactivation of the hyf operon had no discernible effect on growth under the range of conditions tested . No Hyf-derived hydrogenase or formate dehydrogenase activity could be detected, and no Ni-containing protein corresponding to HyfG was observed.

Br J Nutr, 2002 Nov, 88(5), 523 - 32
Increasing the viscosity of the intestinal contents stimulates proliferation of enterotoxigenic Escherichia coli and Brachyspira pilosicoli in weaner pigs; Hopwood DE et al.; The present study was designed to evaluate the effect of increased viscosity of the intestinal digesta on proliferation of enterotoxigenic Escherichia coli and the intestinal spirochaete Brachyspira pilosicoli in weaned pigs . Pigs were fed an experimental diet based on cooked white rice (R), which was supplemented with carboxymethylcellulose (CMC; 40 g/kg diet) to increase digesta viscosity . Thirty-six piglets weaned at 21 d of age were divided into six groups, three of which were fed R and three Addition of CMC increased digesta viscosity in the ileum (P=0.01), caecum (P=0.0007) and colon (P=0.0035), without increasing indices of large intestinal fermentation . Pigs fed developed a natural infection with enterotoxigenic E . coli after weaning and had more (P<0.0001) diarrhoea than pigs fed R . Subsequent experimental infection of two groups of pigs with B . pilosicoli resulted in more (P<0.0001) colonisation in pigs fed than R . At this time, all pigs fed had wetter (P<0.0001) faeces than those fed R, irrespective of whether they were infected with B . pilosicoli, but infected pigs also had an increased (P=0.025) number of days with diarrhoea post-infection irrespective of diet . In pigs fed it was not clear to what extent the increased viscosity associated with CMC, or the concurrent infection with enterotoxigenic E . coli, was responsible for the increased proliferation of B . pilosicoli . In a second experiment, five pigs that were weaned onto an R diet were transferred onto 3 weeks later . These pigs did not develop a natural infection with enterotoxigenic E . coli after the diet change, confirming the particular susceptibility of pigs to enterotoxigenic E . coli proliferation immediately post-weaning.

Br J Nutr, 2002 Nov, 88(5), 489 - 98
Large bowel fermentation of maize or sorghum-acorn diets fed as a different source of carbohydrates to Landrace and Iberian pigs; Morales J et al.; Twenty-four finishing pigs (twelve Iberian and twelve Landrace) were used in a growing and slaughtering experiment . Animals were fed two diets differing in their ingredients, maize (diet C) or sorghum-acorn (diet A) . At an average weight of 107.0 kg pigs were slaughtered and hindgut digesta sampled to study the effect of breed and diet on large bowel fermentation . Flows of digesta to the hindgut compartment were estimated based on an indigestible flow marker (Cr2O3) and were higher in Iberian than in Landrace pigs (P<0.001), and higher in animals fed diet A than diet C (P=0.07) . The higher flows in Iberian pigs were mainly associated with a higher voluntary feed intake (3.50 v . 2.70 kg/d, P<0.01) and lower ileal digestibility of NSP (-12.8 v . 47.8, P<0.01) . Differences between diets were mainly associated with a lower ileal digestibility of starch from diet A (89.2 v . 96.9 %, P=0.06), although no differences in the resistant starch content were observed in vitro . Fermentation of different carbohydrates through the large bowel showed that NSP-glucose had lower digestibility in Iberian than in Landrace pigs (62.5 v . 94.2 %, P<0.001), but no differences were observed in starch, or other NSP-fibre fractions (arabinose, xylose and galactose) . The type and amount of carbohydrates reaching the large bowel were related to the diet but also to breed, and promoted differences in the fermentative activity associated with different volatile fatty acid patterns and changes in microbial enzymic activity.

Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet, 2001, 66(2a), 375 - 86
Spinosad, a new tool for insect control in vegetables cultivated in greenhouses; Schoonejans T et al.; Spinosad is a biological insecticide derived as a fermentation product from the soil actinomycete Saccharopolyspora spinosa . The compound was tested for its possible use in northwestern Europe in vegetables cultivated in greenhouses . Spinosad is an insect control agent that has activity against a wide range of insect pests including caterpillars, leafminers and thrips . Because of its selectivity on a large number of beneficials used in greenhouses, spinosad is an interesting tool for insect control . A SC formulated product, containing 480 g a.i . litre-1, was tested in 1998, 1999 and 2000 in a range of trials at the research stations of Naaldwijk/Netherlands, and Gembloux/Belgium . This paper reviews and discusses the efficacy results on Trialeurodes vaporarium, Frankliniella occidentalis, Chrysodeixis Chalcites, Liriomyza Bryoniae . According the target insect rates from 4.8 till 36 g a.i./hl were tested . Consistent performance was recorded on F . occidentalis at rates of 9.6 g a.i./hl or above . Interesting efficacy results were recorded on the other above-mentioned insects.

Int J Food Microbiol, 2003 Feb 25, 81(1), 63 - 71
A new PCR-based method for monitoring inoculated wine fermentations; Lopez V et al.; A new PCR-based method has been developed to monitor inoculated wine fermentations . The method is based on the variation in the number and position of introns in the mitochondrial gene COX1 . Oligonucleotide primers homologous to the regions flanking the Saccharomyces cerevisiae COX1 introns have been designed and tested for S . cerevisiae wine yeast strain differentiation . Four primers were selected for their subsequent use in a multiplex PCR reaction and have proved to be very effective in uncovering polymorphism in natural and commercial yeast strains . An important point is that the speed and simplicity of the technique, which does not require the isolation of DNA, allows early detection of the starter yeast strain throughout the fermentation process . The main advantage for the wineries is that the must sample can be used directly for the PCR reaction obtaining very fast results (in approximately 8 h) . This allows the wine industries to intervene quickly if necessary.

Phytochemistry, 2002 Nov, 61(6), 721 - 6
Composition of sugarcane waxes in rum factory wastes; Nuissier G et al.; Wastes produced during fermentation and distillation of crude sugarcane juice in rum factories were evaluated as a new source of waxes . The chemical composition of the crude wax extracted from adsorbat of the wastes on fuller's earth was studied by GC-mass spectrometry . Series of linear alkanes (C19-C33), and wax esters constitute the main components . In addition, phytosterols, triterpene methyl ethers, ethyl and methyl esters of fatty acids, and free fatty acids were found as minor components . Acid (predominance of C16 and C18) and alcohol portions (C26-C32) of the wax esters were analysed after saponification.

FEMS Immunol Med Microbiol, 2002 Nov 15, 34(3), 209 - 14
High prevalence of Mycoplasma infections among European chronic fatigue syndrome patients . Examination of four Mycoplasma species in blood of chronic fatigue syndrome patients; Nijs J et al.; Prevalence of Mycoplasma species infections in chronic fatigue syndrome (CFS) has been extensively reported in the scientific literature . However, all previous reports highlighted the presence of Mycoplasmas in American patients . In this prospective study, the presence of Mycoplasma fermentans, M . penetrans, M . pneumoniae and M . hominis in the blood of 261 European CFS patients and 36 healthy volunteers was examined using forensic polymerase chain reaction . One hundred and seventy-nine (68.6%) patients were infected by at least one species of Mycoplasma, compared to two out of 36 (5.6%) in the control sample (P<0.001) . Among Mycoplasma-infected patients, M . hominis was the most frequently observed infection (n=96; 36.8% of the overall sample), followed by M . pneumoniae and M . fermentans infections (equal frequencies; n=67; 25.7%) . M . penetrans infections were not found . Multiple mycoplasmal infections were detected in 45 patients (17.2%) . Compared to American CFS patients (M . pneumoniae>M . hominis>M . penetrans), a slightly different pattern of mycoplasmal infections was found in European CFS patients (M . hominis>M . pneumoniae, M . fermentansz.Gt;M . penetrans).

Folia Microbiol (Praha), 2002, 47(4), 407 - 8
Winemaking from Gaglioppo grapes with hybrid strains of Saccharomyces; Caridi A et al.; The fermentative behavior of two hybrid wine yeast strains (first-generation hybrid-strain 12,233 x 6167--obtained by hybridization of the cryotolerant strain S . bayanus 12,233 with the mesophilic strains S . cerevisiae 6167, and TT254 x 6392 arising by hybridization of the thermotolerant strain S . cerevisiae TT254 with the mesophilic strain S . cerevisiae 6392) was compared with that of a commercial wine yeast strain S . cerevisiae K1 in must from black grapes of the Calabrian variety Gaglioppo . The goal was to obtain wines with a high content of 'polyphenols' from a grape must with a limited phenolic content such as the Gaglioppo must . The progress of the winemaking was estimated according to residual sugars; at the end of fermentation, the wines were decanted, bottled and principal physico-chemical characteristics determined . Our results point to the possibility to select wine yeasts (significantly differing in the above parameters) by their ability to interact with phenolic compounds.

Folia Microbiol (Praha), 2002, 47(4), 385 - 90
Variability of laccase activity in the white-rot basidiomycete Pleurotus ostreatus; Baldrian P et al.; The production of laccase in liquid cultures of the white-rot fungus Pleurotus ostreatus was highly variable . During the first days of cultivation, the relative variability was as high as 80-100% and it decreased to 30% in the course of cultivation . The main source of variability was assumed to be the independent development of enzyme activity in individual cultures . Cultures with high laccase production showed also high production of the other ligninolytic enzyme--Mn-dependent peroxidase . The variability was probably due to the source of inoculum, deactivation of the enzyme in culture liquid and genetic variations among the cultures . Variability of laccase activities was lower during solid-state fermentation on wheat straw and during the growth in nonsterile soil.

Bioresour Technol, 2003 Jan, 86(1), 85 - 9
Mesophilic biogas production from fruit and vegetable waste in a tubular digester; Bouallagui H et al.; A semi-continuously mixed mesophilic tubular anaerobic digester was tested for the conversion of fruit and vegetable waste (FVW) into biogas . The effect of hydraulic retention time (HRT) and the feed concentration on the extent of the degradation of the waste was examined . Varying the HRT between 12 and 20 days had no effect on the fermentation stability and pH remained between 6.8 and 7.6, but an inhibition of methanogenic bacteria was observed at HRT below 12 days . The overall performance of the reactor was depressed by changing the feed concentration from 8% to 10% TS (dry weight) . By applying a feed concentration of 6% and HRT of 20 days in the tubular digester, 75% conversion efficiency of FVW into biogas with a methane content of 64% was achieved.

Bioresour Technol, 2003 Jan, 86(1), 29 - 32
Utilization of fermented silkworm pupae silage in feed for carps; Rangacharyulu PV et al.; Fermented silkworm pupae (SWP) silage or untreated fresh SWP pastes were incorporated in carp feed formulations replacing fishmeal . The feed formulations were isonitrogenous (30.2-30.9% protein) and isocaloric (ME = 2905-2935 kcal/kg) . Feeding under a polyculture system consisting of 30% each of catla (Catla catla), mrigal (Cirrhinus mrigala) and rohu (Labeo rohita) with 10% silver carps (Hypophthalmychthys molitrix) was carried out in ponds to evaluate the nutritive quality of SWP silage . Survival rate, feed conversion ratio and specific growth rate, respectively, were 84.2%, 2.10 and 2.39 for fermented SWP silage, 65.8%, 2.98 and 2.26 for untreated SWP and 67.5%, 3.16 and 2.20 for fishmeal indicating clearly that the fermented SWP silage was nutritionally superior to untreated SWP or fishmeal . The dietary influence on the proximate composition of whole fish was marginal.

Eur J Nutr, 2002 Nov, 41 Suppl 1, I2 - 10
The physiology of colonic metabolism . Possibilities for interventions with pre- and probiotics; Priebe MG et al.; The awareness is increasing that in the colon many metabolic processes take place in relation to the fermentation of our food, which might be relevant for health and disease . However, the relation between food, colon metabolism and health or disease is far from clear . In this overview, the physiology of colonic metabolism and possibilities for its modification by the use of pre- and probiotics are discussed . Results of in vitro and animal studies indicate a beneficial impact of probiotics on adverse metabolic processes in the colon, but confirmation in human studies has to be extended . The administration of prebiotics seems to be promising with regard to their capacity to modulate the bacterial composition in the colon and there are indications that prebiotics can beneficially influence colonic metabolism . Whether these modulations brought about by pre- or probiotics have an effect on the health of the host, however, needs to be established in most cases.

Chem Pharm Bull (Tokyo), 2002 Nov, 50(11), 1427 - 9
Microbial transformation of rubijervine; El Sayed KA et al.; Preparative-scale fermentation of rubijervine (1), the known 22,26-epiminocholestane Veratrum alkaloid, with Cunninghamella echinulata ATCC 9244 has resulted in the isolation of the new metabolites 7alpha-hydroxyrubijervine (2) and solanid-5-ene-3beta,12alpha-diol-1-one (3) . Structure elucidation of these metabolites was based primarily on 1D- and 2D-NMR analyses . The microbe C . echinulata ATCC 9244 was able to metabolize rings A and B of rubijervine but failed to metabolize rings C, D or its N-containing side chain, a finding which is analogous to the results of previous fermentation studies of steroidal alkaloids.

Anal Biochem, 2002 Sep 15, 308(2), 204 - 9
Direct determination of tryptophan using high-performance anion-exchange chromatography with integrated pulsed amperometric detection; Hanko VP et al.; Here we present a new method to rapidly quantify tryptophan (Trp) in proteins, animal feed (Mehaden fishmeal), cell cultures, and fermentation broths . Trp is separated from common amino acids by anion-exchange chromatography in 12min and directly detected by integrated pulsed amperometry . The estimated lower detection limit for this method is 1pmol . Alkaline (4M NaOH) hydrolysates can be directly injected, and therefore we used this method to determine the optimum alkaline hydrolysis conditions for the release of Trp from a model protein, bovine serum albumin (BSA) . This method accurately determined the Trp content of BSA and fishmeal . High levels of glucose (2%, w/w) do not interfere with the chromatography or decrease recovery of Trp . We used this method to monitor free Trp during an Escherichia coli fermentation.

Nutr Health, 2002, 16(3), 173 - 81
Effect of probiotic fermentation on antinutrients and in vitro protein and starch digestibilities of indigenously developed RWGT food mixture; Sindhu SC et al.; Indigenously developed RWGT food mixture which contained rice flour, whey, sprouted green gram paste and tomato pulp (2:1:1:1 w/w) was autoclaved 1.5 kg/cm2, 15 min, 121 degrees C), cooled and fermented with 2% liquid culture (containing 10(6) cells/ml broth) . Two types of fermentations were carried out i.e . single culture fermentation {L . casei, L . plantarum (37 degrees C, 24 hr.)} and sequential culture fermentation {S . boulardii (25 degrees C, 24 hr.) + L . casei (37 degrees C, 24 hr.)}; S . boulardii {(25 degrees C, 24 hr.) + L . plantarum (37 degrees C, 24 hr.)} . All the fermentations drastically reduced the contents of phytic acid, polyphenols and trypsin inhibitor activity while significantly improving the in vitro digestibilities of starch and protein . Sequential culture fermentations brought about higher changes as compared to single culture fermentations.

Trends Plant Sci, 2002 Nov, 7(11), 478 - 81
Function of the alternative oxidase: is it still a scavenger?
Moore AL, Albury MS, Crichton PG, Affourtit C.
The alternative oxidase is a respiratory chain protein found in all higher plants, fungi, non-fermentative yeasts and trypanosomes . Its primary structure suggests that it is a new member of the di-iron carboxylate protein family . Recent sequence analysis indicates an evolutionary relationship between primitive members of this protein family and the alternative oxidase, suggesting that its early function was to scavenge di-oxygen . However, modelling of plant growth kinetics suggests a different function.

J Dairy Sci, 2002 Oct, 85(10), 2603 - 8
Effects of Saccharomyces cerevisiae culture and Saccharomyces cerevisiae live cells on in vitro mixed ruminal microorganism fermentation; Lynch HA et al.; The objective of this study was to examine the effects of a Saccharomyces cerevisiae live cell product and a S . cerevisiae culture product on the in vitro mixed ruminal microorganism fermentation of ground corn, soluble starch, alfalfa hay, and Coastal bermudagrass hay . In the presence of ground corn, neither concentration (0.35 or 0.73 g/L) of S . cerevisiae culture nor live cells had any effect on final pH, H2, CH4, propionate, or butyrate . The S . cerevisiae culture had no effect on acetate, but both concentrations of S . cerevisiae live cells decreased acetate and the acetate:propionate ratio . When soluble starch was the substrate, both concentrations of S . cerevisiae live cells and 0.73 g/L of S . cerevisiae culture decreased the acetate:propionate ratio . Although the treatment effects were not statistically significant, both concentrations of live cells and 0.73 g/L of the culture decreased lactate concentrations compared with the control incubations . When alfalfa hay served as the substrate, neither the S . cerevisiae culture nor the live cells had an effect on propionate, butyrate, or the acetate:propionate ratio . Both concentrations of S . cerevisiae culture decreased the final pH and in vitro dry matter disappearance, and the 0.73 g/L treatment decreased the amount of acetate . However, both treatments of S . cerevisiae live cells increased final pH and decreased acetate and in vitro dry matter disappearance . Neither yeast treatment had much effect on the Coastal bermudagrass hay fermentations . In general, both S . cerevisiae supplements seemed to have similar effects on the mixed ruminal microorganism fermentation.

J Clin Pediatr Dent, 2002 Fall, 27(1), 1 - 4
Food sugar substitutes: a brief review for dental clinicians; Roberts MW et al.; The frequent ingestion of fermentable sugars such as sucrose, fructose, glucose and maltose is conducive to the development of caries in the teeth of susceptible individuals . Natural and artificial alternatives to these sugars have been and continue to be developed as non/low-caloric sweeteners . The US Food and Drug Administration have approved four non-caloric sweeteners at present . However, there are several other non-caloric sweeteners being commonly used in other countries . A review of these sweeteners is provided with information on a promising new agent that has not yet gained FDA approval.

J Anim Sci, 2002 Oct, 80(10), 2656 - 62
Production of amines in equine cecal contents in an in vitro model of carbohydrate overload; Bailey SR et al.; Acute laminitis can be induced experimentally in horses by the administration of carbohydrate, resulting in fermentation within the cecum and ischemia-reperfusion of the digits . The products of fermentation that trigger acute laminitis are as yet unknown; however, compounds such as amines might play a role due to their potential vasoactive properties . The objectives of this study were to quantify the amines present in equine cecal contents and to use a model of carbohydrate overload in vitro to test the hypothesis that carbohydrate fermentation is associated with increased amine production . Cecal contents from each horse were divided into aliquots and incubated anaerobically with either cornstarch or inulin (a form of fructan carbohydrate; both 1 g/100 mL) . The pH was measured and samples were taken at the same time for amine measurement by HPLC at 2-h intervals over a 24-h period . In a second set of experiments, the effects of the antibiotic virginiamycin (1 mg/100 mL), calcium (CaPO4; 0.3 g/100 mL), and plant steroidal saponin (Yucca schidigera extract; 0.1 g/100 mL) were examined on pH and amine concentrations in cecal contents incubated with starch or inulin . Both starch and inulin caused significant time-dependent falls in pH, from 6.7 +/- 0.1 at 0 h to 5.2 +/- 0.1 (starch) and 5.0 +/- 0.1 (inulin) at 24 h . Fermentation of carbohydrate was also associated with increased production of phenylethylamine and isoamylamine (two- to threefold increases) as well as putrescine and cadaverine (1.5- to twofold increases) . Virginiamycin inhibited the fall in pH and increases in production of phenylethylamine and isoamylamine, while calcium phosphate moderated the changes in pH only . Yucca schidigera extract was without effect . These data show that fermentation of carbohydrate by equine cecal microbiota may lead to increased production of amines.

Biol Trace Elem Res, 2002 Oct, 89(1), 43 - 52
Influence of dietary factors on calcium bioavailability: a brief review; Camara-Martos F et al.; There are several factors that affected calcium bioavailability, such as physiological and dietary factors . These dietary factors help to achieve an appropiate status of calcium for a correct bone mineralization . In this pathway, recently some compounds present in milk that seem improve calcium absorption such as lactose and certain caseinophosphopeptides formed during digestion of caseins have been studied . On the other hand, the possible inhibitatory effect of fiber has been also studied, without conclusive results between in vitro and in vivo studies and the role of phytic acid on impairs calcium bioavailability could be prevented by using fructo-oligosaccharides, which cannot be digested in the small intestine and arrive practically intact to the colon, where are fermented . Finally, calcium fortification must be executed by suitable compounds with high bioavailability, better technological properties, and a correct calcium:phosphorus ratio . For that reason, the objective of the present article is to review the influence of all these conditional factors on calcium bioavailability.

EMBO J, 2002 Nov 1, 21(21), 5653 - 61
Identification and reconstitution of the yeast mitochondrial transporter for thiamine pyrophosphate; Marobbio CM et al.; The genome of Saccharomyces cerevisiae contains 35 members of a family of transport proteins that, with a single exception, are found in the inner membranes of mitochondria . The transport functions of the 15 biochemically identified mitochondrial carriers are concerned with shuttling substrates, biosynthetic intermediates and cofactors across the inner membrane . Here the identification of the mitochondrial carrier for the essential cofactor thiamine pyrophosphate (ThPP) is described . The protein has been overexpressed in bacteria, reconstituted into phospholipid vesicles and identified by its transport properties . In confirmation of its identity, cells lacking the gene for this carrier had reduced levels of ThPP in their mitochondria, and decreased activity of acetolactate synthase, a ThPP-requiring enzyme found in the organellar matrix . They also required thiamine for growth on fermentative carbon sources.

Phytother Res, 2002 Nov, 16(7), 700 - 2
Anti-stress and anti-fatigue effect of fermented rice bran; Kim KM et al.; The anti-stress and anti-fatigue effects of a hot water extract of fermented rice bran (FRB) were investigated with Saccharomyces cerevisae IFO 2346 on rats or mice . Oral administration (1 g/kg/day) of a hot water extract of FRB inhibited major changes in weight of the adrenal, thymus, spleen and thyroid, showing the anti-stress effect . A hot water extract of FRB also inhibited the increase of GPT and LDH activity, cholesterol and serum glucose levels . Administration (1 g/kg/day) for 2 weeks significantly prolonged the swimming time, resulting in an increase of the anti-fatigue effect . From these results, it can be considered that FRB has an anti-stress and anti-fatigue effect .

Dig Liver Dis, 2002 Sep, 34 Suppl 2, S29 - 33
Interactions between novel micro-organisms and intestinal flora; Aureli P et al.; Microbial strains traditionally used to ferment food have a long history of safe use and are, therefore, considered as generally recognised as safe . Many of these micro-organisms have also functional attributes and are included among probiotics . New species and strains of bacteria with desirable technological and functional properties are constantly being identified; in addition, micro-organisms can be engineered by recently developed biotechnological tools in order to accelerate strain improvement . Although the potentialities of novel micro-organisms with better probiotic and technological properties are promising, it cannot be assumed that they share the safety record of traditional micro-organisms, since they may pose unique challenges for human health . The risk assessment and safety evaluation of novel micro-organisms must focus, primarily, on their potential harmful effects, both direct and indirect, upon host resident intestinal microflora . Genetically modified micro-organisms need further assessment for the complete characterisation of the DNA rearrangement and of the final product, in order to establish the "substantial equivalence" with the parental strain.

J Ind Microbiol Biotechnol, 2002 Nov, 29(5), 264 - 7
Bioconversion of acid- and gamma-ray-treated sweet potato residue to microbial protein by mixed cultures; Aziz NH et al.; Sweet potato residue, a starchy agricultural waste, was used as a substrate to produce microbial protein by Fusarium moniliforme and Saccharomyces cerevisiae in submerged fermentation . Acid- and gamma-irradiation-pretreated sweet potato residue enhanced the biomass yield and protein production when the residue was fermented with F . moniliforme and S . cerevisiae . A mixed culture of F . moniliforme and S . cerevisiae efficiently and rapidly utilized free sugars; the maximal biomass yield (13.96 g/l) and protein production (65.8%) were obtained after 3 days fermentation . Lower carbon utilization by the two microbial strains occurred in the waste-containing media as compared to control, increasing the economic value of the waste usage.

J Ind Microbiol Biotechnol, 2002 Nov, 29(5), 221 - 7
Fermentation of sugar mixtures using Escherichia coli catabolite repression mutants engineered for production of L-lactic acid; Dien BS et al.; Conversion of lignocellulose to lactic acid requires strains capable of fermenting sugar mixtures of glucose and xylose . Recombinant Escherichia coli strains were engineered to selectively produce L-lactic acid and then used to ferment sugar mixtures . Three of these strains were catabolite repression mutants (ptsG(-)) that have the ability to simultaneously ferment glucose and xylose . The best results were obtained for ptsG(-) strain FBR19 . FBR19 cultures had a yield of 0.77 (g lactic acid/g added sugar) when used to ferment a 100 g/l total equal mixture of glucose and xylose . The strain also consumed 75% of the xylose . In comparison, the ptsG(+) strains had yields of 0.47-0.48 g/g and consumed 18-22% of the xylose . FBR19 was subsequently used to ferment a variety of glucose (0-40 g/l) and xylose (40 g/l) mixtures . The lactic acid yields ranged from 0.74 to 1.00 g/g . Further experiments were conducted to discover the mechanism leading to the poor yields for ptsG(+) strains . Xylose isomerase (XI) activity, a marker for induction of xylose metabolism, was monitored for FBR19 and a ptsG(+) control during fermentations of a sugar mixture . Crude protein extracts prepared from FBR19 had 10-12 times the specific XI activity of comparable samples from ptsG(+) strains . Therefore, higher expression of xylose metabolic genes in the ptsG(-) strain may be responsible for superior conversion of xylose to product compared to the ptsG(+) fermentations.

Appl Environ Microbiol, 2002 Nov, 68(11), 5750 - 5
Large-scale production of coenzyme F420-5,6 by using Mycobacterium smegmatis; Isabelle D et al.; Production of coenzyme F420 and its biosynthetic precursor FO was examined with a variety of aerobic actinomycetes to identify an improved source for these materials . Based on fermentation costs, safety, and ease of growth, Mycobacterium smegmatis was the best source for F420-5,6 . M . smegmatis produced 1 to 3 micromol of intracellular F420 per liter of culture, which was more than the 0.85 to 1.0 micromol of F420-2 per liter usually obtained with Methanobacterium thermoautotrophicum and approximately 10-fold higher than what was previously reported for the best aerobic actinomycetes . An improved chromatography system using rapidly flowing quaternary aminoethyl ion-exchange material and Florisil was used to more quickly and easily purify F420 than with previous methods.

Appl Environ Microbiol, 2002 Nov, 68(11), 5620 - 4
Expression of an anaplerotic enzyme, pyruvate carboxylase, improves recombinant protein production in Escherichia coli; March JC et al.; Anaplerotic enzyme reactions are those which replenish tricarboxylic acid intermediates that are withdrawn for the synthesis of biomass . In this study, we examined recombinant protein production in Escherichia coli containing activity in an additional anaplerotic enzyme, pyruvate carboxylase . In batch fermentations, the presence of pyruvate carboxylase resulted in 68% greater production of the model protein, beta-galactosidase, 41% greater cell yield, and 57% lower acetate concentration . We discuss why these results indicate that acetate concentration does not limit cell growth and protein synthesis, as predicted by other researchers, and suggest instead that the rate of acetate formation represents an inefficient consumption of glucose carbon, which is reduced by the presence of pyruvate carboxylase.

Appl Environ Microbiol, 2002 Nov, 68(11), 5326 - 35
Molecular analysis of maltotriose transport and utilization by Saccharomyces cerevisiae; Day RE et al.; Efficient fermentation of maltotriose is a desired property of Saccharomyces cerevisiae for brewing . In a standard wort, maltotriose is the second most abundant sugar, and slower uptake leads to residual maltotriose in the finished product . The limiting factor of sugar metabolism is its transport, and there are conflicting reports on whether a specific maltotriose permease exists or whether the mechanisms responsible for maltose uptake also carry out maltotriose transport . In this study, radiolabeled maltotriose was used to show that overexpression of the maltose permease gene, MAL61, in an industrial yeast strain resulted in an increase in the rate of transport of maltotriose as well as maltose . A strain derived from W303-1A and lacking any maltose or maltotriose transporter but carrying a functional maltose transport activator (MAL63) was developed . By complementing this strain with permeases encoded by MAL31, MAL61, and AGT1, it was possible to measure their specific transport kinetics by using maltotriose and maltose . All three permeases were capable of high-affinity transport of maltotriose and of allowing growth of the strain on the sugar . Maltotriose utilization from the permease encoded by AGT1 was regulated by the same genetic mechanisms as those involving the maltose transcriptional activator . Competition studies carried out with two industrial strains, one not containing any homologue of AGT1, showed that maltose uptake and maltotriose uptake were competitive and that maltose was the preferred substrate . These results indicate that the presence of residual maltotriose in beer is not due to a genetic or physiological inability of yeast cells to utilize the sugar but rather to the lower affinity for maltotriose uptake in conjunction with deteriorating conditions present at the later stages of fermentation . Here we identify molecular mechanisms regulating the uptake of maltotriose and determine the role of each of the transporter genes in the cells.

Protein Expr Purif, 2002 Nov, 26(2), 309 - 20
A plasmid system for optimization of Fab' production in Escherichia coli: importance of balance of heavy chain and light chain synthesis; Humphreys DP et al.; We demonstrate the importance of optimizing the balance of light chain (LC) and heavy chain (HC) expression to achieve high level production of Fab' fragments in the Escherichia coli periplasm . The LC:HC balance has been controlled by varying the codon usage of the signal peptide (SP) and 5' mature domain coding regions . Different SP coding regions have been identified from a codon wobble-based library using alkaline phosphatase (AP) as a reporter gene . A plasmid system that enables random combination of these variant SP coding regions is used to construct optimized Fab' expression plasmids . These small plasmid libraries facilitated selection of optimal Fab' expression plasmids and resulted in increases of periplasmic yield, up to 580 mgL(-1) from E . coli fermentations and will enable rapid variable region subcloning and selection of future Fab(') expression plasmids.

Protein Expr Purif, 2002 Nov, 26(2), 249 - 59
Production of salmon calcitonin by direct expression of a glycine-extended precursor in Escherichia coli; Ray MV et al.; The export of heterologous products into the conditioned medium of an Escherichia coli culture offers the advantages of a higher product yield, an increased probability of recovering an intact recombinant protein, proper folding for biological activity, and greater stability of a secreted product . In this report, we describe the development of an optimized direct expression system, designed to maximize the extracellular accumulation of recombinant glycine-extended salmon calcitonin peptide (sCTgly) . We have used dual promoters, an ompA signal sequence, co-expression of homologous secretion factor genes, and multiple gene cartridges to express the sCTgly . High-density fermentation conditions have been developed that allow for the selective secretion and accumulation of the expressed sCTgly at very high levels . Purification and in vitro enzymatic conversion by peptidylglycine alpha-amidating monooxygenase yields authentic, biologically active salmon calcitonin . This recombinant production technology is applicable to a variety of amidated peptide hormones.

J Agric Food Chem, 2002 Nov 6, 50(23), 6798 - 803
Plant-derived biomolecules in fermented cabbage; Tolonen M et al.; The formation of plant-derived biomolecules during sauerkraut fermentation was studied . Cabbage was fermented with a starter culture, and the results were compared to the results of spontaneous fermentation . The concentration of flavonoids and glucosinolates was analyzed by HPLC, and that of the glucosinolate breakdown products, by GC-MS . Of the 20 different flavonoids tested, only kaempferol was found (0.9 mg/ kg FW, fresh weight) . The content of kaempferol remained constant in the cabbage fiber matrix over the fermentation process . The nitrite concentration was below the detection limit in both fermentations . The total glucosinolate content in the raw material was 3.71 micro mol/g DW, dry weight . Glucosinolates were totally decomposed in both fermentations during two weeks, and different types of breakdown products were formed . Isothiocyanates, indole-3-carbinol, goitrin, allyl cyanide, and nitriles were determined in the fermented cabbage . Isothiocyanates and allyl cyanide were the predominant breakdown products in both fermentations . Sulforaphane nitrile and goitrin were found only in small quantities in the end products.

J Microbiol Methods, 2003 Jan, 52(1), 59 - 68
5-Cyano-2,3-ditolyl tetrazolium chloride (CTC) reduction in a mesophilic anaerobic digester: measuring redox behavior, differentiating abiotic reduction, and comparing FISH response as an activity indicator; Gruden CL et al.; The tetrazolium salt 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) has been widely applied to assess microbiological activity in environmental samples . CTC reduction has previously been quantified in a variety of anaerobic systems (i.e., fermentative, nitrate reducing, sulfate reducing) using direct microscopy, solvent extraction, and flow cytometry . In this work, extracellular CTC reduction was observed and distinguished from its intercellular counterparts by the amorphous character and near uniform fluorescence of the resulting formazan precipitates (CTF) . Fluorescence yielded by non-cellular-associated formazan precipitates bleached much more rapidly than CTF formed within cells under identical UV exposure (<2 min) . Dehydrogenase activity assays and fluorescent in situ hybridization (FISH) were simultaneously carried out in microcosms containing active anaerobic digester biomass, propylene glycol, and settled sewage centrate for direct comparison . In substrate limited microcosms, quantitative FISH measurements remained well above their detection limit indicating sustained intercellular ribosomal RNA concentrations over a 5-day period, while dehydrogenase assays (CTC) decreased to background levels within 14 h of substrate limitation . Results from this work suggest that CTC reduction in cell-free samples may impede accurate enzyme activity measurements, particularly when quantification involves solvent extraction, flow cytometry, or software-aided counting . In addition, activity assessment in anaerobic digesters using FISH and CTC reduction assays may be comparable until substrate becomes limited.

Biochem J, 2003 Feb 15, 370(Pt 1), 35 - 46
Proteomic response to physiological fermentation stresses in a wild-type wine strain of Saccharomyces cerevisiae; Trabalzini L et al.; We report a study on the adaptive response of a wild-type wine Saccharomyces cerevisiae strain, isolated from natural spontaneous grape must, to mild and progressive physiological stresses due to fermentation . We observed by two-dimensional electrophoresis how the yeast proteome changes during glucose exhaustion, before the cell enters its complete stationary phase . On the basis of their identification, the proteins representing the S . cerevisiae proteomic response to fermentation stresses were divided into three classes: repressed proteins, induced proteins and autoproteolysed proteins . In an overall view, the proteome adaptation of S . cerevisiae at the time of glucose exhaustion seems to be directed mainly against the effects of ethanol, causing both hyperosmolarity and oxidative responses . Stress-induced autoproteolysis is directed mainly towards specific isoforms of glycolytic enzymes . Through the use of a wild-type S . cerevisiae strain and PMSF, a specific inhibitor of vacuolar proteinase B, we could also distinguish the specific contributions of the vacuole and the proteasome to the autoproteolytic process.

FEMS Microbiol Lett, 2002 Oct 8, 215(2), 169 - 74
High expression of a sucrose non-fermenting (SNF1)-related protein kinase from Colletotrichum gloeosporoides f . sp . malvae is associated with penetration of Malva pusilla; Goodwin PH et al.; A sucrose non-fermenting (SNF1)-related protein kinase homologue, cgsnf, from Colletotrichum gloeosporoides f . sp . malvae, a hemibiotrophic fungal pathogen of round-leaved mallow (Malva pusilla) was examined . During infection, cgsnf showed a large peak in expression relative to a constitutively expressed fungal actin gene when appressoria had formed during the penetration phase and then showed much lower expression levels during subsequent necrotrophic growth in the host . In pure culture with glucose or glycerol as sole carbon sources, expression levels were similar to that during necrotrophic growth . Expression was consistently higher in glycerol than in glucose cultures, which may reflect a lower cellular energy status in the fungus . These results are consistent with cgsnf having a role in transmitting nutritional signals, which may be involved with host penetration.

J Nat Prod, 2002 Oct, 65(10), 1491 - 3
Panepophenanthrin, from a mushroom strain, a novel inhibitor of the ubiquitin-activating enzyme; Sekizawa R et al.; Screening for inhibitors of the ubiquitin-proteasome pathway, considered to regulate important cellular events and linked to serious diseases as well, led to isolation of a new compound, panepophenanthrin, from the fermented broth of a mushroom strain, Panus rudis Fr . IFO 8994 . This is the first inhibitor of the ubiquitin-activating enzyme, which is indispensable for the ubiquitin-proteasome pathway . The structure of panepophenanthrin was determined by NMR and X-ray crystallographic analyses as 1,3a,10-trihydroxy-10c-(3-hydroxy-3-methylbut-1-enyl)-5,5-dimethyl-1,2,3,3a,5,5a,8,9,10,10a,10b,10c-dodecahydro-4-oxa-2,3,8,9-diepoxyacephenanthrylen-7-one.

Int J Med Microbiol, 2002 Sep, 292(3-4), 207 - 14
Identification and distribution of the enterohemorrhagic Escherichia coli factor for adherence (efa1) gene in sorbitol-fermenting Escherichia coli O157: H-; Janka A et al.; Sorbitol-fermenting (SF) Shiga toxin (Stx)-producing Escherichia coli (STEC) O157:H- strains are emerging as causes of hemorrhagic colitis and the hemolytic-uremic syndrome in Europe . Using subtractive hybridization between SF STEC O157:H- strain 493/89 and STEC O157:H7 strain EDL933, three different fragments, of approximately 700 bp in length, were identified . Each demonstrated > 99% homology to genes encoding the enterohemorrhagic E . coli factor for adherence (efa1) and lymphostatin (lifA) . Therefore, a cosmid library was constructed from SF STEC O157:H- strain 493/89, and one clone containing these fragments was sequenced . This sequencing demonstrated a 9669-bp open reading frame (ORF) that had 99.9% sequence homology to efa1 of STEC O111:H- strain E45035 and to lifA of an enteropathogenic E . coli O127:H6 strain E2348/69 . In STEC O157:H7 strain EDL933, only small (ca . 3 kb) initial and terminal fragments of this ORF are present . PCR analysis with primers complementary to the efa1/lifA sequence of strain 493/89 indicated that the complete sequence is present in each of 10 SF STEC O157:H- isolates but in none of 10 STEC O157:H7 strains investigated . The presence of the complete efa1/lifA also in both tested E . coli O55:H7 strains supports the hypothesis that SF STEC O157:H- are phylogenetically closer to the proposed E . coli O55:H7 ancestor than STEC O157:H7 . Our data demonstrate the presence of a potential virulence gene in SF STEC O157:H- that is only rudimentarily present in STEC O157:H7.

FASEB J, 2002 Dec, 16(14), 1958 - 60 Epub 2002 Oct 18.
Inhibition of the PDGF receptor by red wine flavonoids provides a molecular explanation for the "French paradox"; Rosenkranz S et al.; The mortality rate from coronary artery disease (CAD) in France is approximately 50% compared to other European countries and the United States ("French paradox") . Epidemiological studies indicate an inverse relationship between moderate wine consumption and CAD mortality . Here, we demonstrate that preincubation of vascular smooth muscle cells (VSMCs) with red wine, but not white wine, inhibits ligand binding and the subsequent tyrosine phosphorylation of the platelet-derived growth factor beta receptor (betaPDGFR), which plays a critical role in the pathogenesis of atherosclerosis . As a consequence, red wine abrogates the ligand-induced recruitment of betaPDGFR-associated signaling molecules (RasGAP, SHP-2, PI3K, PLCgamma), PDGF-dependent downstream events such as Erk activation and induction of immediate early genes, and VSMC proliferation and migration . Wine analysis revealed flavonoids of the catechin family as major constituents of red wine, and these were identified as potent inhibitors of betaPDGFR signaling . Importantly, the concentrations of red wine/catechins shown to inhibit the PDGFR in vitro correlate with the serum levels after red wine consumption in humans . We conclude that nonalcoholic constituents of red wine, which accumulate during the "mash fermentation," inhibit betaPDGFR activation and PDGF-dependent cellular responses in VSMCs . Therefore, catechin-mediated inhibition of betaPDGFR signaling offers a molecular explanation for the "French paradox."

Food Addit Contam, 2002 Sep, 19(9), 810 - 8
Determination of ivermectin and moxidecin residues in bovine milk and examination of the effects of these residues on acid fermentation of milk; Imperiale F et al.; Ivermectin (IVM) and moxidectin (MXD) are broad-spectrum antiparasitic drugs not approved for use in dairy animals, although their use in dairy sheep, goats and cattle nevertheless occurs in many parts of the world . The work reported here describes (1) the application of an HPLC method (including milk samples clean-up and chemical extraction) to quantify IVM and MXD residues in bovine milk, and (2) an assessment of the effect of different IVM and MXD concentrations on bovine milk acid fermentation . The latter was carried out using the 'yoghurt test' to determine the minimum IVM and MXD concentrations affecting milk acid fermentation . The sample clean-up, chemical extraction and the validated HPLC method allowed the quantification of IVM and MXD up to 0.1 ng ml(-1) in milk with acceptable validation coefficients . Drug recoveries from fortified milk samples ranged between 72% (CV = 9.1%) and 75% (CV = 13.3%) for MXD and IVM, respectively . Neither IVM nor MXD affected the acid fermentation of bovine milk . In fact, there was no drug-induced changes on milk acidity even at IVM and MXD concentrations as high as 1000 ng ml(-1) . These results indicate that the yoghurt biological test is not suitable to evaluate the presence of milk residues for these antiparasitic compounds . Thus, a highly sensitive HPLC technique is the only reliable method for determining the presence of residual concentrations of IVM and MXD in milk and dairy products to assure consumer safety.

J Microencapsul, 2002 Jul-Aug, 19(4), 397 - 405
Cultivation and modelling of encapsulated Saccharomyces cerevisiae in NaCS-PDMDAAC polyelectrolyte complexes; Mei LH et al.; The cultivation of encapsulated S . cerevisiae in NaCS-PDMDAAC polyelectrolyte complexes was studied . The results showed that the encapsulated microorganisms had the same growth trends as in its free cell culture and, thus, NaCS-PDMDAAC microcapsules were suitable for the encapsulation of these biological substances . The encapsulated S . cerevisiae cells were fermented sequentially for 16 batches . The highest cell density in the capsules reached 2.64 x 10(10) cells mL(-1) and the ethanol concentration was 47.0 g L(-1) . A model of the cultivation of the encapsulated S . cerevisiae was developed.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 453 - 61
Production of alpha-amylase with Aspergillus oryzae on spent brewing grain by solid substrate fermentation; Bogar B et al.; Ten Aspergillus oryzae strains were screened in solid substrate fermentation for alpha-amylase production on spent brewing grain (SBG) and on corn fiber . SBG proved to be a better substrate for enzyme production than corn fiber . A Plackett-Burman experimental design was used to optimize the medium composition for the best strain . Solid substrate fermentation on optimized medium with A . oryzae NRRL 1808 (=ATCC 12892) strain in stationary 500-mL Erlenmeyer flask culture yielded 4519 U of alpha-amylase/g of dry matter substrate in 3 d . The whole solid substrate fermentation material (crude enzyme, in situ enzyme) may be considered a cheap biocatalytic material for animal feed rations and for bioalcohol production from starchy materials.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 415 - 29
Mn2+ alters peroxidase profiles and lignin degradation by the white-rot fungus Pleurotus ostreatus under different nutritional and growth conditions; Cohen R et al.; The white-rot fungus Pleurotus ostreatus produces two types of extracellular peroxidases: manganese-dependent peroxidase (MnP) and versatile peroxidase (VP) . The effect of Mn2+ on fungal growth, peroxidase activity profiles, and lignin degradation by P . ostreatus was studied in liquid culture and under solid-state fermentation conditions on perlite, the latter resembling the natural growth conditions of this fungus . The fungus was grown in either a defined asparagine-containing basidiomycete selective medium (BSM) or in a rich peptone medium (PM) . Biomass production, as determined by respiration experiments in solid-state fermentation and liquid cultures and fungal growth on Petri dishes, was higher in the PM than in the BSM . Mn2+ affected biomass production only in the PM on Petri dishes . In the nonamended PM, high levels of MnP and VP activity were detected relative to the nonamended BSM . Nevertheless, a higher rate of 14C-lignin mineralization was measured in the Mn2+-amended BSM, as determined during the course of 47 d of fermentation . Mn2+ amendment of the PM increased mineralization rate to that obtained in the Mn2+-amended BSM . The enzyme activity profiles of MnP and VP were studied in the BSM using anion-exchange chromatography . In the nonamended BSM, only minute levels of MnP and VP were detected . On Mn2+ amendment, two MnP isoenzymes (B1 and B2) appeared . Isoenzyme B2 was purified and showed 100% identity with the MnP isoenzyme purified in our previous study from PM-solid-state fermentation (P6) . P6 was found to be the dominant isoenzyme in terms of activity level and gene expression compared with the VP isoenzymes . Based on these results, we concluded that Mn2+ plays a key role in lignin degradation under different nutritional and growth conditions, since it is required for the production of MnP in P . ostreatus.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 251 - 60
Solid-state fermentation for production of phytase by Rhizopus oligosporus; Sabu A et al.; Solid-state fermentation of coconut oil cake has been carried out with Rhizopus oligosporus for the production of phytase . Phytase is used commercially in the animal feed industry to improve animal performance because there is a substantial and growing interest among swine and poultry producers in the application of phytase to improve the nutritional quality in animal feeds . Demonstrated benefits include improved feed yield ratios and reduction in the environmental costs associated with the disposal of animal wastes . We report the production of extracellular phytase by R . oligosporus under solid-state fermentation using coconut oil cake as substrate . Maximal enzyme production (14.29 U/g of dry substrate) occurred at pH 5.3, 30 degrees C, and 54.5% moisture content after 96 h of incubation . The addition of extra nutrients to the substrate resulted in inhibition of product formation . The results indicate the scope for production of phytase using coconut oil cake as solid substrate without additional nutrients.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 193 - 9
Effect of pH on simultaneous saccharification and isomerization by glucoamylase and glucose isomerase; Mishra A et al.; pH and temperature play critical roles in multistep enzymatic conversions . In such conversions, the optimal pH for individual steps differs greatly . In this article, we describe the production of glucoamylase (from Aspergillus oryzae MTCC152 in solid-state fermentation) and glucose isomerase (from Streptomyces griseus NCIM2020 in submerged fermentation), used in industries for producing high-fructose syrup . Optimum pH for glucoamylase was found to be 5.0 . For glucose isomerase, the optimum pH ranged between 7.0 and 8.5, depending on the type of buffer used . Optimum temperature for glucoamylase and glucose isomerase was 50 and 60 degrees C, respectively . When both the enzymatic conversions were performed simultaneously at a compromised pH of 6.5, both the enzymes showed lowered activity . We also studied the kinetics at different pHs, which allows the two-step reaction to take place simultaneously . This was done by separating two steps by a thin layer of urease . Ammonia generated by the hydrolysis of urea consumed the hydrogen ions, thereby allowing optimal activity of glucose isomerase at an acidic pH of 5.0.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 179 - 91
Gibberellic acid production by solid-state fermentation in coffee husk; Machado CM et al.; Five strains of Gibberella fujikuroi and one of Fusarium moniliforme were screened for the production of gibberellic acid (GA3) in coffee husk, and based on the results, one strain, G . fujikuroi LPB-06, was selected . The comparative production of GA3 by solid-state fermentation and submerged fermentation indicated better productivity with the former technique, mainly with pretreated substrate . The GA3 accumulation was 6.1 times higher in the case of solid-state fermentation . Considering the C:N relation, higher yields of GA3 were achieved using a mixed substrate comprising coffee husk and cassava bagasse (7:3, dry wt), increasing the results twice . Supplementation of an optimized saline solution containing 0.03% FeSO4 and 0.01% (NH4)2SO4 enhanced the accumulation of GA3 1.7 times in the fermented substrate . Under the finally optimized condition, the culture gave a maximum of 492.5 mg of GA3/kg of dry substrate, with a pH of 5.3, moisture of 75%, and incubation temperature of 29 degrees C . GA3 yield was almost 13 times more than the initial results.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 169 - 77
Relationship between coffee husk caffeine degradation and respiration of Aspergillus sp . LPBx in solid-state fermentation; Brand D et al.; Studies were carried out in a packed-bed column fermentor using coffee husk as substrate in order to verify a relationship between caffeine degradation and the respiration of Aspergillus sp . LPBx . Fermentation conditions were optimized by using factorial design experiments . The kinetic study showed that the caffeine degradation was related to the development of mold and its respiration and also with the consumption of reducing sugars present in coffee husk . From the values obtained experimentally for oxygen uptake rate and CO2 evolved, we determined a biomass yield of 3.811 g of biomass/g of consumed O2 and a maintenance coefficient of 0.0031 g of consumed O2/(g of biomass x h) . The maximum caffeine degradation achieved was 90%.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 129 - 40
Effect of environmental factors and carbohydrate on gellan gum production; Kanari B et al.; Submerged culture fermentation studies were carried out in batch mode for optimizing the environmental parameters and carbon source requirement by Pseudomonas elodea for the production of gellan gum . The maximum production of gellan gum was obtained with 16-h-old culture and 8% inoculum at 30 degrees C and pH 7.0 after 52 h of incubation (6.0 g/L) . Of the various carbon sources tested, 2% sucrose, glucose, and soluble starch yielded considerably high amounts of gellan . Studies on the concentration of various carbohydrates on gellan gum production indicated that the optimum concentration of glucose and starch was 3%, whereas for sucrose it was 4% . The addition of glucose in the medium above 3% had a detrimental effect on gellan yield . The investigation of intermediate two-step addition of glucose under identical conditions of fermentation showed an enhanced production of gellan (8.12 g/L) as compared with the control (6.0 g/L) . To optimize the recovery of gellan from fermented broth, different solvents were tested for precipitation of gellan gum . Among the various solvents tested, tetrahydrofuran gave better recovery of gellan (82%) as compared with the conventional solvent isopropanol (49%).

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 119 - 28
Optimization of media by evolutionary algorithms for production of polyols; Patil SV et al.; Biotransformation of sucrose-based medium to polyols has been reported for the first time using osmophilic yeast, Hansenula anomala . A new, real coded evolutionary algorithm was developed for optimization of fermentation medium in parallel shake-flask experiments . By iteratively employing the nature-inspired techniques of selection, crossover, and mutation for a fixed number of generations, the algorithm obtains the optimal values of important process variables, namely, inoculum size and sugar, yeast extract, urea, and MgSO4 concentrations . Maximum polyols yield of 76.43% has been achieved . The method is useful for reducing the overall development time to obtain an efficient fermentation process.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 109 - 18
Production of ligninolytic enzymes for dye decolorization by cocultivation of white-rot fungi Pleurotus ostreatus and phanerochaete chrysosporium under solid-state fermentation; Verma P et al.; Lignocellulosic wastes such as neem hull, wheat bran, and sugarcane bagasse, available in abundance, are excellent substrates for the production of ligninolytic enzymes under solid-state fermentation by white-rot fungi . A ligninolytic enzyme system with high activity showing enhanced decomposition was obtained by cocultivation of Pleurotus ostreatus and Phanerochaete chrysosporium on combinations of lignocellulosic waste . Among the various substrate combinations examined, neem hull and wheat bran wastes gave the highest ligninolytic activity . A maximum production of laccase of 772 U/g and manganese peroxidase of 982 U/g was obtained on d 20 and lignin peroxidase of 656 U/g on d 25 at 28 +/- 1 degrees C under solid-state fermentation . All three enzymes thus obtained were partially purified by acetone fractionation and were exploited for decolorizing different types of acid and reactive dyes.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 63 - 70
Invertase production on solid-state fermentation by Aspergillus niger strains improved by parasexual recombination; Montiel-Gonzalez AM et al.; Invertase production by Aspergillus niger grown by solid-state fermentation was found to be higher than by conventional submerged fermentation . The haploid mutant strains Aw96-3 and Aw96-4 showed better productivity of various enzymes, as compared to wild-type parental strain A . niger C28B25 . Here we use parasexual crosses of those mutants to increase further the productivity of invertase in solid-state fermentation . We isolated both a diploid (DAR2) and an autodiploid (AD96-4) strain, which were able to grow in minimal medium after mutation complementation of previously isolated haploid auxotrophic strains . Invertase production was measured in solid-state fermentation cultures, using polyurethane foam as an inert support for fungal growth . Water activity value (Aw) was adjusted to 0.96, since low Aw values are characteristic in some solid-state fermentation processes . Such diploid strains showed invertase productivity levels 5-18 times higher than levels achieved by the corresponding haploid strains . For instance, values for C28B25, Aw96-3, Aw96-4, DAR2, and AD96-4 were 441, 254, 62, 1324, and 2677 IU/(L x h), respectively . These results showed that genetic recombination, achieved through parasexual crosses in A . niger, results in improved strains with potential applications for solid-state fermentation processes.

Appl Biochem Biotechnol, 2002 Jul-Dec, 102-103(1-6), 49 - 62
Fermentation diagnosis by multivariate statistical analysis; Bicciato S et al.; During the course of fermentation, online measuring procedures able to estimate the performance of the current operation are highly desired . Unfortunately, the poor mechanistic understanding of most biologic systems hampers attempts at direct online evaluation of the bioprocess, which is further complicated by the lack of appropriate online sensors and the long lag time associated with offline assays . Quite often available data lack sufficient detail to be directly used, and after a cursory evaluation are stored away . However, these historic databases of process measurements may still retain some useful information . A multivariate statistical procedure has been applied for analyzing the measurement profiles acquired during the monitoring of several fed-batch fermentations for the production of erythromycin . Multivariate principal component analysis has been used to extract information from the multivariate historic database by projecting the process variables onto a low-dimensional space defined by the principal components . Thus, each fermentation is identified by a temporal profile in the principal component plane . The projections represent monitoring charts, consistent with the concept of statistical process control, which are useful for tracking the progress of each fermentation batch and identifying anomalous behaviors (process diagnosis and fault detection).

J Pediatr Gastroenterol Nutr, 2002 Oct, 35(4), 545 - 50
Variable effects of short chain fatty acids and lactic acid in inducing intestinal mucosal injury in newborn rats; Lin J et al.; BACKGROUND: Short chain fatty acids and lactic acid are colonic bacterial fermentation products . METHODS: To evaluate the effects of these organic acids on the intestinal mucosa, a total of 72 newborn Sprague-Dawley rats (10 days old) were studied . A 3.5F catheter was inserted per rectum 4.0 cm deep into the proximal colon for organic acid administration at a volume of 0.1 ml/10 g body weight . The pH of organic acid solutions and normal saline was adjusted to 4.0 . Group 1 (n = 10) received normal saline as a control . Group 2 (n = 11) received 150 mM acetic acid . Group 3 (n = 11) received 300 mM acetic acid . Group 4 (n = 10) received 150 mM butyric acid . Group 5 (n = 11) received 300 mM butyric acid . Group 6 (n = 7) received 150 mM lactic acid, and group 7 (n = 12) received 300 mM lactic acid . Animals were killed 24 hours after colonic installation of test solutions . RESULTS: Both 300 mM acetic acid and 300 mM butyric acid were associated with impaired weight gain, increased colon wet weight, and increased histologic injury scores in the colon and distal ileum (P < 0.05, analysis of variance) . Both 150 mM acetic acid and butyric acid at 150 mmol/L induced minimal injury in the colon and distal ileum . Neither 150 mM nor 300 mM lactic acid induced any identifiable gross or microscopic intestinal mucosal injury . CONCLUSION: Luminal short chain fatty acids can induce dose-dependent intestinal mucosal injury in newborn rats, resembling the pathology seen in neonatal necrotizing enterocolitis . Overproduction/accumulation of short chain fatty acids, but not lactic acid, in the proximal colon and/or distal ileum may play a role in the pathogenesis of necrotizing enterocolitis in premature infants.

Eur J Cancer Prev, 2002 Oct, 11(5), 457 - 63
Dairy foods and risk of breast cancer: a case-control study in Montevideo, Uruguay; Ronco AL et al.; In order to investigate possible associations of milk and dairy products and the risk of breast cancer (BC) in Montevideo, the authors performed a case-control study in the period 1999-2001 . A total of 333 women were interviewed with a specific questionnaire; 111 of them had been diagnosed with BC and 222 were frequency-matched healthy women, with a recent normal mammography (BIRADS 1) . The questionnaire included a detailed 120-item food-frequency section, as well as questions related to socio-demographic, reproductive, familial, medical and lifestyle variables . There was particular emphasis on types of milk and dairy products . After controlling for age, years of urban status, education, body mass index, age at menarche, menopausal status, family history of BC, number of childbirths, total energy and total fruits, a multivariate analysis found that high intakes of whole milk, chocolate milk and Gruyere cheese were associated with significant increased risk of BC, whereas ricotta cheese and skim yoghurt were associated with significant decreased risks . Low-fat and fermented products combined appear to be the most protective dairy foods . The results suggest that separate analyses for types of milk and cheese, as well as for dairy products in general should be performed in the future.

J Appl Microbiol, 2002, 93(5), 877 - 83
Effects of low electric treatment on yeast microflora; Ranalli G et al.; AIMS: To contribute to the understanding of phenomena related to different intensity electric current treatments on the growth and metabolism of selected micro-organisms using laboratory samples of pure and co-cultures (Saccharomyces cerevisiae strain 404 and Hanseniaspora guilliermondii strain 465) . METHODS AND RESULTS: Low electric current (10, 30, 50 and 100 mA) was applied to prepared samples . Parameters, such as polarity, treatment duration (18-48 h) and type of inoculum yeast, were varied one at a time to highlight their cause-effect relationships . The effects on cell activity as well as microflora viability were assessed . Bioindicators capable of describing the phenomena caused by the electric current on the microflora were identified . CONCLUSIONS: Results demonstrated that a low voltage treatment using graphite electrodes had a greater effect on the viable S . cerevisiae strain 404 microflora . There was less bactericidal activity in the S . cerevisiae strain 404 than in the H . guilliermondii strain 465 . SIGNIFICANCE AND IMPACT OF THE STUDY: These results may be of significant importance in the development of new technological processes in the fields of agriculture and food, particularly new fermenting process controls.

Arch Tierernahr, 2002 Apr, 56(2), 67 - 91
Consequences of the ban of by-products from terrestrial animals in livestock feeding in Germany and the European Union: alternatives, nutrient and energy cycles, plant production, and economic aspects; Rodehutscord M et al.; Consequences of the ban of meat and bone meal (MBM) and animal fat with regard to livestock feeding, cropping, ecology and economy where investigated with an inter-disciplinary approach for Germany and the European Union . Calculations were made for different production systems with pigs and poultry on the basis of statistical data for the production and for the feed markets as well as from requirement data for the respective species and production system . (1.) The ban of MBM from feeding caused a need for alternative protein sources . If all the amount of protein from MBM is to be replaced by soybean meal, in Germany and the EU about 0.30 and 2.30 x 10(6) t would be needed each year (supplementary amino acids not considered) . Alternatively, doubling the grain legume acreage in Germany to about 420,000 ha would supply a similar amount of protein . A wider application of phase feeding with adjusted dietary amino acid concentrations, however, would allow for saving protein to an extent which is similar to the amount of protein that was contributed by MBM in recent years . Thus, the ban is a minor problem in terms of ensuring amino acid supply . (2.) However, alternative plant ingredients cannot compensate for the gap in P supply that is caused by the ban . An additional demand for inorganic feed phosphates of about 14,000 and 110,000 t per year is given in Germany and the EU, respectively . So far, this gap is filled almost completely by increased mining of rock phosphates . Alternatively, a general application of microbial phytase to all diets would largely fill this gap . Until the ban, MBM contributed to 57% of the supplementation of P that was needed for pigs and poultry . The ban of MBM makes large amounts of P irreversibly disappearing from the food chain . (3.) Energy from slaughter offal and cadavers can be utilized in different technologies, in the course of which the efficiency of energy utilisation depends on the technology applied . It is efficient in the cement work or rotation furnace if heat is the main energy required . In contrast, the energetic efficiency of fermentation is low . (4.) Incineration or co-incineration of MBM and other by-products causes pollution gas emissions amounting to about 1.4 kg CO2 and 0.2 kg NOx per kg . The CO2 production as such is hardly disadvantageous, because heat and electrical energy can be generated by the combustion process . The prevention of dangerous gaseous emissions from MBM burning is current standard in the incineration plants in Germany and does not affect the environment inadmissibly . (5.) The effects of the MBM ban on the price for compound feed is not very significant . Obviously, substitution possibilities between different feed ingredients helped to exchange MBM without large price distortions . However, with each kg MBM not used in pig and poultry feeding economic losses of about 0.14 {symbol: see text} have to considered . In conclusion, the by far highest proportion of raw materials for MBM comes as by-products from the slaughter process . Coming this way, and assuring that further treatment is safe from the hygienic point of view, MBM and animal fat can be regarded as valuable sources of amino acids, minerals and energy in feeding pigs and poultry . Using them as feedstuffs could considerably contribute to the goal of keeping limited nutrients, phosphorus in particular, within the nutrient cycle and dealing responsible with limited resources.

Curr Opin Crit Care, 2002 Apr, 8(2), 145 - 51
Gut microbial ecology in critical illness: is there a role for prebiotics, probiotics, and synbiotics?
Bengmark S.
Approximately 70% of the immune system is localized in the gastrointestinal tract: its glands, mucosa, and mucosa-associated lymphoid system . The system influences health conditions because it produces large amounts of important gastrointestinal secretions as rich as breast milk in health-supporting and disease-preventing factors, and because of its rich gastrointestinal flora . The intestine normally contains 10 times more microbes than there are eukaryotic cells in the entire body . The optimal function of these microbes depends on the supply of food destined for the colonic bacteria (fermentable fibers, complex proteins, gastrointestinal secretions) . The consideration of these functions influences outcome . Unfortunately, the conditions (supply of drugs-especially antibiotics, and reduced supply of food-especially fruits and vegetables) in the modern ICU are extremely poor both for optimal gastrointestinal secretion and for flora and need more attention . To improve treatment, a supply of new and effective flora (probiotics) and food for the flora (prebiotics) is needed, from which numerous health-supporting products (synbiotics) will be produced and absorbed at the level of the mucosa, mainly in the lower gastrointestinal tract.

Int J Antimicrob Agents, 2002 Oct, 20(4), 293 - 6
In vitro antimycoplasmal activity of oleuropein; Furneri PM et al.; The activity of oleuropein, a phenolic glycoside contained in olive oil, was investigated in vitro against Mycoplasma hominis, Mycoplasma fermentans, Mycoplasma pneumoniae and Mycoplasma pirum . Oleuropein inhibited mycoplasmas at concentrations from 20 to 320 mg/l . The MICs of oleuropein to M . pneumoniae, M . pirum, M . hominis and M . fermentans were 160, 320, 20 and 20 mg/l, respectively.

Naturwissenschaften, 2002 Oct, 89(10), 433 - 44 Epub 2002 Sep 11.
Biosensors: new approaches in drug discovery; Keusgen M; The development of biosensors for analytical purposes has attracted a great deal of attention in recent years . A biosensor is defined as an analytical device consisting of a biological component (e.g., enzyme, antibody, entire cell, DNA) and a physical transducer (e.g., electrode, optical device) . Biosensors are mostly designed for routine analysis, such as clinical diagnosis, quality control of food, in-process control of fermentations, and in environmental analysis . Many of these sensors are also suitable for screening purposes in order to find new drugs . Such systems should yield information either about compounds with known bioactivity or about the bioactivity of samples with known or unknown chemical composition . Biosensors intended for the latter purpose are essentially based on whole cells carrying receptors and ion channels at their surfaces . Miniaturization of structures, primarily based on silicon, allows integration of many sensors into arrays, which may be suitable for the screening of natural and chemical products as well as combinatorial libraries . Until now, no commercially available sensors of this kind exist but they are expected in the near future . Different biosensors, based on enzymes, antibodies, cells, artificial membranes and entire animal tissues, which can be used in drug discovery and may lead to efficient screening systems in the future, are described in this review.

J Clin Neurosci, 2002 Sep, 9(5), 525 - 9
High frequency of systemic mycoplasmal infections in Gulf War veterans and civilians with Amyotrophic Lateral Sclerosis (ALS); Nicolson GL et al.; The presence of systemic mycoplasmal infections in the blood of Gulf War veterans (n=8) and civilians (n=28) with Amyotrophic Lateral Sclerosis (ALS) and age matched controls (n=70) was investigated by detecting mycoplasma gene sequences with forensic Polymerase Chain Reaction (PCR) and back hybridization with a radiolabeled internal oligonucleotide probe . Almost all ALS patients (30/36 or approximately 83%) showed evidence of Mycoplasma species in blood samples, whereas <9% of controls had blood mycoplasmal infections (P<0.001) . Using PCR ALS patients with a positive test for any mycoplasmal infection were investigated for the presence of M . fermentans, M . pneumoniae, M . hominis and M . penetrans in their blood . All Gulf War veterans with ALS were positive for M . fermentans, except one that was positive for M . genitalium . In contrast, the 22/28 civilians with detectable mycoplasmal infections had M . fermentans (13/22, 59%) as well as other Mycoplasama species in their blood, and two of the civilian ALS patients had multiple mycoplasma species (M . fermentans plus M . hominis) . Of the few control patients that were positive, only two patients (2/70, 2.8%) were positive for M . fermentans (P<0.001) . The results support the suggestion that infectious agents may play a role in the pathogenesis and/or progression of ALS, or alternatively ALS patients are extremely susceptible to systemic mycoplasmal infections.

Int J Food Microbiol, 2002 Nov 15, 79(1-2), 75 - 83
Fermentation and pathogen control: a risk assessment approach; Adams M et al.; Food fermentation has a long tradition of improving the safety, shelf life and acceptability of foods . Although fermented foods generally enjoy a well-founded reputation for safety, some notable outbreaks of foodborne illness associated with fermented foods have occurred . Microbiological risk assessment (MRA), as it has emerged in recent years, provides the scientific basis for the control and management of risk . Aspects of fermented food processes are discussed under the various stages of risk assessment and data are presented that would inform more detailed risk assessments.

Semin Hematol, 2002 Oct, 39(4), 263 - 9
African iron overload; Gordeuk VR; Iron overload is common in rural sub-Saharan African populations that have the custom of drinking a traditional fermented beverage with high iron content . As with both excessive alcohol exposure and HFE hemochromatosis, hepatic portal fibrosis and micronodular cirrhosis are prominent sequelae of African iron overload . Two observations are therefore important in characterizing iron overload in Africa . First, the hepatic iron concentrations associated with African iron overload often far exceed those seen in alcoholic liver disease and histologic changes of alcohol effect are almost always absent . Second, the pattern of iron accumulation in African dietary iron is prominent in both macrophages and hepatic parenchymal cells; this pattern is in contrast to HFE homochromatosis, which is marked by predominantly parenchymal iron-loading . For a long time, it was thought that African iron overload was purely dietary in nature, that increased iron and alcohol in the diet could fully explain markedly elevated tissue iron levels sometimes seen with this condition . Recent studies of pedigrees suggest that, in addition to high dietary iron content, a genetic defect may also be implicated in iron overload in Africans . These studies indicate that the possible defect is different from mutations in the HFE gene frequently found in Caucasians with iron overload, but the putative gene has not been identified . Recent studies also indicate that non-HFE iron overload occurs in African-Americans, but the prevalence and possible genetic basis is yet to be determined .

Appl Microbiol Biotechnol, 2002 Oct, 60(1-2), 186 - 91 Epub 2002 Sep 06.
Kinetic studies on glucose and xylose transport in Saccharomyces cerevisiae; Lee WJ et al.; Zero trans-influx assays of glucose and xylose were performed using Saccharomyces cerevisiae to investigate transport characteristics under high and low glucose conditions . Under high glucose conditions, most glucose was transported by the low-affinity transporter . The high-affinity transporter was expressed under low glucose conditions, transporting over 50% glucose . Inhibition kinetics revealed that xylose was transported by both high- and low-affinity glucose transporters . Affinities of both glucose transporters for xylose were very low under high glucose condition but increased to a similar level to glucose under low glucose condition . The maximum rate of xylose transport increased by 85%, while an overall maximum glucose transport rate decreased by 42% under low glucose condition, indicating the presence of other transport system for sugars except for glucose . It was suggested that expression of the high-affinity transporter and increased affinity of glucose transporters for xylose under low glucose condition would provide a fermentation strategy for enhancing the productivity of xylitol by recombinant S . cerevisiae harboring the xylose reductase gene.

Appl Microbiol Biotechnol, 2002 Oct, 60(1-2), 154 - 9 Epub 2002 Aug 22.
Poly(ethylene glycol)-mediated molar mass control of short-chain- and medium-chain-length poly(hydroxyalkanoates) from Pseudomonas oleovorans; Ashby RD et al.; Three strains of Pseudomonas oleovorans, a well known poly(hydroxyalkanoate) (PHA) producer, were tested for the ability to control PHA molar mass and end group structure by addition of poly(ethylene glycol) (PEG) to the fermentation medium . Each strain of P . oleovorans - NRRL B-14682 (B-14682), NRRL B-14683 (B-14683), and NRRL B-778 (B-778) - synthesized a different type of PHA from oleic acid when cultured under identical growth conditions . Strain B-14682 produced poly(3-hydroxybutyrate) (PHB), while B-14683 synthesized a medium-chain-length PHA ( mcl-PHA) with a repeat unit composition ranging from C4 to C14 and some mono-unsaturation in the C14 alkyl side chains . Strain B-778 synthesized a mixture of PHB (95 mol%) and mcl-PHA (5 mol%) . The addition of 0.5% (v/v) PEG (M(n) =200 g/mol, PEG-200) to the fermentation broth of strains B-14682 and B-778 resulted in chain termination through esterification at the carboxyl terminus of the PHB with PEG chain segments, thus reducing the molar mass by 54% and 23%, respectively . The molar mass of the mcl-PHA produced by strains B-14683 and B-778 also showed a 34% and 47% reduction in the presence of PEG-200, respectively, but no evidence of esterification was present . PEG-400 (M(n) =400 g/mol) had a reduced effect on PHA molar mass . In fact, the molar masses of the mcl-PHA derived from strain B-14683 and both the PHB and mcl-PHA from B-778 were unchanged by PEG-400 . In contrast, the PHB produced by B-14682 showed a 35% reduction in molar mass in the presence of PEG-400.

Appl Microbiol Biotechnol, 2002 Oct, 60(1-2), 139 - 46 Epub 2002 Aug 22.
MoeA, an enzyme in the molybdopterin synthesis pathway, is required for rifamycin SV production in Amycolatopsis mediterranei U32; Wang W et al.; Rifamycin SV contains one amide nitrogen atom at its C(7)N moiety . Earlier labeling studies suggested that nitrogen might be incorporated from a pathway involved in a molybdenum-dependent nitrate reductase . However, no genetic evidence is available thus far . The structural gene moeA, which is involved in molybdopterin synthesis in various organisms, has been cloned from rifamycin SV-producing Amycolatopsis mediterranei strain U32 . The amino acid sequence deduced from the moeA gene showed significant similarity to members of the MoeA protein family and contains all the structural features that are highly conserved in the putative functional domains of MoeA proteins . Southern hybridization showed that there is only one moeA gene in the A . mediterranei genome . To further investigate the possible physiological function of the moeA gene, a double crossover gene replacement was achieved by inserting an aparmycin resistance gene into moeA in the A . mediterranei U32 chromosome . Phenotype analysis showed that the moeA gene is required for A . mediterranei growth in a minimal medium with nitrate as sole nitrogen source, possibly because nitrate reductase activity is diminished due to disruption of the moeA gene . Compared to the wild type strain, moeA-disrupted mutants lost 95% of their rifamycin SV production capacity in complex fermentation media . The results demonstrate that the moeA gene is necessary for rifamycin SV production in A . mediterranei, and that the nitrogen assimilation pathway involved in nitrate reductase is the major pathway for the genesis of the amide nitrogen atom in the rifamycin SV molecule.

Appl Microbiol Biotechnol, 2002 Oct, 60(1-2), 101 - 6 Epub 2002 Jul 13.
Enhancement of pyruvate production by Torulopsis glabrata using a two-stage oxygen supply control strategy; Li Y et al.; The effect of agitation speeds on the performance of producing pyruvate by a multi-vitamin auxotrophic yeast, Torulopsis glabrata, was investigated in batch fermentation . High pyruvate yield on glucose (0.797 g g(-1)) was achieved under high agitation speed (700 rpm), but the glucose consumption rate was rather low (1.14 g l(-1) h(-1)) . Glucose consumption was enhanced under low agitation speed (500 rpm), but the pyruvate yield on glucose decreased to 0.483 g g(-1) . Glycerol production was observed under low agitation speed and decreased with increasing agitation speed . Based on process analysis and carbon flux distribution calculation, a two-stage oxygen supply control strategy was proposed, in which the agitation speed was controlled at 700 rpm in the first 16 h and then switched to 500 rpm . This was experimentally proven to be successful . Relatively high concentration of pyruvate (69.4 g l(-1)), high pyruvate yield on glucose (0.636 g g(-1)), and high glucose consumption rate (1.95 g l(-1)h(-1)) were achieved by applying this strategy . The productivity (1.24 g l(-1) h(-1)) was improved by 36%, 23% and 31%, respectively, compared with fermentations in which agitation speeds were kept constant at 700 rpm, 600 rpm, and 500 rpm . Experimental results indicate that the difference between the performances for producing pyruvate under a favorable state of oxygen supply (dissolved oxygen concentration >50%) was caused by the different regeneration pathways of NADH generated from glycolysis.

Appl Microbiol Biotechnol, 2002 Oct, 60(1-2), 81 - 7 Epub 2002 Aug 24.
Citric acid production by Candida strains under intracellular nitrogen limitation; Anastassiadis S et al.; A suitable strain and important factors influencing citric acid formation in yeasts were identified . Candida oleophila ATCC 20177 was chosen as the best citric acid producer from several Candida strains . Yields of 50 g/l citric acid were produced in shake flask and 80 g/l in fed-batch fermentations with 1.5 and 3 g/l NH(4)Cl under non-optimized conditions . Ammonium nitrogen was identified as the limiting substrate for citrate formation . Citric acid excretion begins a few hours after exhaustion of nitrogen in the medium . The importance of intracellular nitrogen limitation was clarified by elemental analysis of C . oleophila biomass . The nitrogen content of C . oleophila biomass decreased from 7.45% during the growth phase to 3.96% in the production phase . The biomass contained less carbon and more trace elements in the growth phase compared with the production phase . Relatively high intracellular NH(4)(+) concentration of about 1.2 mg/g biomass (~37.4 mM) was found during the production phase . The low intracellular nitrogen content and increase of intracellular NH(4)(+) concentration, possibly caused by proteolysis following extracellular nitrogen exhaustion, trigger citric acid production . Intracellular nitrogen limitation and the increase in intracellular NH(4)(+) concentration are the most important factors influencing citric acid formation in yeasts.

Appl Microbiol Biotechnol, 2002 Oct, 60(1-2), 67 - 72 Epub 2002 Aug 28.
Improving ethanol production and viability of Saccharomyces cerevisiae by a vitamin feeding strategy during fed-batch process; Alfenore S et al.; Several bottlenecks in the alcoholic fermentation process must be overcome to reach a very high and competitive performance of bioethanol production by the yeast Saccharomyces cerevisiae . In this paper, a nutritional strategy is described that allowed S . cerevisiae to produce a final ethanol titre of 19% (v/v) ethanol in 45 h in a fed-batch culture at 30 degrees C . This performance was achieved by implementing exponential feeding of vitamins throughout the fermentation process . In comparison to an initial addition of a vitamin cocktail, an increase in the amount of vitamins and an exponential vitamin feeding strategy improved the final ethanol titre from 126 g l(-1) to 135 g l(-1) and 147 g l(-1), respectively . A maximum instantaneous productivity of 9.5 g l(-1) h(-1) was reached in the best fermentation . These performances resulted from improvements in growth, the specific ethanol production rate, and the concentration of viable cells in response to the nutritional strategy.

J Agric Food Chem, 2002 Oct 23, 50(22), 6557 - 62
Impact of whole wheat flour and its milling fractions on the cecal fermentations and the plasma and liver lipids in rats; Adam A et al.; The aim of the present work was to evaluate the cholesterol-lowering potency of the different milling fractions of whole wheat flour, by investigating the effects of these wheat fractions (white flour, whole flour, and bran) on digestive fermentations and lipid metabolism in Wistar rats . Compared to the control, which was fiber-free, the different cereal fractions did not affect the daily food intake or weight gain . The white flour and whole flour diets markedly enlarged the cecum and elicited acidic fermentations (pH approximately 6.2), whereas bran was less effective . It appears that white flour rather promoted propionate-rich fermentations (+62%), whereas bran favored butyrate-rich fermentations (+178%) . White flour or bran did not significantly affect total steroid excretion, but whole flour was effective (+41%) . Both white flour and whole flour decreased cholesterol in the d < 1.040 fraction, but only whole flour significantly lowered cholesterolemia . However, all the cereal diets significantly decreased liver lipids, whole flour being the most potent (-54%) . In conclusion, the totality of the wheat grain is important for cholesterol- and triglyceride-lowering effects, and the splitting up of the grain alters its health effects.

J Agric Food Chem, 2002 Oct 23, 50(22), 6501 - 3
Isolation and identification of DPPH radical scavenging compounds in Kurosu (Japanese unpolished rice vinegar); Shimoji Y et al.; Dihydroferulic acid (DFA) and dihydrosinapic acid (DSA) were isolated from Kurosu (unpolished rice vinegar) as the major constituents responsible for Kurosu's radical scavenging activity . The levels of antioxidative activity of DFA and DSA in DPPH radical scavenging were higher than those of their respective structurally related compounds, ferulic acid and sinapic acid . The concentrations of DFA and DSA were low in common rice vinegar (polished rice vinegar), suggesting that Kurosu is more advantageous than rice vinegars as an antioxidative food item . As the concentrations of DFA and DSA were low in unpolished rice, too, these acids are thought to be produced in Kurosu through the process of the fermentation from ferulic acid and sinapic acid, respectively.

J Food Prot, 2002 Oct, 65(10), 1541 - 4
Viability of Escherichia coli O157:H7 during manufacturing and storage of a fermented, semidry soudjouk-style sausage; Calicioglu M et al.; Soudjouk-style batter was inoculated with a five-strain mixture of Escherichia coli O157:H7 at about 7.6 log10 CFU/g in each of two trials . The sticks were fermented and dried at 22 degrees C and 50% relative humidity (RH) for 3 days and then at 9 degrees C and 40% RH for 18 h . After being flattened to about 1.25 cm, the sticks were conditioned at 38 degrees C and 70% RH or at 22 degrees C and 50% RH for about 3 days . After the latter conditioning treatment, sticks either were cooked to an internal temperature of 63 degrees C or received no heat treatment . Final mean pH values after conditioning at 22 degrees C and 50% RH for soudjouk manufactured with a starter culture and dextrose (1.0%) and for soudjouk manufactured without a starter culture were about 4.9 and 6.0, respectively . For soudjouk produced with a starter culture, pathogen numbers were reduced by 4.53 and 0.88 log10 CFU/g after conditioning at 38 degrees C and 70% RH and at 22 degrees C and 50% RH, respectively . For soudjouk produced via natural fermentation, pathogen numbers were reduced by 1.39 and 0.09 log10 CFU/g after conditioning at 38 degrees C and 70% RH and at 22 degrees C and 50% RH, respectively . Cooking reduced pathogen numbers to below the levels detectable by direct plating (<1.0 log10 CFU/g) and by enrichment for soudjouk produced with a starter culture and also reduced pathogen numbers by 6.28 log10 CFU/g for soudjouk produced via natural fermentation . However, cooking also resulted in an unacceptable product . In general, the reduction in pathogen numbers achieved by storage at ambient temperature (25 degrees C) was greater than that achieved by storage at cooler temperatures (4 and 15 degrees C), particularly for soudjouk prepared with a starter culture (for which a final pH value of 4.8 and a 6.4-log10 reduction were obtained after 21 days at 25 degrees C) rather than for that prepared without a starter culture (for which a final pH value of 6.1 and a 2.6-log10 reduction were obtained after 21 days at 25 degrees C) . These results indicate that naturally fermented old-country-type sausage may allow the survival of E . coli O157:H7 in the absence of controlled fermentation, postfermentation cooking, and/or an ambient-storage processing step.

Ann Pharm Fr, 2002 Sep, 60(5), 333 - 40
{Certification of suitability of monographs of the European pharmacopoeia}; Artiges A; In April 1994, t he European Pharmacopoeia Commission set up a new procedure for the "Certification of Suitability of Monographs of the European Pharmacopoela" to deal with the changing requirements of the licensing authorities and the growth of international trade, notably as regards raw materials to be used in the manufacture of medicines . This procedure was the result of extensive collaboration between the licensing authorities of the European Union and the other parties to the European Pharmacopoeia Convention, and it enables a manufacturer of a raw material for pharmaceutical use to demonstrate that the purity of their substance is suitably controlled by the monograph of the European Pharmacopoeia; this demonstration is now required by the guideline on "Requirements in Relation to Active Substances" published in Volume 3A of the Rules Governing Medicinal Products in the European Union . Initially set up in response to problems with impurities of synthesis and residual solvents that can vary from one manufacturer to another, the procedure has gradually been extended to a very wide range of products including products of fermentation and, more recently, products with risk of transmitting agents of animal spongiform encephalopathies.

J Bacteriol, 2002 Nov, 184(21), 5903 - 11
Carbon monoxide cycling by Desulfovibrio vulgaris Hildenborough; Voordouw G; Sulfate-reducing bacteria, like Desulfovibrio vulgaris Hildenborough, use the reduction of sulfate as a sink for electrons liberated in oxidation reactions of organic substrates . The rate of the latter exceeds that of sulfate reduction at the onset of growth, causing a temporary accumulation of hydrogen and other fermentation products (the hydrogen or fermentation burst) . In addition to hydrogen, D . vulgaris was found to produce significant amounts of carbon monoxide during the fermentation burst . With excess sulfate, the hyd mutant (lacking periplasmic Fe-only hydrogenase) and hmc mutant (lacking the membrane-bound, electron-transporting Hmc complex) strains produced increased amounts of hydrogen from lactate and formate compared to wild-type D . vulgaris during the fermentation burst . Both hydrogen and CO were produced from pyruvate, with the hyd mutant producing the largest transient amounts of CO . When grown with lactate and excess sulfate, the hyd mutant also exhibited a temporary pause in sulfate reduction at the start of stationary phase, resulting in production of 600 ppm of headspace hydrogen and 6,000 ppm of CO, which disappeared when sulfate reduction resumed . Cultures with an excess of the organic electron donor showed production of large amounts of hydrogen, but no CO, from lactate . Pyruvate fermentation was diverse, with the hmc mutant producing 75,000 ppm of hydrogen, the hyd mutant producing 4,000 ppm of CO, and the wild-type strain producing no significant amount of either as a fermentation end product . The wild type was most active in transient production of an organic acid intermediate, tentatively identified as fumarate, indicating increased formation of organic fermentation end products in the wild-type strain . These results suggest that alternative routes for pyruvate fermentation resulting in production of hydrogen or CO exist in D . vulgaris . The CO produced can be reoxidized through a CO dehydrogenase, the presence of which is indicated in the genome sequence.

J AOAC Int, 2002 Sep-Oct, 85(5), 1052 - 6
N-nitroso compounds and mutagens in Chinese fermented (sour) corn pancakes; Groves FD et al.; Stomach cancer rates in rural Linqu County, Shandong Province, China, are exceptionally high . A previous case-control study revealed that the risk of stomach cancer was 30% higher among those who consumed sour (fermented) corn pancakes at least daily . A previous study of the sour pancakes reported volatile nitrosamines in most specimens, and almost half reportedly showed mutagenic activity . Few households currently consume sour pancakes, and the duration of fermentation has been shortened . We tested specimens of pancake batter and sour pancakes from Linqu County for mutagenic activity using the Ames test; for N-nitroso compounds (NOC) we used the Nitrolite-thermal energy analysis (TEA) method . Results of the Ames test were inconclusive: only 1 out of 15 cooked pancakes showed a positive mutagenic response, and all 15 batter specimens were negative; however, several batter specimens showed a weakly positive trend of mutagenicity with extract concentration . Our assay for total nitroso compounds was weakly positive in only 1 out of 15 specimens of sour pancake batter . That specimen was also tested by gas chromatography-TEA for nitrosaminoacids and volatile nitrosamines, but none were detected . It seems unlikely that the Chinese sour pancakes are significantly contaminated by NOC or other mutagens.

J Antibiot (Tokyo), 2002 Aug, 55(8), 702 - 6
New dithiolopyrrolone antibiotics from Saccharothrix sp . SA 233 . II . Physicochemical properties and structure elucidation; Lamari L et al.; Three new natural dithiopyrrolone antibiotics, 3-methyl-2-butenoylpyrrothine (1), tigloylpyrrothine (2), and n-butyropyrrothine (3) were isolated along with the known isobutyropyrrothine (4) and thiolutin (5) from the fermentation broth of Saccharothrix sp . SA 233 . The structures of the novel compounds were established on the basis on their spectral data.

J Antibiot (Tokyo), 2002 Aug, 55(8), 693 - 5
Aspergillin PZ, a novel isoindole-alkaloid from Aspergillus awamori; Zhang Y et al.; Aspergillin PZ was obtained from the fermentation of Aspergillus awamori (Nakazawa) by activity-guided fractionation and purification . Its structure was elucidated on the basis of spectral data, especially by 2D NMR, and finally confirmed by an X-ray analysis . It could induce conidia of P . oryzae to deform moderately.

J Antibiot (Tokyo), 2002 Aug, 55(8), 685 - 92
TMC-256A1 and C1, new inhibitors of IL-4 signal transduction produced by Aspergillus niger var niger TC 1629; Sakurai M et al.; New inhibitors of IL-4 signal transduction, designated as TMC-256A1 and C1, were discovered together with TMC-256B1, a previously known dihydronaphthopyrone, from the fermentation broth of Aspergillus niger var niger TC 1629 by using an IL-4 driven reporter gene assay . Based on spectroscopic analyses, TMC-256A1 and C1 were found to be new members of the naphthopyrone antibiotics . TMC-256A1, B1 and C1 inhibited the IL-4 driven luciferase activity with IC50 values of 25 microM, 30 microM and 1.7 microM, respectively in this assay system . Furthermore, these compounds inhibited the expression of germline C epsilon mRNA with IC50 values of 6.6 microM , 34 microM and 0.31 microM, respectively.

J Chem Ecol, 2002 Aug, 28(8), 1527 - 47
Semiochemical-mediated flight responses of sap beetle vectors of oak wilt, Ceratocystis fagacearum; Kyhl JF et al.; The sap beetle, Colopterus truncatus (Coleoptera: Nitidulidae), is one of the primary vectors of the oak wilt pathogen, Ceratocystis fagacearum, in the north-central United States . Field behavioral assays utilizing various release rates and blends of three methyl-branched hydrocarbon aggregation pheromone components showed that flight responses of this beetle were similar in Illinois and Minnesota populations . In both locations, both sexes of the beetle responded synergistically to a combination of the three-component pheromone and fermenting whole-wheat bread dough . Further, Colopterus truncatus preferred a high release rate over a low release rate of the three-component blend . In both locations, the response of C . truncatus to a simplified version of the pheromone consisting of (2E,4E,6E)-3,5-dimethyl-2,4,6-octatriene (1) and (2E,4E,6E,8E)-3,5,7-trimethyl-2,4,6,8-decatetraene (3) was not significantly different from the response to the three-component blend . An experiment in Illinois with all possible combinations of the components demonstrated that the decatetraene (3) was the crucial component in the blend; of all treatments, the maximal response was elicited by 3 + dough . Chipped bark, phloem, and xylem from northern pin oak, Quercus ellipsoidalis, was not attractive to C . truncatus in Minnesota . During a weekly survey over two seasons in Minnesota, C . truncatus flew in response to the three-component pheromone between early April and early July, with the maximum responses coming on May 4, 2000 and April 20, 2001 . During both years, more than 98% of the beetles were trapped between April 14 and June 1 . During the same survey, Glischrochilus spp . (Nitiduildae) flew during longer periods of the summer, particularly in 2001 . The sex ratio of C . truncatus responding during all experiments was female-biased (1.8:1, female-male), which is characteristic of other male-produced coleopteran aggregation pheromones . Other sap beetles that play a minor role in the pathobiology of C . fagacearum also responded in experiments conducted in Minnesota . Carpophilus brachypterus Say was cross-attracted to the two- and three-component blends of the C . truncatus pheromone and dough, whereas two Glischrochilus spp . were attracted to all treatments that contained dough.

Acta Biol Hung, 2002, 53(3), 389 - 401
Selection of strain and optimization of mutanase production in submerged cultures of Trichoderma harzianum; Wiater A et al.; Nineteen fungal strains belonging to different genera were tested for extracellular mutanase production in shaken flasks . The optimal enzymatic activity was achieved by Trichoderma harzianum F-470, a strain for which the mutanase productivity has not yet been published . Some of factors affecting the enzyme production in shaken flasks and aerated fermenter cultures have been standardized . Mandels mineral medium with initial pH 5.3, containing 0.25% mutan and inoculated with 10% of the 48-h mycelium, was the best for enzyme production . A slight mutanolytic activity was also found when sucrose, raffinose, lactose and melibiose were carbon sources . Application of optimized medium and cultural conditions, as well as use of a fermenter with automatic pH control set at pH 6.0 enabled to obtain a high mutanase yield (0.33 U/ml, 2.5 U/mg protein) in a short time (2-3 days) . The enzyme in crude state was stable over a pH range of 4.5-6.0, and at temperatures up to 35 degrees C; its maximum activity was at 40 degrees C and at pH 5.5.

Proc Natl Acad Sci U S A, 2002 Oct 15, 99(21), 13431 - 6 Epub 2002 Oct 07.
The Ume6 regulon coordinates metabolic and meiotic gene expression in yeast; Williams RM et al.; The Ume6 transcription factor in yeast is known to both repress and activate expression of diverse genes during growth and meiotic development . To obtain a more complete profile of the functions regulated by this protein, microarray analysis was used to examine transcription in wild-type and ume6Delta diploids during vegetative growth in glucose and acetate . Two different genetic backgrounds (W303 and SK1) were examined to identify a core set of strain-independent Ume6-regulated genes . Among genes whose expression is controlled by Ume6 in both backgrounds, 82 contain homologies to the Ume6-binding site (URS1) and are expected to be directly regulated by Ume6 . The vast majority of those whose functions are known participate in carbon/nitrogen metabolism and/or meiosis . Approximately half of the Ume6 direct targets are induced during meiosis, with most falling into the early meiotic expression class (cluster 4), and a smaller subset in the middle and later classes (clusters 5-7) . Based on these data, we propose that Ume6 serves a unique role in diploid cells, coupling metabolic responses to nutritional cues with the initiation and progression of meiosis . Finally, expression patterns in the two genetic backgrounds suggest that SK1 is better adapted to respiration and W303 to fermentation, which may in part account for the more efficient and synchronous sporulation of SK1.

J Environ Sci Health A Tox Hazard Subst Environ Eng, 2002 Sep, 37(8), 1533 - 43
Domestic wastewater biosolids accumulation by liquid state bioconversion process for rapid composting; Fakhru'l-Razi A et al.; Bioconversion of higher strength of domestic wastewater biosolids (sludge) (4% w/w of TSS) by mixed fungal culture of Aspergillus niger and Penicillium corylophilum was studied in a laboratory . The effect of potential mixed fungi on domestic wastewater sludge accelerated the liquid state bioconversion (LSB) process . The highest production of dry sludge cake (biosolids) was enriched with fungal biomass to about 85.66 g/kg containing 25.23 g/kg of protein after 8 days of treatment . The results presented in this study revealed that the reduction of chemical oxygen demand (COD), total suspended solid (TSS), and specific resistance to filtration (SRF) of treated sludge were highly influenced by the fungal culture as compared to control (uninnoculated) . The maximum removal rates in treated sludge (biosolids) supernatant recorded were 92% of COD and 98.8% of TSS . Lower SRF (1.08 x 10(12) m/kg) was perceived in microbially treated sludge after 6 days of fermentation . The observed parameters were highly influenced after 8 days of treatment . The influence of pH was also studied and presented in the paper.

Antonie Van Leeuwenhoek, 2002 Aug, 82(1-4), 303 - 21
Bacteriophage-resistance systems in dairy starter strains: molecular analysis to application; Coffey A et al.; Starter inhibition by bacteriophage infection in dairy fermentations can limit the usage of specific bacterial strains used in the manufacture of Cheddar, Mozzarella and other cheeses and can result in substantial economic losses . A variety of practical measures to alleviate the problem of phage infection have been adopted over the years but has invariably resulted in a very limited number of strains which can withstand intensive usage in industry . The application of genetic techniques to improve the phage-resistance of starter cultures for dairy fermentations has been intensively studied for the last 20 years to a point where this approach now has significant potential to alleviate the problem . This paper highlights the recent findings and developments that have been described in the literature that will have an impact on improvement of the phage-resistance of starter cultures.

Biotechnol Prog, 2002 Sep-Oct, 18(5), 1130 - 2
High-cell-density fermentation of recombinant Saccharomyces cerevisiae using glycerol; Eugene Raj A et al.; To obtain a high cell density of recombinant Saccharomyces cerevisiae (INVSc 1 strain bearing a 2 microm plasmid, pYES2 containing a GAL1 promoter for expression of the beta-galactosidase gene), the yeast was grown with glycerol as the substrate by fed-batch fermentation . The feeding strategy was based on an on-line response of the medium pH to the consumption of glycerol . The approach was to feed excess carbon into the medium to create a benign environment for rapid biomass buildup . During cell growth in the presence of glycerol, the release of protons in the medium caused a decrease in pH and the consumption rate of ammonium phosphate served as an on-line indicator for the metabolic rate of the organism . The extent of glycerol feeding in a fed-batch mode with pH control at 5.0 +/- 0.1 was ascertained from the automatic addition of ammonium phosphate to the medium . The glycerol feeding to ammonium phosphate addition ratio was found to be 2.5-3.0 . On the basis of the experiments, a maximum dry cell biomass of 140 g per liter and a productivity of 5.5 g DCW/L/h were achieved . The high cell density of S . cerevisiae obtained with good plasmid stability suggested a simple and efficient fermentation protocol for recombinant protein production.

Biotechnol Prog, 2002 Sep-Oct, 18(5), 1047 - 53
Dual excitation ratiometric fluorescent pH sensor for noninvasive bioprocess monitoring: development and application; Kermis HR et al.; The development and application of a fluorescent excitation-ratiometric, noninvasive pH sensor for continuous on-line fermentation monitoring is presented . The ratiometric approach is robust and insensitive to factors such as source intensity, photobleaching, or orientation of the patch, and since measurements can be made with external instrumentation and without direct contact with the patch, detection is completely noninvasive . The fluorescent dye 8-hydroxy-1,3,6-pyrene trisulfonic acid was immobilized onto Dowex strongly basic anion-exchange resin, which was subsequently entrapped into a proton-permeable hydrogel layer . The sensor layer was polymerized directly onto a white microfiltration membrane backing that provided an optical barrier to the fluorescence and scatter of the fermentation medium . The ratio of emission intensity at 515 nm excited at 468 nm to that excited at 408 nm correlated well with the pH of clear buffers, over the pH range of 6-9 . The sensor responded rapidly (<9 min) and reversibly to changes in the solution pH with high precision . The sterilizable HPTS sensor was used for on-line pH monitoring of an E . coli fermentation . The output from the indwelling sensor patch was always in good agreement with the pH recorded off-line with an ISFET probe, with a maximum discrepancy of 0.05 pH units . The sensor is easily adaptable to closed-loop feedback control systems.

Biotechnol Prog, 2002 Sep-Oct, 18(5), 935 - 41
Pulse experiments as a prerequisite for the quantification of in vivo enzyme kinetics in aromatic amino acid pathway of Escherichia coli; Schmitz M et al.; Glucose pulse experiments were performed to elucidate their effects on the carbon flux into the aromatic amino acid pathway in different Escherichia coli strains . Using a 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP, aroB(-))-producing strain, a fed-batch fermentation strategy specialized for glucose pulse experiments was developed and further applied for 3-dehydroshikimate (DHS, aroE(-))- and shikimate 3-phosphate (S3P, aroA(-))-producing E . coli strains . The strains overexpress a feedback-resistant DAHP synthase and additional enzymes to prevent rate-limiting steps in the aromatic amino acid pathway . Changes of carbon flux into the aromatic amino acid pathway were determined via extracellular metabolite accumulations using (1)H NMR and HPLC measurements . As an important result, a close relationship between pulse intensity and aromatic metabolite formation rates was identified . The more downstream an aromatic pathway intermediate was located, the stronger the glucose pulse intensity had to be in order to detect significant changes in product formation . However, with the experimental conditions chosen, changes after pulse were detected even for shikimate 3-phosphate, the most downstream accumulating metabolite of this experimental series . Hence glucose pulse experiments are assumed to be a promising tool even for the analysis of final pathway products such as, for example, L-phenylalanine.

Biotechnol Prog, 2002 Sep-Oct, 18(5), 921 - 6
Microbial hydrogen production with immobilized sewage sludge; Wu SY et al.; Municipal sewage sludge was immobilized to produce hydrogen gas under anaerobic conditions . Cell immobilization was essentially achieved by gel entrapment approaches, which were physically or chemically modified by addition of activated carbon (AC), polyurethane (PU), and acrylic latex plus silicone (ALSC) . The performance of hydrogen fermentation with a variety of immobilized-cell systems was assessed to identify the optimal type of immobilized cells for practical uses . With sucrose as the limiting carbon source, hydrogen production was more efficient with the immobilized-cell system than with the suspended-cell system, and in both cases the predominant soluble metabolites were butyric acid and acetic acid . Addition of activated carbon into alginate gel (denoted as CA/AC cells) enhanced the hydrogen production rate (v(H2)) and substrate-based yield (Y((H2)/sucrose)) by 70% and 52%, respectively, over the conventional alginate-immobilized cells . Further supplementation of polyurethane or acrylic latex/silicone increased the mechanical strength and operation stability of the immobilized cells but caused a decrease in the hydrogen production rate . Kinetic studies show that the dependence of specific hydrogen production rates on the concentration of limiting substrate (sucrose) can be described by Michaelis-Menten model with good agreement . The kinetic analysis suggests that CA/AC cells may contain higher concentration of active biocatalysts for hydrogen production, while PU and ALSC cells had better affinity to the substrate . Acclimation of the immobilized cells led to a remarkable enhancement in v(H2) with a 25-fold increase for CA/AC and ca . 10- to 15-fold increases for PU and ALSC cells . However, the ALSC cells were found to have better durability than PU and CA/AC cells as they allowed stable hydrogen production for over 24 repeated runs.

Food Nutr Bull, 2002 Sep, 23(3 Suppl), 241 - 5
Effect of traditional fermentation and malting on phytic acid and mineral availability from sorghum (Sorghum bicolor) and finger millet (Eleusine coracana) grain varieties grown in Kenya; Makokha AO et al.; Sorghum and finger millet grains are traditional staple foods in Kenya . However, they have naturally occurring anti-nutritional factors, such as phytic acid, that decrease their dietary availability . This work determined the effect of fermentation and malting on the phytic acid content of, and mineral availability in five varieties of sorghum and four varieties of finger millet grain grown in Kenya . Phytic acid ranged from 875.1 to 2,211.9 mg/100 g in sorghum . The levels in finger millet ranged from 851.6 to 1,419.4 mg/100 g grain . Fermentation resulted in a mean decrease of phytic acid in of 64.8% after 96 hours and 39.0% after 72 hours in sorghum grain . In finger millet, there was a mean decrease of 72.3% and 54.3% after 96 and 72 hours, respectively . Malting also resulted in a mean decrease of 23.9 and 45.3% after 72 and 96 hours, respectively . The extent of decrease of phytic acid differed among the grain varieties . Fermentation increased the rate of available iron, manganese, and calcium in both sorghum and finger millet . The available minerals were generally higher in finger millet than in sorghum after fermentation . Fermentation was also more effective than malting in reducing phytic acid in sorghum and finger millet . Simple traditional food processing methods can therefore be used to increase mineral availability.

J Dairy Sci, 2002 Sep, 85(9), 2290 - 8
Effects of day relative to parturition and dietary crude protein on rumen fermentation in prepartum transition cows; Dorshorst ME et al.; Twelve Holstein cows and eight heifers were used in a randomized block design to examine changes in rumen fermentation and blood metabolites as animals fed diets containing 9.1 (low protein, LP) or 11.2% crude protein (moderate protein, MP) approached parturition . Animals were blocked by expected calving date and parity . Diets were isocaloric (1.58 Mcal/kg of dry matter), and the difference in dietary crude protein was achieved by the addition of urea . Diets were fed as total mixed rations for ad libitum intake . Dry matter intake decreased from 12.2 kg/d on d -21 to 9.1 kg/d on d -1 relative to parturition and was unaffected by dietary crude protein or parity . Rumen ammonia concentrations were higher for animals fed the MP diet compared with those fed the LP diet (1.8 vs . 3.7 mM) . In situ neutral detergent fiber disappearance was not affected by dietary crude protein after 8 h of incubation, but it did decrease from 25.8% on d -21 to 23.3% on d -1 . After 16, 24, and 48 h of incubation, neutral detergent fiber disappearance was greater for animals consuming MP than for animals consuming LP (30.6 vs . 34.4%, 37.0 vs . 44.1%, and 57.9 vs . 65.1%, respectively) . There was a parity x treatment interaction after 48 h of incubation . Fiber disappearance was 55.7 or 65.9% for heifers and 60.0 or 64.7% for cows fed LP or MP, respectively . The solids rate of passage was not affected by treatment, parity, or day relative to parturition . Concentrations of plasma nonesterified fatty acid (NEFA) increased from 218 microEq/L on d -21 to 619 microEq/L on d -1 . There was an interaction between day relative to parturition and parity on NEFA concentrations . Heifers had plasma NEFA concentrations of 272 and 772 microEq/L on d -21 and -1, respectively . Cows had NEFA concentrations of 164 and 467 microEq/L on d -21 and -1, respectively . Plasma glucose concentrations were affected by parity (65.8 and 58.8 mg/dl for heifers and cows) . Plasma urea, NEFA, beta-hydroxybutyrate, and glucose concentrations were not affected by dietary treatment . In conclusion, heifers tended to have higher plasma concentrations of NEFA and glucose, but did not have elevated concentrations of beta-hydroxybutyrate . Rumen fermentation was relatively unaffected by day relative to parturition, but feeding less than 11% crude protein to prefresh transition cows may limit ruminal fiber degradation.

J Basic Microbiol, 2002, 42(5), 320 - 6
Synthesis of alpha-amylase by Aspergillus oryzae in solid-state fermentation; Francis F et al.; Spent Brewing Grains (SBG) was evaluated for its efficacy to be used as sole carbon source for the synthesis of alpha-amylase in solid-state fermentation using a fungal strain of Aspergillus oryzae NRRL 6270 . Enzyme production was superior when the culture grew on mesophilic temperatures and best yields were at 25 degrees C . At 30 degrees C, yields were almost comparable . Maximum production of alpha-amylase {6870 U/g dry substrate (gds)} was obtained when SSF was carried out at 30 degrees C for 96 h using SBG medium, which had initial moisture of 70% and was inoculated using a spore suspension containing 1 x 10(7) spores/ml . Supplementation of SBG with external carbon sources such as mono-, di and polysaccharides caused repression in enzyme synthesis by the fungal culture.

Int J Syst Evol Microbiol, 2002 Sep, 52(Pt 5), 1851 - 7
Rheinheimera baltica gen . nov., sp . nov., a blue-coloured bacterium isolated from the central Baltic Sea; Brettar I et al.; A set of taxonomically unique, blue-coloured bacterial isolates are described on the basis of physiological and biochemical characterization, fatty acid profiling and analyses of 16S rDNA sequences . The flagellated, non-fermentative strains were isolated in 1986, 1987 and 1998 from different layers of the water column of the central Baltic Sea . According to 16S rDNA sequences, all strains are very closely related to each other and to strains from several other marine environments, including the deep sea . Thus, the described species seems to be widespread in marine habitats . According to DNA-DNA hybridization, the strains described can be considered to belong to the same species . The bacteria grew at temperatures from 4 to 30 degrees C, with an optimum around 20-25 degrees C . Growth was observed at salinities from 0 to 30, with an optimum between 10 and 30 and no growth at high salinities . The dominant fatty acids were 16:1omega7c, 16:0 and 18:1omega7c . The G+C content of the DNA ranged from 47.8 to 48.9 mol% . Phylogenetic analyses of the 16S rDNA sequences revealed a clear affiliation with members of the gamma-Proteobacteria . The closest relationship was seen with Alishewanella fetalis but, in terms of physiology, colour and fatty acids, the bacteria described are rather distant from A . fetalis . To honour the marine microbiologist Gerhard Rheinheimer, the name Rheinheimera baltica gen . nov., sp . nov., is suggested for the Baltic isolates, with the type strain OSBAC1T (= DSM 14885T = LMG 21511T).

Lett Appl Microbiol, 2002, 35(4), 338 - 42
Screening of pyruvate-producing yeast and effect of nutritional conditions on pyruvate production; Wang Q et al.; AIMS: To find a yeast strain that can overproduce pyruvate and to investigate the effect of nutrients on pyruvate production . METHODS AND RESULTS: Trichosporon cutaneum PD70, a yeast strain that can overproduce pyruvate, was isolated from shake-flask cultures of 132 yeast strains . Pyruvate was measured by the HPLC or DNP method (see Materials and methods) . Pyruvate production reached approximately 30.0 +/- 1.0 g l(-1) in basal fermentation medium . Different nutrient supplements had great effects on pyruvate production . Some of the conditions that gave the highest yield are described . CONCLUSIONS: Exogenous thiamine supplement caused a decrease in pyruvate yield . Some amino acids, such as L-arginine, L-isoleucine and L-valine, caused a minor increase in pyruvate yield . Soybean peptone was the most suitable nitrogen source for pyruvate production . A glucose concentration of 15% in fermentation medium gave the highest yield (34.6 g l(-1)) and the highest yield against consumed glucose (0.429 g g(-1)) . SIGNIFICANCE AND IMPACT OF THE STUDY: Nutrients have significant impacts on pyruvate production . As a pyruvate overproducing yeast strain independent of exogenous vitamins or amino acids, T . cutaneum PD70 provides an advantage for commercial pyruvate production.

Lett Appl Microbiol, 2002, 35(4), 305 - 10
Identification and characterization of Saccharomyces cerevisiae and Saccharomyces paradoxus strains isolated from Croatian vineyards; Redzepovic S et al.; AIMS: The identification, differentiation and characterization of indigenous Saccharomyces sensu stricto strains isolated from Croatian vineyards and the evaluation of their oenological potential . METHODS AND RESULTS: A total of 47 Saccharomyces sensu stricto strains were isolated from Chardonnay grapes and identified by physiological and molecular genetic methods . By using the standard physiological and biochemical tests, six isolates were identified as Saccharomyces cerevisiae and 41 as Saccharomyces paradoxus . However, PCR-RFLP analyses of the internal transcribed spacer (ITS1) region of the 18S ribosomal DNA identified 12 of the isolates as S.cerevisiae and 35 as S . paradoxus . Fermentation trials in a grape juice medium showed that these isolates ferment vigorously at 18 degrees C and display tolerance to high levels of ethanol . None of these isolates appeared to produce either hydrogen sulphide or killer toxins . CONCLUSION: Saccharomyces paradoxus, possessing potentially important oenological characteristics, occurs in much higher numbers than S . cerevisiae in the indigenous population of Saccharomyces sensu stricto strains in Croatian vineyards . SIGNIFICANCE AND IMPACT OF THE STUDY: This study forms an essential step towards the preservation and exploitation of the hidden oenological potential of the untapped wealth of yeast biodiversity in the Croatian grape-growing regions . The results obtained demonstrate the value of using molecular genetic methods, such as PCR-RFLP analyses, in conjunction with the traditional taxonomic methods based on phenotypic characteristics in such ecotaxonomic surveys . The results also shed some light on the ecology and oenological potential of S.paradoxus, which is considered to be the natural parent species of the domesticated species of the Saccharomyces sensu stricto group.

J Agric Food Chem, 2002 Oct 9, 50(21), 6090 - 6
Application of a DNA analysis method for the cultivar identification of grape musts and experimental and commercial wines of Vitis vinifera L . using microsatellite markers; Garcia-Beneytez E et al.; A DNA-based method has been applied to the identification of several musts and wines using microsatellite markers . DNA was extracted from the solid phases of sixteen monovarietal and five multivarietal musts (mixtures of two musts down to a 4:1 proportion) and they were genotyped at seven microsatellites through a multiplex PCR reaction and automated fluorescent detection . PCR multiplexing was successful in monovarietal musts, but should be used with caution with at least some markers and in multivarietal musts . The same extraction and detection methods were unsuccessfully applied to the solid and liquid phases of five monovarietal commercial wines, even after using different concentration procedures . Nucleic acids presence was then studied in a recent must, during the fermentation process, and during the subsequent steps of winemaking . Genotyping was possible in the resulting experimental wine until decanting, when the particles in suspension were removed . These results suggest that wine authentication through DNA analysis is not possible in commercial wines, in the tested conditions.

J Agric Food Chem, 2002 Oct 9, 50(21), 6078 - 85
Localization of puroindoline-a and lipids in bread dough using confocal scanning laser microscopy; Dubreil L et al.; Puroindolines are lipid-binding proteins from wheat flour that play a significant role in bread crumb texture . The localization of wheat flour lipids and puroindoline-a (PIN-a) in bread dough was studied by confocal scanning laser microscopy (CSLM) . Wheat lipids were located around gas cells (GC) and embedded within the protein-starch matrix (SPM) of the dough . PIN-a was mainly located in the matrix of dough, where it was associated with lipids . In contrast, in defatted dough, PIN-a was found around GC . Addition of puroindolines in bread dough induced a defatting of the gas bubble surface and a decrease of the lipid vesicles and/or droplet size embedded within the SPM . Therefore, puroindolines control the lipid partitioning within the different phases of dough, a phenomenon that should have important consequence on the gas bubble expansion and GC formation in the further stages (fermentation, baking) of the bread-making process.

J Agric Food Chem, 2002 Oct 9, 50(21), 5932 - 8
Controlled-atmosphere effects on postharvest quality and antioxidant activity of cranberry fruits; Gunes G et al.; The effects of controlled-atmosphere (CA) storage on the firmness, respiration rate, quality, weight loss, total phenolics and flavonoids contents, and total antioxidant activities of the Pilgrim and Stevens cultivars of cranberries (Vaccinium macrocarpon Aiton) have been studied during storage in atmospheres of 2, 21, and 70% O(2) with 0, 15, and 30% CO(2) (balance N(2)); and 100% N(2) at 3 degrees C . Elevated CO(2) concentrations decreased bruising, physiological breakdown, and decay of berries, thereby reducing fruit losses . Respiration and weight loss of fruits decreased, but fruit softening increased, at higher CO(2) concentrations . Accumulations of acetaldehyde, ethanol, and ethyl acetate varied by cultivar and storage atmosphere but were generally highest in the 2 and 70% O(2) and 100% N(2) atmospheres and increased in response to elevated CO(2) concentrations . Overall, the 30% CO(2) plus 21% O(2) atmosphere appeared optimal for the storage of cranberries . Sensory analysis is required, however, to confirm that accumulations of fermentation products at this atmosphere are acceptable for consumers . Stevens fruits had a higher phenolics content and total antioxidant activity than Pilgrim fruits . The storage atmosphere did not affect the content of total phenolics or flavonoids . However, the total antioxidant activity of the fruits increased overall by about 45% in fruits stored in air . This increase was prevented by storage in 30% CO(2) plus 21% O(2).

Protein Expr Purif, 2002 Oct, 26(1), 139 - 48
Recombinant insect cell expression and purification of human beta-secretase (BACE-1) for X-ray crystallography; Bruinzeel W et al.; Human beta-secretase (BACE-1) is a type I integral membrane aspartic protease that catalyzes the internal cleavage of the amyloid precursor protein (APP), generating the N-terminus of the Abeta peptide . The generation and subsequent extracellular deposition of Abeta(1-42) peptide into amyloid plaques in the brain constitute one of the hallmarks of Alzheimer's disease (AD), a common debilitating neurodegenerative disorder . Inhibition of BACE-1 is considered an excellent therapeutic strategy against AD . To generate pure enzyme for protein crystallography and subsequent structure-based drug design, we have expressed a soluble, unglycosylated, 6xHis-tagged form of proBACE-1 in insect cells using baculovirus infection . To avoid production of a mixture of the pro-enzyme form and the mature form of BACE-1, the proprotein convertase furin was coexpressed with proBACE-1, leading to almost complete proteolytic activation of the recombinant enzyme . The mature enzyme was secreted in the conditioned medium of BACE-1/furin coinfected HighFive insect cells . Secreted BACE-1 protein was purified to homogeneity from the medium using subsequent Ni-chelate affinity chromatography, anion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration . To avoid autoproteolysis, all purification steps were performed at pH values outside the activity range of BACE-1 . The purified, biologically active enzyme was homogeneous on SDS/PAGE and had the expected sequence and molecular mass determined by N-terminal amino acid sequencing and mass spectrometry, respectively . Moreover, the preparation showed a single peak of the expected size with only 17% polydispersity using dynamic light scattering analysis . The yield of BACE-1 from fermentation cultures was approximately 0.1mg pure enzyme per liter of cell culture medium . The purified protein was successfully used to generate BACE-1/inhibitor co-crystals and to determine the crystal structure of the complex by X-ray analysis . The availability of substantial quantities of active, homogeneous enzyme will be of great help in future structure-based drug design efforts in the search for efficient protease inhibitor drugs to treat AD.

Protein Expr Purif, 2002 Oct, 26(1), 14 - 8
An economical 20 litre bench-top fermenter; Thiel MA et al.; We describe an economical 20 litre bench-top fermenter suitable for production of recombinant antibody fragments in bacterial expression systems . The bacterial culture contained within a polycarbonate carboy is mixed (400-600 rpm) and aerated (1 vessel vol./min) by a high-shear radial flow impeller mounted on a hollow stainless steel shaft, through which pressurised air is pumped . Air is dispersed as fine bubbles into the culture medium by the turbine impeller, without the need for a porous sparger . A stainless steel baffle stabilised by a gliding counterweight increases mixing . The components can easily be disassembled for cleaning and sterilisation . Temperature (range 20-37 degrees C) and pH (range 7.0-7.5) are controlled manually . Using the apparatus, it proved possible to achieve Escherichia coli cell culture densities equivalent to an optical density at 600 nm (OD(600)) of 30-32, compared with OD(600) 4-6 in shake flasks . A yield of 40 mg/litre/day of a recombinant antibody fragment was obtained with the fermenter, which was 15-fold more than the yield of 2.5mg/litre/day achieved in shake flasks . The fermenter may be particularly suited for research purposes.

Biotechnol Bioeng, 2002 Dec 5, 80(5), 594 - 7
Noninvasive measurement of dissolved oxygen in shake flasks; Tolosa L et al.; Shake flasks are ubiquitous in cell culture and fermentation . However, conventional devices for measuring oxygen concentrations are impractical in these systems . Thus, there is no definitive information on the oxygen supply of growing cells . Here we report the noninvasive, nonintrusive monitoring of dissolved oxygen (DO) in shake flasks using a low-cost optical sensor . The oxygen-sensitive element is a thin, luminescent patch affixed to the inside bottom of the flask . The sensitivity and accuracy of this device is maximal up to 60% DO, within the range that is critical to cell culture applications . By measuring actual oxygen levels every 1 or 5 min throughout the course of yeast and E . coli fermentations, we found that a modest increase in shaker speed and a decrease in culture volume slowed the onset of oxygen limitation and reduced its duration . This is the first time that in situ oxygen limitation is reported in shake flasks . The same data is unattainable with a Clark type electrode because the presence of the intrusive probe itself changes the actual conditions . Available fiber optic oxygen sensors require cumbersome external connections and recalibration when autoclaved .

Biotechnol Bioeng, 2002 Dec 5, 80(5), 589 - 93
Production of lovastatin examined by an integrated approach based on chemometrics and DOSY-NMR; Bradamante S et al.; Microbial secondary metabolites are one of the sources of therapeutic molecules in the pharmaceutical industry . Product quality and high yields of secondary metabolites are the main goals for the commercial success of a fermentation process . Our novel approach was based on the decision-tree algorithm to determine the key variables correlated with the process outcome and on DOSY-NMR to identify both co-metabolites and impurities, and it improves fermentation systems and speeds up bioprocess development . The approach has been validated in the case of lovastatin production from Aspergillus terreus .

Biotechnol Bioeng, 2002 Dec 5, 80(5), 559 - 68
Continuous annular chromatography: General characterization and application for the isolation of recombinant protein drugs; Vogel JH et al.; Isolation of recombinant protein drugs from cell culture supernatant is usually performed in batch mode, even if the fermentation process itself is continuous . As a novel approach, continuous separation techniques like continuous annular chromatography (CAC) can be used for continuous isolation . The potential of CAC for industrial application is demonstrated by continuous isolation of rFVIII from cell culture supernatant in pilot scale (i.e., 144-288 l/day) . Thirty-fold concentration can be achieved at 94% yield, while purity is increased 3-5-fold . For this a batch direct feed ion exchange chromatography method was adapted to a commercial preparative CAC system (P-CAC) . A headspace loading technique was used to maximize the concentration factor, while buffer incompatibility problems were addressed by a specifically modified inlet geometry . To allow sterile on-line coupling to FVIII-producing perfusion fermenters, an autoclavable pilot scale P-CAC prototype was developed . General characterization of P-CAC revealed a current limitation of the technology, i.e., variations in the outlet flow rates of up to +/-20% . These flow variations are shown to be caused mainly by a nonuniform annular resin bed and in turn result in "peak wobbling," i.e., the slight variation of peak position (up to +/-4 degrees ) and shape (e.g., A(s) = 0.9-1.4) as a specific function of column position . Some additional peak broadening, although less significant, is caused by a "peak oscillation" effect that results from the necessary segmentation of flow into discrete outlets . Both effects are only measurable if peaks are either monitored continuously or at least measured at multiple column positions . For isolation processes, these nonideal flow phenomena mean that more outlet streams have to be collected in order to achieve maximum yield and thus the achievable concentration factor is somewhat lower than the theoretical maximum .

Biotechnol Bioeng, 2002 Dec 5, 80(5), 525 - 36
Modeling acidogenic and sulfate-reducing processes for the determination of fermentable fractions in wastewater; Ruel SM et al.; The biochemical acidogenic potential (BAP) of a wastewater is the maximum concentration of volatile fatty acids (VFAs) that can be measured at the end of an anaerobic fermentation test . A model was constructed to describe the acidogenic reactions occurring during BAP tests and to divide the BAP into organic fractions . The model was calibrated with a set of specific experiments highlighting the role of sulfate-reducing bacteria on acidogenic processes, which description was necessary for correct parameter identification . The model could describe acidogenic fermentation processes, with or without sulfate reduction, at 20 degrees C, for 13 wastewaters of different origin, composition, and settleability using the same optimized parameters . A simplified version of the model, without sulfate reduction, was able to describe VFA production by the adjustment of only three variables: readily fermentable organic matter (Sf), anaerobically hydrolyzable organic matter (Xf), and heterotrophic acidogenic biomass (Xha), which proved to be coherent with the experimental BAP value . The combination of the BAP test and the model developed in this study resulted in a new reliable tool to characterize wastewater under anaerobic conditions . As VFAs are the main substrates for phosphate-accumulating organisms (PAOs), the use of organic fractions VFA, Sf, Xf, and Xha in wastewater treatment plant modeling could improve the predictability and optimization of enhanced biological phosphorus removal (EBPR) processes .

J Ind Microbiol Biotechnol, 2002 Oct, 29(4), 189 - 95
Factors affecting the yield and properties of bacterial cellulose; Krystynowicz A et al.; Acetobacter xylinum E(25) has been applied in our studies in order to find optimal culture conditions for effective bacterial cellulose (BC) production . The strain displays significantly higher stability in BC production under stationary culture conditions . In contrast, intensive agitation and aeration appear to drastically reduce cellulose synthesis since such conditions induced formation of spontaneous cellulose nonproducing mutants (Cel-), which dominated in the culture . Mutation frequency strictly depends on the medium composition in agitated cultures . Enrichment of the standard SH and Yamanaka media with 1% ethanol significantly enhanced BC production in stationary cultures . Horizontal fermentors equipped with rotating discs or rollers were successfully applied in order to improve culture conditions . Relatively slow rotation velocity (4 rpm) and large surface area enabling effective cell attachment are optimal parameters for cellulose production . Physical properties of BC samples synthesized either in stationary cultures or in a horizontal fermentor revealed that cellulose from stationary cultures demonstrated a much higher value of Young's modulus, but a much lower value of water-holding capacity.

J Ind Microbiol Biotechnol, 2002 Oct, 29(4), 181 - 4
Optimization of exopolysaccharide production by Tremella mesenterica NRRL Y-6158 through implementation of fed-batch fermentation; De Baets S et al.; In liquid culture conditions, the yeast-like fungus Tremella mesenterica occurs in the yeast state and synthesizes an exopolysaccharide (EPS) capsule, which is eventually released into the culture fluid . It is composed of an alpha-1,3-D-mannan backbone, to which beta-1,2 side chains are attached, consisting of D-xylose and D-glucuronic acid . Potato dextrose broth (PDB) seemed to be an excellent medium for both growth of the yeast cells and synthesis of the EPS . This medium is composed solely of an extract of potatoes to which glucose was added . Yet an important disadvantage of this production medium is the presence of starch in the potato extract, since Tremella cells are not capable of metabolizing this component; furthermore, it coprecipitates upon isolation of the polymer {3} . In this respect, it was essential to remove the starch in order to achieve high polysaccharide production and recovery . A good method was the removal of starch through ultrafiltration of the PDB medium before inoculation of the strain . This resulted in an excellent starch-free medium in which other components essential for polysaccharide production were still present {3} . Through implementation of single and cyclic fed-batch fermentations with glucose feed, 1.6- and 2.2-fold increases in EPS yield were obtained, respectively . Lowering the carbon source level by using a cyclic fed-batch technique might decrease the osmotic effect of glucose or any catabolite regulation possibly exerted by this sugar on enzymes involved in EPS synthesis.

J Ind Microbiol Biotechnol, 2002 Oct, 29(4), 149 - 54
A review of antifoam mechanisms in fermentation; Pelton R; Antifoam (defoamer) chemicals are a crucial part of many commercial fermentation processes . Reviewed are the types of defoamers and their mode of operation . Also presented is a simple model, which simulates foam growth as functions of defoamer concentration, air hold-up, reactor volume and air flow rate.

Syst Appl Microbiol, 2002 Aug, 25(2), 287 - 93
Analysis of yeast populations during alcoholic fermentation: a six year follow-up study; Beltran G et al.; Wine yeasts were isolated from fermenting Garnatxa and Xarel.lo musts fermented in a newly built and operated winery between 1995 and 2000 . The species of non-Saccharomyces yeasts and the Saccharomyces cerevisiae strains were identified by ribosomal DNA and mitochondrial DNA RFLP analysis respectively . Non-Saccharomyces yeasts, particularly Hanseniaspora uvarum and Candida stellata, dominated the first stages of fermentation . However Saccharomyces cerevisiae was present at the beginning of the fermentation and was the main yeast in the musts in one vintage (1999) . In all the cases, S . cerevisiae took over the process in the middle and final stages of fermentation . The analysis of the S . cerevisiae strains showed that indigenous strains competed with commercial strains inoculated in other fermentation tanks of the cellar . The continuous use of commercial yeasts reduced the diversity and importance of the indigenous S . cerevisiae strains.

Biosci Biotechnol Biochem, 2002 Aug, 66(8), 1697 - 705
New convulsive compounds, brasiliamides A and B, from Penicillium brasilianum batista JV-379; Fujita T et al.; New convulsive compounds, brasiliamides A (1) and B (2), were isolated by activity-guided fractionation from okara fermented with a soil isolate of Penicillium brasilianum Batista JV-379 . Their structures were elucidated on the basis of spectral and chemical evidence and by X-ray crystallography of the hydrogenated product of 2 . In the 1H- and 13C-NMR spectra of 2, the signals were complicated, all being doubled or broadened in several deuterated solvents at room temperature . The conformational change of 2 was clarified as the rotational isomerization of amide bonds in solution by NMR measurements at various temperatures . Four rotamers of 2 at two amide bonds were presented at -60 degrees C in CDCl3, whereas only two isomers were apparent at room temperature, owing to rapid rotation of one of the amide bonds . Brasiliamides A and B respectively showed convulsive activity against silkworms with ED50 values of 300 and 50 microg/g of diet.

J Pediatr Gastroenterol Nutr, 2002 Sep, 35(3), 324 - 8
Quantitation of colonic luminal synthesis of butyric acid in piglets; Kien CL et al.; OBJECTIVES: Butyric acid, synthesized via bacterial fermentation in colonic lumen, may play an important role in the nutrition of the colonic mucosa . Since disaccharides, especially lactose, are the principal dietary carbohydrates during infancy, it is important to determine if their fermentation is associated with butyric acid synthesis . The objective of this paper is to describe a newly developed stable isotope method for quantifying butyric acid synthesis in the colonic lumen and to demonstrate its application during cecal infusions of lactose and lactulose in piglets . METHODS: Nine piglets aged 21 to 30 days were studied during acute anesthesia . The C enrichment of butyric acid was monitored in the portal vein before and during a 120 minutes cecal infusion of {1- C}-butyric acid and either unlabeled lactose (N = 4) or lactulose (N = 5) . RESULTS: The luminal synthesis of BA (micromol x kg x min ) (Mean +/- S.D.) was respectively 1.5 +/- 0.9 and 1.2 +/- 0.6 during lactulose and lactose infusion . CONCLUSIONS: This study provides new quantitative data on in vivo butyric acid production in the mammalian colon.

J Biol Chem, 2002 Nov 29, 277(48), 46408 - 14 Epub 2002 Sep 25.
Fermented wheat germ extract inhibits glycolysis/pentose cycle enzymes and induces apoptosis through poly(ADP-ribose) polymerase activation in Jurkat T-cell leukemia tumor cells; Comin-Anduix B et al.; The fermented extract of wheat germ, trade name Avemar, is a complex mixture of biologically active molecules with potent anti-metastatic activities in various human malignancies . Here we report the effect of Avemar on Jurkat leukemia cell viability, proliferation, cell cycle distribution, apoptosis, and the activity of key glycolytic/pentose cycle enzymes that control carbon flow for nucleic acid synthesis . The cytotoxic IC(50) concentration of Avemar for Jurkat tumor cells is 0.2 mg/ml, and increasing doses of the crude powder inhibit Jurkat cell proliferation in a dose-dependent fashion . At concentrations higher than 0.2 mg/ml, Avemar inhibits cell growth by more than 50% (72 h of incubation), which is preceded by the appearance of a sub-G(1) peak on flow histograms at 48 h . Laser scanning cytometry of propidium iodide- and annexin V-stained cells indicated that the growth-inhibiting effect of Avemar was consistent with a strong induction of apoptosis . Inhibition by benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone of apoptosis but increased proteolysis of poly(ADP-ribose) indicate caspases mediate the cellular effects of Avemar . Activities of glucose-6-phosphate dehydrogenase and transketolase were inhibited in a dose-dependent fashion, which correlated with decreased (13)C incorporation and pentose cycle substrate flow into RNA ribose . This decrease in pentose cycle enzyme activities and carbon flow toward nucleic acid precursor synthesis provide the mechanistic understanding of the cell growth-controlling and apoptosis-inducing effects of fermented wheat germ . Avemar exhibits about a 50-fold higher IC(50) (10.02 mg/ml) for peripheral blood lymphocytes to induce a biological response, which provides the broad therapeutic window for this supplemental cancer treatment modality with no toxic effects.

FEMS Microbiol Lett, 2002 Sep 10, 214(2), 223 - 7
Effect of Vitreoscilla hemoglobin biosynthesis in Escherichia coli on production of poly(beta-hydroxybutyrate) and fermentative parameters; Yu H et al.; In order to attain high cell density and low cost production of poly(beta-hydroxybutyrate) (PHB), the Vitreoscilla globin gene (vgb) was introduced into a novel recombinant strain, Escherichia coli VG1 (pTU14) . Experiments showed that the expression of vgb was under the regulation of dissolved oxygen (DO) in broth and the introduction of vgb in VG1 (pTU14) induced the parent promotion effect on cell growth and PHB accumulation, especially under low DO conditions . Further experiments indicated that the introduction of vgb in VG1 (pTU14) not only decreased the critical oxygen concentration, but also affected the volumetric oxygen transfer coefficient of the recombinant strain.

J Nat Prod, 2002 Sep, 65(9), 1274 - 7
Three novel cytochalasins X, Y, and Z from Pseudeurotium zonatum; Feng Y et al.; Fermentation of Pseudeurotium zonatum led to the isolation of the known cytochalasin G (1) and three new cytochalasins, X, Y, and Z (2-4) . These four compounds are the only naturally occurring cytochalasins reported to date that contain an indole-substituted perhydroisoindol-1-one fused with an 11-membered macrocycle.

J Anim Sci, 2002 Sep, 80(9), 2214 - 22
An in vitro study of manure composition on the biochemical origins, composition, and accumulation of odorous compounds in cattle feedlots; Miller DN et al.; Very little is known about the biochemical origin of cattle feedlot odors and the environmental factors controlling their production . The tie between diet and manure composition is well established, but the effect of different manure compositions on odorous chemical production is unknown . This study describes the effect of starch, casein, and cellulose substrate additions to slurries of fresh (< 24 h) and aged cattle manure (> 1 d) on the anaerobic production of fermentation products and the consumption of substrates relative to no addition treatments . Aged cattle manure accumulated more VFA (245 to 290 mM) than the fresh manure (91 to 181 mM) irrespective of substrate additions (P < 0.001) . In fresh manures, VFA concentrations were increased (P < 0.01) over no addition treatments when carbohydrate (starch or cellulose) was added, whereas starch and protein treatments to aged manure increased VFA content relative to no addition treatments (P < 0.001) . Branched-chain VFA and aromatic compounds accumulated only in the aged manure (no addition and protein treatments), indicating that some protein fermentation occurred in those treatments . Based upon substrate loss, starch fermentation was the dominant process in both manures and all treatments with losses exceeding 18.6 g/L . Protein fermentation occurred only in the aged manure, specifically the no addition and protein treatments, when starch was no longer available . The production of odorous compounds from manure was controlled by substrate availability and pH, with pH related to lactate accumulation . We believe that calcareous soil and lactate-consuming microorganisms in the aged manure slurries minimized slurry acidification and resulted in greater accumulations of odorous products . Substrate additions had little effect on the overall accumulation of odor compounds in manure but had profound effects on odor compound composition . We propose that modifying cattle diets to limit starch and protein excretion would profoundly affect the production and accumulation of odor compounds in feedlots.

Niger J Paediatr, 1993 Jan, 20(1), 1 - 5
Clinical trial of pap-salt solution in the treatment of dehydrated children; Olanrewaju DM et al.; PIP: In a randomized clinical trial the efficacy of Pap-salt solution (PSS) was compared with the World Health Organization (WHO) oral rehydration salt (ORS) solution in 212 children 6 months to 5 years old: 101 cases (52 males and 49 females) and 111 controls (41 males and 70 females) at the primary health care centers of Ijebu-Igbo, Ita-Oba in Sagamu, and at the oral rehydration therapy (ORT) unit of the Ogun State University Teaching Hospital, Sagamu, Nigeria, between June 1991 and June 1992 . 164 children (77.3%) belonged to the lowest socioeconomic classes . Maize pap (Ogi) was prepared by soaking maize in water for 2-3 days for fermentation to occur . PSS was prepared from raw pap and the cooked pap was then made up to 600 ml with the addition of more water and 3 ml of table salt . Each child received 75 ml per kg of ORS solution or PSS orally over a 4-hour period . Those patients who still showed signs of dehydration were given a further 75 ml/kg of the same solution over another 4-hour period . The patients were followed up on 2 consecutive days at home by community health nurses . All the 101 children who received PSS were successfully treated and the remaining 2 were lost to follow-up . There was no significant difference in the mean volumes of PSS (1015.4 ml) and ORS (1036 ml) required for rehydration (p 0.05) . 96 mothers found PSS easy to prepare and readily accepted by their children, and 97 stated that the ingredients required for preparation were available . There was no difference ( p 0.05) in the pre- and post-treatment values of the serum electrolytes in the 2 groups . PSS was acceptable to the patients, readily available, and much cheaper than the ORS solution . Therefore, PSS is recommended for the rehydration of children with mild dehydration in situations where ORS solution is not available .

Draper Fund Rep, 1983 Aug, (12), 16 - 8
Latin American perspectives on the individual and the greater community; Siqueira Wiarda I; PIP: Ethical questions have been at the heart of the ongoing debate over population policies and programs since the inception of organized family planning services in Latin America in the early 1960s . An ethically acceptable balance needs to be found between the well-being of and respect for the individual and the welfare of the family within the larger society . In Latin America the family is the most enduring and respected institution . All indications are that the family will continue to dominate the life of Latin Americans long after the societies become modernized . The extended family has prevailed because it has met the needs of individuals in the group and in the community . The place of the individual in the extended kin network and in this type of society is determined by several conditions, not the least of which is the ability to adjust personal needs to the needs of the extended family . Women, especially, if they are married, are expected to bear children and thus reinforce and perpetuate family ties . Despite considerable talk and some action giving men equal responsibility for raising and supporting children, traditionally this is considered the major prerogative and obligation of Latin women . Latin women's movements, particularly in Brazil, Mexico, and Venezuela, have been at the forefront in redefining women's rights and responsibilities . This is especially the case in the area of reproductive rights . Such redefinitions, involving a new perspective on ethical values, will not be soon or easily accepted . Instrumental in ending traditions and discarding ethical orientations is the fast pace of social change . Out of the social ferment and rapid modernization and the questioning of traditions, new definitions of ethical issues concerning reproductive roles and responsibilities toward family and community are slowly emerging . The definitions and their practical meanings cluster in 4 main groups: the freedom to make reproductive choices; individual and community betterment; social justice; and individual dignity . In all these questions, there are legitimate rationales for childbearing or for contraceptive use from the individual's perspective, but from the point of view of the global community, continued high fertility appears to be a longterm plan for disaster . By taking into account the greater good of the community, the exercise of individual choices takes on new meaning .

Biotechnol Bioeng, 2002 Nov 20, 80(4), 419 - 27
Multiblock PLS analysis of an industrial pharmaceutical process; Lopes JA et al.; The performance of an industrial pharmaceutical process (production of an active pharmaceutical ingredient by fermentation, API) was modeled by multiblock partial least squares (MBPLS) . The most important process stages are inoculum production and API production fermentation . Thirty batches (runs) were produced according to an experimental planning . Rather than merging all these data into a single block of independent variables (as in ordinary PLS), four data blocks were used separately (manipulated and quality variables for each process stage) . With the multiblock approach it was possible to calculate weights and scores for each independent block . It was found that the inoculum quality variables were highly correlated with API production for nominal fermentations . For the nonnominal fermentations, the manipulations of the fermentation stage explained the amount of API obtained (especially the pH and biomass concentration) . Based on the above process analysis it was possible to select a smaller set of variables with which a new model was built . The amount of variance predicted of the final API concentration (cross-validation) for this model was 82.4% . The advantage of the multiblock model over the standard PLS model is that the contributions of the two main process stages to the API volumetric productivity were determined .

Appl Environ Microbiol, 2002 Oct, 68(10), 5186 - 90
Acetate utilization and butyryl coenzyme A (CoA):acetate-CoA transferase in butyrate-producing bacteria from the human large intestine; Duncan SH et al.; Seven strains of Roseburia sp., Faecalibacterium prausnitzii, and Coprococcus sp . from the human gut that produce high levels of butyric acid in vitro were studied with respect to key butyrate pathway enzymes and fermentation patterns . Strains of Roseburia sp . and F . prausnitzii possessed butyryl coenzyme A (CoA):acetate-CoA transferase and acetate kinase activities, but butyrate kinase activity was not detectable either in growing or in stationary-phase cultures . Although unable to use acetate as a sole source of energy, these strains showed net utilization of acetate during growth on glucose . In contrast, Coprococcus sp . strain L2-50 is a net producer of acetate and possessed detectable butyrate kinase, acetate kinase, and butyryl-CoA:acetate-CoA transferase activities . These results demonstrate that different functionally distinct groups of butyrate-producing bacteria are present in the human large intestine.

Appl Environ Microbiol, 2002 Oct, 68(10), 5136 - 41
Direct and efficient production of ethanol from cellulosic material with a yeast strain displaying cellulolytic enzymes; Fujita Y et al.; For direct and efficient ethanol production from cellulosic materials, we constructed a novel cellulose-degrading yeast strain by genetically codisplaying two cellulolytic enzymes on the cell surface of Saccharomyces cerevisiae . By using a cell surface engineering system based on alpha-agglutinin, endoglucanase II (EGII) from the filamentous fungus Trichoderma reesei QM9414 was displayed on the cell surface as a fusion protein containing an RGSHis6 (Arg-Gly-Ser-His(6)) peptide tag in the N-terminal region . EGII activity was detected in the cell pellet fraction but not in the culture supernatant . Localization of the RGSHis6-EGII-alpha-agglutinin fusion protein on the cell surface was confirmed by immunofluorescence microscopy . The yeast strain displaying EGII showed significantly elevated hydrolytic activity toward barley beta-glucan, a linear polysaccharide composed of an average of 1,200 glucose residues . In a further step, EGII and beta-glucosidase 1 from Aspergillus aculeatus No . F-50 were codisplayed on the cell surface . The resulting yeast cells could grow in synthetic medium containing beta-glucan as the sole carbon source and could directly ferment 45 g of beta-glucan per liter to produce 16.5 g of ethanol per liter within about 50 h . The yield in terms of grams of ethanol produced per gram of carbohydrate utilized was 0.48 g/g, which corresponds to 93.3% of the theoretical yield . This result indicates that efficient simultaneous saccharification and fermentation of cellulose to ethanol are carried out by a recombinant yeast cells displaying cellulolytic enzymes.

Appl Environ Microbiol, 2002 Oct, 68(10), 5034 - 41
Improvement of recombinant protein yield by a combination of transcriptional amplification and stabilization of gene expression; Cebolla A et al.; We explored the use of a cascade circuit for heterologous gene expression that consists of a regulatory module with a salicylate-inducible system that controls the expression of a second regulator, xylS2, whose product is activated by common inducers . Activation and increasing the concentration of the second regulator synergistically induced heterologous genes downstream of the Pm promoter in the expression module . This module can be placed in multicopy vectors or in the chromosome of a host strain by means of minitransposons . Using reporter genes, we evaluated gene regulation capacity and gross production of the system with different configurations . The highest yield was obtained when the expression module was in a multicopy plasmid after a 6-h induction . However, expression modules in plasmids showed low stability after induction even with selective pressure . The chromosomal configuration had the lowest basal levels and induced levels comparable to those of plasmid configurations, resulting in accumulation of more than 10% of the total protein . Unlike the configurations in plasmids, the yield was maintained for at least 3 days even without selective pressure . In conclusion, the cascade system in the chromosome configuration is more efficient for long-term fermentation because of the great stability of the overexpressing phenotype in spite of the high levels of expression.

Appl Environ Microbiol, 2002 Oct, 68(10), 5012 - 6
Differential inhibition of mollicute growth: an approach to development of selective media for specific mollicutes; Keceli SA et al.; The energy-generating pathways of Mycoplasma spp . are diverse . Thus, it was predicted that the ability of inhibitors of these pathways to block growth would vary among species . This prediction was tested with 14 Mycoplasma species and potential inhibitors . The greatest differentiation among test species was obtained using fluoride, iodoacetate (IAA), beta-fluoropyruvate (FP), cibacron blue (CB), L-citrulline, and carbonyl cyanide m-chlorophenylhydrazone . A range of other potential inhibitors, including L-arginine analogues, had little inhibitory effect on growth, and D-arginine was shown to be a growth substrate for arginine-hydrolyzing species . Fluoride selectively inhibited the growth of mycoplasmas that were able only to ferment sugars to lactate and/or to hydrolyze arginine . In contrast, IAA was most effective against organic acid-oxidizing species, and L-citrulline inhibited arginine-hydrolyzing species . Mycoplasma verecundum, a species for which energy sources have not been identified, was relatively resistant to FP . Similarly, Acholeplasma laidlawii was distinguished by its CB resistance.

Ann Bot (Lond), 2002 Oct, 90(4), 499 - 507
Impact of oxygen stress and energy availability on membrane stability of plant cells; Rawyler A et al.; This article reviews the relationship between the energy status of plant cells under O(2) stress (e.g . waterlogging) and the maintenance of membrane intactness, using information largely derived from suspension cultures of anoxia-intolerant potato cells . Energy-related parameters measured were fermentation end-products (ethanol, lactate, alanine), respiratory rate, ATP, adenylate energy charge, nitrate reductase activity and biomass . ATP synthesis rates were calculated from the first four parameters . Reactive oxygen species were estimated from H(2)O(2) and superoxide levels, and the enzymatic detoxification potential from the activity levels of catalase and superoxide dismutase . Structure-related parameters were total fatty acids, free fatty acids (FFAs), lipid hydroperoxides, total phospholipids, N-acylphosphatidylethanolamine (NAPE) and cell viability . The following issues are addressed in this review: (1) what is the impact of anoxia on membrane lipids and how does this relate to energy status; (2) does O(2) per se play a role in these changes; (3) under which conditions and to what extent does lipid peroxidation occur upon re-aeration; and (4) can the effects of re-aeration be distinguished from those of anoxia? The emerging picture is a reappraisal of the relative contributions of anoxia and re-aeration . Two successive phases (pre-lytic and lytic) characterize potato cells under anoxia . They are connected by a threshold in ATP production rate, below which membrane lipids are hydrolysed to FFAs, and NAPE increases . Since lipid peroxidation occurs only when cells are reoxygenated during the lytic phase, its biological relevance in an already damaged system is questionable.

CVI Forum, 1996 Aug, (12), 10 - 3
Plant vaccines: edible, but how credible?
The effect of a nutrition education programme on pregnant women attending Khartoum model clinic.
PIP: A comparative study conducted in the Sudan illustrated both the potential of a nutrition education program for pregnant women and the many obstacles to creating more optimal maternal nutrition . Included in the study were 1200 women 18-34 years of age attending the Khartoum Province Model Clinic for a minimum of 5 prenatal visits . 83.3% were Muslims and 48.2% were illiterate . The 60 women who comprised the experimental group were exposed to individual nutrition counseling and home visits . Overall, 70% of subjects were anemic, 27% had malaria, and 18% were hookworm . The average weight gain during pregnancy was 4.5 kgs; 25.0% of controls and 20.0% of those in the experimental group gained 3.5-4.0 kgs, 18.3% of controls and 33.3% of experimental subjects gained 4.5-5.0 kgs, and 23.3% of controls and 30.0% of those in the experimental groups gained 5.5 kgs or more . 53.0% of controls and 61.7% of women exposed to nutrition education breastfed for at least 6 months, and 70% of women in the former group and 95% of those in the latter had enrolled their infants in an immunization program . A baseline nutrition questionnaire revealed widespread misinformation, especially a belief that pregnant women should reduce their food intake from 3 to 1-2 meals per day to prevent the fetus from growing too large and making labor difficult . Foods such as fermented sorghum products, fresh fish, and water melon were regarded as abortifacients and avoided, while hot pepper, sour milk, and cola sodas were considered nutritious . Consumption was self-reported to be poor for milk products by 25% of mothers, for meat by 16%, and for fruits and vegetables by 40% Although the educational intervention increased women's knowledge of basic nutrition, it had only a minimal effect on actual practice . This finding is assumed to reflect the widespread belief that men and their guests should be given preference in food allocation, poverty, and avoidance, on the basis of tradition, of many healthy foods . Since 72% of women reported ownership of a television, use if this medium is recommended for nutrition education .

Chin Sociol Anthropol, 1984 Spring-Summer, 16(3-4), 1 - 42
Demography, population growth, and population policy in China: a brief history from 1940 to the present; Chu DS; PIP: For some 3 decades after the founding of the People's Republic of China, Chinese studies have had to life with the paradoxical situation in which the abundance of population on the mainland, ever increasing in number predictably and inexorably, was couples with a scarcity of data about the nature and patterns of demographic change, ever harder to come by . As increasing amounts of new demographic data from China, sometimes produced jointly between Chinese and foreign researchers, become available, it is clear that previous methods, technics, and styles of analyses habituated by 3 decades of minimal data from China will become inadequate and obsolete . In 1957 Ma Yinchu put forth 3 main proposals for population control: 1) to conduct another national census to gain more demographic statistics, 2) to advocate widespread public education to inform people of the importance and practical knowledge of birth control combined with propaganda on the disadvantages of early marriages and the benefits of late marriage, and 3) family planning, by popular use of contraception, is the most effective method for population control . It is argued that 1) the years 1950-1958 consisted of a period of fermentation and preparation, 2) the post-leap years, 1959-1965, constituted a period of trial implementation of emerging policies, 3) 1966-1971 was an uncontrolled period, and 4) only the years after 1971 may be viewed as a period during which serious implementation took place . Overall, the selections in this volume are intended to inform both China scholars and social scientists interested in recent developments in Chinese population studies .

SCN News, 1997 Dec, (15), 25 - 6
Tulimbe Nutrition Project: a community-based dietary intervention to combat micronutrient malnutrition in rural southern Malawi; Berhe G; PIP: This article describes the community-based nutrition intervention in rural southern Malawi . The program aims to reverse micronutrient deficiencies in vitamin A, iron, and zinc in a society where staple diets are plant-based and contain high levels of anti-nutrients . Anti-nutrients, such as polyphenols, dietary fiber, and phytates, inhibit absorption of iron and zinc . This population's diet was also low in dairy and meat products . The Tulimbe Nutrition Project aimed to modify and diversify diets rather than to supplement or fortify diets . This approach was more culturally acceptable and economically feasible . The approach required changing food selection patterns and methods of preparing and processing indigenous foods . The new diets aimed to enhance the availability, access, and use of micronutrient-rich foods throughout the year . The project was initiated in 1995 in two communities among 300 families with children ranging in age from 3 to 7 years . A baseline assessment with interviews and focus groups was conducted . The assessment for children included a 24-hour dietary recall, anthropometric measurement, and other clinical measurement . Anthropometric and dietary assessments were repeated at 6 and 12 months . New cultivars and technologies were introduced, such as soybeans, short-duration pigeon peas, groundnuts, sunflower seeds, and papaya seedlings . The Malawi Industrial Research and Technology Development Center built and installed solar dryers, seed oil presses, and ovens in each community . People were encouraged to include soaked and fermented maize flour and germinated cereal flours in infant and child porridges . Parents were educated about micronutrient-rich foods, meal frequencies, portion sizes, and food combinations . Information was provided through demonstrations, home visits, plays, songs, and booklets . The program evaluation is in progress .

AIDS Action, 1995 Sep-Nov, (30), 4 - 5
Eat healthily, stay healthy; Making weaning safer; PIP: Infants are at increased risk of infection during weaning when caregivers begin giving foods other than breast milk to 4-6 month old infants . Since the gradual withdrawal of breast milk reduces its benefits, infants are more exposed to food-borne microorganisms . Thus, it is especially important to practice safe food preparation and storage during weaning . An infant's stomach requires caregivers to feed infants 5-6 small meals/day . Many mothers, however, cannot cook fresh food 5-6 times/day due to an already heavy workload . Insufficient time and scarce or costly fuel requires many families to cook large amounts of food and store it for subsequent meals . They should use safe storage methods . They should cover foods to protect the food from contamination by flies or dust . To keep microorganisms from proliferating, the food should be kept cool, but most families do not have a refrigerator . There are traditional ways to preserve and protect food, however . Storing foods in running water or in porous pots standing in water keep the food cool . Storing food in baskets hanging from the ceiling keep insects and rodents away . It also allows the wind to cool it . The traditional preparation methods of fermentation and production of curd from milk provide an environment unfavorable to the growth of pathogenic bacteria . Heating and reheating food fully destroys most pathogens that cause diarrhea . Improved cookstoves allow families on a limited budget to thoroughly heat food before serving it . Health workers and educators should familiarize themselves with local traditional food practices, particularly those that contribute to food safety . They should also work with communities to find ways to reduce the risk of diarrhea from contaminated food .

Nutr Rep Int, 1986 Nov, 34(5), 915 - 9
Proposed weaning formula for developing countries; Hafez YS et al.; PIP: This article proposes a process of combining soybean, corn and wheat to make an inexpensive weaning formula, and suggests that animal feeding studies, human organoleptic (taste) tests and finally feeding trials on infants be conducted . The method combines 3 steps involving soybean yogurt, corn starch solution and cracked wheat gruel . First a yogurt is fermented from cracked soybeans cooked in sodium bicarbonate solution, with sucrose added, and homogenized . Then corn is cracked, soaked, boiled, homogenized and sieved . Wheat bulgar (cooked cracked wheat) is powdered and boiled . The soy, corn and wheat components are mixed 3:2:1, sterilized and packaged . These operations would require equipment to mill, cook, time, measure and adjust pH, incubate, homogenize, sieve, weigh, compute dry weight, sterilize and package the formula . The authors tabulate published amino acid analysis of soy, corn and wheat . They propose to test the formula on laboratory animals to determine the feed efficiency ratio compared to standard chow diets, to test organoleptic quality on adult humans and finally to test the formula on weaned infants . It is anticipated that this formula will be inexpensive and appropriate for children with lactose or galactose intolerance or milk allergy .

Contracept Fertil Sex (Paris), 1975 June-July, 3(4), 271 - 3
{Demand for abortion and responses between the vote of the French law and its application}; Geraud R; PIP: The abortion law has not been passed yet but there is a general fermentation and legal pressure around it . There are 2 main contradictory opinions: some people think that the patient should prescribe herself her own therapeutics and the medical staff has the obligation to execute them, which means and antimedical solution . Others think that the medical staff can prescribe and execute . Both attitudes are unacceptable; the decision should be based on objective facts . The demand for aboriton has increased considerably in the last 2 years largely due to improper information on contraceptives . Television should be used for the dissemination and education on contraception . Too much has been written about abortion by politicians, theologians, philosophers, and journalists, while the medical specialists can hardly publish an objective article on aboriton in a reputable magazine . However, while controversy is stimulating for the medical staff, it is the patients who suffer the consequences . Therefore a solution of the problem is mandatory . The people should be informed and educated before a logical solution can be adopted .

World Environ Rep . 1975 Sep 1;1(16):6.
Haiti is an object lesson in ecological disaster; Candell A; PIP: The history of Haiti is summarized with emphasis on the slave ferment which destroyed every vestage of the French in 1804 and the slow ecological decline . The forests are now depleted . Haitians use charcoal for cooking and have denuded the moutains, no attempts have been made to check the resulting erosion, and today the 5 million people are among the poorest on earth . Haiti exported more coffee in 1914 that in 1973, sisal exparts dropped from 33.4 million kilos in 1950 to 4.1 million in 1972 and in 1974 it had to import rice to feed its people . The country cannot cope with the massive ecological disaster and not enough foreign aid is forthcoming .

Planta, 2002 Sep, 215(5), 754 - 62 Epub 2002 Jul 27.
Molecular and biochemical characterisation of two aspartic proteinases TcAP1 and TcAP2 from Theobroma cacao seeds; Laloi M et al.; Aspartic proteinase (EC 3.4.23) activity plays a pivotal role in the degradation of Theobroma cacao L . seed proteins during the fermentation step of cacao bean processing . Therefore, this enzyme is believed to be critical for the formation of the peptide and amino acid cocoa flavor precursors that occurs during fermentation . Using cDNA cloning and northern blot analysis, we show here that there are at least two distinct aspartic proteinase genes ( TcAP1 and TcAP2) expressed during cacao seed development . Both genes are expressed early during seed development and their mRNA levels decrease towards the end of seed maturation . TcAP2 is expressed at a much higher level than TcAP1, although the expression of TcAP1 increases slightly during germination . The proteins encoded by TcAP1 and TcAP2 are relatively different from each other (73% identity) . This, and the fact that the two corresponding genes have different expression patterns, suggests that the TcAP1 and TcAP2 proteins may have different functions in the maturing seeds and during germination . Because the TcAP2 gene is expressed at a much higher level during seed development than TcAP1, it is likely that the TcAP2 protein is primarily responsible for the majority of the industrially important protein hydrolysis that occurs during cacao bean fermentation . Finally, TcAP2 has been functionally expressed in the yeast Yarrowia lipolytica . The secreted recombinant protein is able to hydrolyse bovine haemoglobin at acidic pH and is sensitive to pepstatin A, confirming that TcAP2 encodes an aspartic proteinase, and strongly suggests that this gene encodes the well-characterized aspartic proteinase of mature cacao seeds.

J Biol Chem, 2002 Nov 22, 277(47), 44870 - 6 Epub 2002 Sep 19.
Bcl-x(L) complements Saccharomyces cerevisiae genes that facilitate the switch from glycolytic to oxidative metabolism; Vander Heiden MG et al.; All eukaryotic organisms have mechanisms to adapt to changing metabolic conditions . The mammalian cell survival gene Bcl-x(L) enables cells to adapt to changes in cellular metabolism . To identify genes whose function can be substituted by Bcl-x(L) in a unicellular eukaryote, a genetic screen was performed using the yeast Saccharomyces cerevisiae . S . cerevisiae grows by anaerobic glycolysis when glucose is available, switching to oxidative phosphorylation when carbohydrate in the media becomes limiting (diauxic shift) . Given that Bcl-x(L) appears to facilitate the switch from glycolytic to oxidative metabolism in mammalian cells, a library of yeast mutants was tested for the ability to efficiently undergo diauxic shift in the presence and absence of Bcl-x(L) . Several mutants were identified that have a defect in growth when switched from a fermentable to a nonfermentable carbon source that is corrected by the expression of Bcl-x(L) . These genes include the mitochondrial chaperonin TCM62, as well as previously uncharacterized genes . One of these uncharacterized genes, SVF1, promotes cell survival in mammalian cells in response to multiple apoptotic stimuli . The finding that TCM62 and the analogous human prohibitin gene also inhibit mammalian cell death following growth factor withdrawal implicates mitochondrial chaperones as regulators of apoptosis . Further characterization of the genes identified in this screen may enhance our understanding of Bcl-x(L) function in mammalian cells, and of cell survival pathways in general.

J Antibiot (Tokyo), 2002 Jul, 55(7), 629 - 34
A novel neuritogenic compound, NGA0187; Nozawa Y et al.; A new neuritogenic compound NGA0187 was isolated from the fermentation broth of Acremonium sp . TF-0356 . The structure of NGA0187 was determined by means of spectroscopic analysis and X-Ray diffraction . NGA0187 induced significant neurite outgrowth in PC12 cells . However, survival effect of NGA0187 on the primary culture of cerebral cortical neurons was not observed.

Biotechnol Appl Biochem, 2002 Oct, 36(Pt 2), 111 - 7
A new method for the preparation of human parathyroid hormone 1-34 peptides; Xiu Z et al.; An engineered Escherichia coli strain, BL21 (DE3)/pGEX-4T-human parathyroid hormone (hPTH) (1-34), was constructed by oligonucleotide annealing and PCR amplification of the target gene, and then by ligating it with the pGEX-4T-3 vector and transferring into the BL21 host . The soluble glutathione S-transferase (GST) fusion protein GST-hPTH (1-34), expressed from BL21 (DE3)/pGEX-4T-hPTH (1-34), was harvested after fermentation and purification by affinity chromatography . Following double cleavage by thrombin and prolyl endopeptidase, about 0.6 g/l intact hPTH (1-34) was harvested . The product was checked by HPLC MS and N-terminal sequence analysis . The purified recombinant hPTH (1-34) stimulates adenylate cyclase in rabbit renal cortical cell membranes to exactly the same extent as synthetic hPTH standards, indicating that the recombinant product has full biological activity.

Mol Biol Rep, 2002, 29(1-2), 203 - 9
Experimental supply-demand analysis of anaerobic yeast energy metabolism; Kroukamp O et al.; Experimental supply-demand analysis of yeast fermentative energy metabolism shows that control of the glycolytic flux is shared between supply and demand . In glucose limited chemostat cultures the supply block was modulated in a dilution rate change and demand block via a benzoic acid titration . Under these conditions the supply block had a flux control of 0.90 and the demand block a flux control of 0.10.

Z Naturforsch {C}, 2002 Jul-Aug, 57(7-8), 666 - 70
Evaluation of filamentous fungi and inducers for the production of endo-polygalacturonase by solid state fermentation; Dartora AB et al.; Aspergillus oryzae CCT 3940, Aspergillus awamori NRRL 3112 and a Trichoderma sp.) were compared for their capacity to produce endo-polygalacturonase (endo-PG) in solid state fermentation . Maximum pectinolytic activity was reached in 72 h of growth, the best two fungal strains being A . niger T0005007-2 and A . oryzae CCT 3940 . Three types of commercial purified pectin and four of unprocessed pectin (tangerine, orange, Tahiti lime and sweet lime rind) were used to assess the effect of pectin on the production of endo-PG by A . niger T0005007-2 . Maximum pectinolytic activity was achieved using 6 and 10% (w/w) of purified pectin as inducer . Depending on the origin of the commercial pectin used as inducer, maximum endo-PG levels varied from 223 to 876 units per gram of dry medium (one endo-PG unit (U) was defined as the quantity of enzyme which caused a reduction in viscosity of 50% in a 1% w/v solution of pectin in 30 min), indicating that care should be taken when choosing this component of the medium . When the crude pectins were used as inducers at the same concentration as purified pectin, maximum endo-PG activities were 250-300 units/g . However, by increasing the amount of Tahiti lime rind to 50% (w/w) maximum endo-PG was 919 U/g, thus opening up the possibility of a low cost medium for endo-PG production.

Chir Ital, 2002 Jul-Aug, 54(4), 555 - 8
Small bowel necrosis associated with postoperative enteral feeding; Zetti G et al.; Enteral feeding by jejunostomy is one of the main surgical procedures used to supply the proteins and calories necessary in the early postoperative period after major surgery of the upper digestive tract . The complications associated with early postoperative enteral feeding may vary from signs of gastrointestinal intolerance such as nausea, emesis, diarrhoea and cramp-like abdominal pain to hypotension and hypovolaemic shock, and also to the development of small bowel ischaemia and necrosis . Ischaemic intestinal involvement with progression towards necrosis is fortunately a rare event . The cause is not well known . A multifactorial pathogenesis of the mucosal damage has been proposed, where hyperosmolarity of feeding and bacterial overgrowth, due to excessive fermentation of carbohydrates, a decreased mesenteric blood flow and a lowering of peristalsis have been adduced as causes of mucosal injury . We report a case of intestinal necrosis following a jejunostomy procedure, which led to ileal resection.

J Agric Food Chem, 2002 Sep 25, 50(20), 5649 - 53
Effect of different strains of Saccharomyces cerevisiae on production of volatiles in Napa Gamay wine and Petite Sirah wine; Patel S et al.; Napa Gamay grapes were fermented with four different strains of the yeast Saccharomyces cerevisiae (VL1, MI16, Fermirouge, and RA17) . Petite Sirah grapes were fermented with seven different strains of the same yeast (BM45, Fermirouge, RA17, NI, CX3079, A350, and A796) . Volatile compounds formed in the wines were analyzed by gas chromatography/mass spectrometry . Volatile compounds found in both wines were alcohols, esters, and acids, as well as some miscellaneous compounds . Isoamyl alcohol was the compound found in the highest relative amount with all four yeast strains in the Napa Gamay wines, followed by 2-phenyl ethanol, monoethyl succinate, and hexanoic acid . The relative amounts of isoamyl alcohol ranged from 30.84% (VL1) to 43.28% (RA17) . Major volatile compounds found in Petite Sirah wines were isoamyl alcohol, 2-phenyl ethanol, 2-hydroxy ethyl propanoate, monoethyl succinate, and octanoic acid . The several esters, including 2-hydroxyethyl propanoate, may contribute to the fruity flavor of Petite Sirah wines . Overall, the S . cerevisiae yeast strains used to ferment Napa Gamay grapes and Petite Sirah grapes produced the same major components, with certain variations in formation levels.

J Agric Food Chem, 2002 Sep 25, 50(20), 5600 - 4
Foam aptitude of trepat and monastrell red varieties in cava elaboration . 2 . Second fermentation and aging; Girbau-Sola T et al.; The foam properties of sparkling wines (Cava) made from two red autochthonous grape varieties, Trepat and Monastrell, and coupages, including different percentages of them, were studied during second fermentation and aging . The effect of second fermentation on foam gave the highest decreases when the base wines had the highest foam values, while gave the lowest decreases or even increases for the base wines with the lowest foam characteristics . However, the greater the HM and Sigma of the base wine, the greater the foam values of the sparkling wine . Base wine determinations for quality control in cellars could provide information about future sparkling wine foaming . Acidity parameters, ethanol, sulfur, and polysaccharides contents were correlated to foam characteristics in the sparkling wines . In terms of color and foaming, wines made from the red varieties Trepat and Monastrell blended with white variety wines could be appropriate for elaborating "blanc de noirs" sparkling wines.

J Agric Food Chem, 2002 Sep 25, 50(20), 5596 - 9
Foam aptitude of trepat and monastrell red varieties in cava elaboration . 1 . Base wine characteristics; Girbau-Sola T et al.; The foam properties of base wines made from red autochthonous varieties (Trepat and Monastrell) were studied . Four wines of each variety were elaborated (fermented off skins at industrial scale in two consecutive harvests) and blended at different proportions with the white traditional variety (Macabeo, Xarel.lo, and Parellada) wines to elaborate Cava (closed-bottle-fermented sparkling wine) . When crescent amounts of Trepat were added to the traditional white blend, the foamability and the color intensity (CI) of the wine increased polinomically . The increase of the CI depended on the year of harvest . Thus, oenologists could decide the blend proportion most suitable to elaborate either a "blanc de noirs" sparkling wine or a new type of Cava.

J Chromatogr A, 2002 Aug 2, 965(1-2), 129 - 61
Preparative liquid chromatography; Guiochon G; The status of the theory and the main methods of implementation of preparative liquid chromatography are reviewed . On the theory front, the focus has recently shifted . The theory of non-linear, non-ideal chromatography has given rise to numerous models whose advantages, disadvantages and ranges of application are now well understood . Interest now resides in investigating the equilibrium thermodynamics of complex new systems, in the study of the kinetics of mass transfers in conventional chromatographic systems, and in the application of the various models of chromatography to optimize the experimental conditions . Progress in computer technology allows the use of sophisticated models, provided their parameters can be measured . This allows the detailed investigation of separations for which the mass transfer kinetics is slow such as chiral separations, the purification of basic compounds, and the extraction of recombinant proteins . On the applied front, in addition to numerous incremental improvements in reliability and economic performance, a few essential new features should be noted, i.e . the availability of instruments for simulated moving bed separations at the scale needed for preparative chiral separations, the use of expanded beds for the extraction of recombinant proteins from fermentation broths, and the attention given to improvements in the performance of packed beds . A survey of the literature dealing with practical applications and recent meetings shows that preparative chromatography is becoming a well established separation and purification method in the pharmaceutical industry.

Antimicrob Agents Chemother, 2002 Oct, 46(10), 3142 - 50
Mutations in 23S rRNA account for intrinsic resistance to macrolides in Mycoplasma hominis and Mycoplasma fermentans and for acquired resistance to macrolides in M . hominis; Pereyre S et al.; The mechanisms of intrinsic resistance of Mycoplasma hominis to 14- and 15-membered macrolides were investigated in comparison with those of M . pneumoniae, which is naturally susceptible to macrolides . Radiolabeled erythromycin was not accumulated by M . hominis PG21, but addition of an ABC transporter inhibitor increased the level of erythromycin uptake more than two times, suggesting the existence of an active efflux process . The affinity of {(14)C}erythromycin to ribosomes isolated from M . hominis was dramatically reduced relative to that to ribosomes isolated from M . pneumoniae . The nucleotide sequences of 23S rRNA of both ribosomal operons rrnA and rrnB and ribosomal proteins L4 and L22 of M . hominis were obtained . Compared to the sequence of M . pneumoniae, M . hominis harbored a G2057A transition in its 23S rRNA sequence, as did M . fermentans, another mycoplasma that is erythromycin resistant . An additional C2610U change was also found in the sequence of M . hominis . Moreover, two M . hominis clinical isolates with acquired resistance to 16-membered macrolides were examined for mutations in domain II and domain V of 23S rRNA and in ribosomal proteins L4 and L22 . Compared to the sequence of reference strain PG21, one isolate harbored a A2059G transition and a C2611U transition in one of the two rrn operons, while the other one was mutated only at position 2059, also on the same operon . No mutation was found in the two ribosomal protein sequences . Overall, the present study is an exhaustive characterization of the intrinsic resistance of M . hominis to 14- and 15-membered macrolides and the first description of mycoplasma clinical isolates resistant to macrolide, lincosamide, and streptogramin antibiotics harboring a mutation at position 2611 in the 23S rRNA.

Gene, 2002 Jul 10, 294(1-2), 13 - 24
Cloning and characterization of human and mouse SNRK sucrose non-fermenting protein (SNF-1)-related kinases; Kertesz N et al.; We previously isolated, from the earliest population of CD34+ hematopoietic progenitors that form in the aorta of the human embryo, a partial DNA complementary to RNA (cDNA) sequence that was later identified as the human homologue of rat sucrose non-fermenting protein (SNF-1) related kinase (rSNRK), a novel SNF-1-related kinase previously characterized in the rat . In the present study we report the cloning of the complete human SNF-1 related kinase (hSNRK) cDNA and show that the gene spans 39.8 kb at region 3p21 and contains six exons . Recombinant expression of the hSNRK coding sequence in Escherichia coli led to the production of a functional protein kinase of 85 kDa . Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of hSNRK expression in fetal CD34+ hematopoietic progenitors revealed its continuous expression throughout human development with higher levels in highly dividing CD34+ CD38+ cells compared to quiescent CD34+ CD38- cells . This observation, together with the expression of hSNRK in numerous human leukemic cell lines, may reflect an implication of hSNRK protein in hematopoietic cell proliferation or differentiation.In the mouse, the SNRK cDNA is 4.6-kb-long and encodes a protein of 748 amino acids with a predicted molecular mass of 81,930 Da . The proteins from human, rat and mouse are strongly conserved and are characterized by the presence of a serine/threonine kinase catalytic domain, a bipartite nuclear targeting signal and an ubiquitin-associated domain . In situ hybridization and RT-PCR analysis of the pattern of mSNRK expression in the mouse reveals that it is temporally and spatially regulated during embryogenesis, and widespread expressed in adult tissues.

J Appl Microbiol, 2002, 93(4), 668 - 74
Culture conditions influencing phytase production of Mitsuokella jalaludinii, a new bacterial species from the rumen of cattle; Lan GQ et al.; AIMS: The effects of pH, temperature, phytate, glucose, phosphate and surfactants on the phytase production of Mitsuokella jalaludinii, a new bacterial species from the rumen of cattle, were evaluated . METHOD AND RESULTS: By using batch fermentation, it was found that phytase production of M . jalaludinii was induced by sodium phytate . Inclusion of glucose into rice bran-soybean milk (RB-SM) medium inhibited the phytase production . Supplementation of 0.05-0.5% of phosphate to RB-SM medium neither inhibited nor stimulated the phytase production . The optimum pH and temperature for the phytase production of M . jalaludinii were 7.02 and 39 degrees C, respectively . Surfactants such as Tween-80 (0.5% and 1.0%) and Triton X-100 (1.0%) did not affect the phytase production . CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The study has provided important data on the culture conditions affecting phytase production of M . jalaludinii, which will facilitate future research on the optimization of fermentation processes for production of high phytase activity by M . jalaludinii.

J Appl Microbiol, 2002, 93(4), 608 - 15
Saccharomyces cerevisiae wine yeast populations in a cold region in Argentinean Patagonia . A study at different fermentation scales; Lopes CA et al.; AIMS: To study the diversity and dynamics of indigenous Saccharomyces wine populations during Malbec spontaneous fermentation, a representative Patagonian red wine, at both industrial and laboratory scale . METHODS AND RESULTS: Two molecular techniques, including restriction fragment length polymorphism of mitochondrial (mt) DNA and polymorphism of amplified delta interspersed element sequences, were used for characterization of indigenous yeasts at strain level . The mtDNA restriction patterns showed the major discriminative power; however, by combining the two molecular approaches it was possible to distinguish a larger number of strains and, therefore, draw more representative conclusions about yeast diversity . Although a great diversity of wild Saccharomyces cerevisiae strains was observed, only nine represented more than half of the total Saccharomyces yeast biota analysed; five of these were common and took over the Malbec must fermentation in both vinifications . CONCLUSIONS: Many different indigenous S . cerevisiae strains were identified; nevertheless, the dominant strains in both industrial and laboratory vinification processes were just a few and the same . SIGNIFICANCE AND IMPACT OF THE STUDY: Small-scale fermentation appears to be a valuable tool in winemaking, one especially helpful in evaluating microbiological aspects of as well as possible interactions between inoculated selected strains and native strains.

J Food Prot, 2002 Sep, 65(9), 1498 - 501
Formation of biogenic amines in a typical semihard Italian cheese; Innocente N et al.; Given that the concentration of biogenic amines in cheeses depends on variety, age, and type of microflora, a study was undertaken to investigate the formation of these compounds during the ripening of a typical semihard Italian cheese . Tryptamine, phenylethylamine, putrescine, cadaverine, histamine, and tyramine contents were calculated in 30 samples of Montasio cheese characterized by different levels of proteolysis . Histamine and tyramine were the major amines . Tryptamine and phenylethylamine concentrations were very low at all ripening periods . Putrescine and cadaverine were present only in samples with anomalous fermentation processes . The relationship between the total amine content and the proteolytic maturation coefficient was calculated; however, even for higher levels of proteolysis, the biogenic amine content in Montasio cheese was below the level considered potentially toxic.

Bioseparation, 2001, 10(4-5), 221 - 7
Kinetic studies of clavulanic acid recovery by ion exchange chromatography; Barboza M et al.; Clavulanic acid (CA) is a beta-lactamase inhibitor produced by strains of Streptomyces clavuligerus . Nowadays, the combination of CA with amoxycillin is the most successful example of the use of a beta-lactam antibiotic sensitive to beta-lactamases together with an inhibitor of these enzymes . Clavulanic acid is purified from fermentation broth by a series of steps consisting mainly of two-phase separation processes such as liquid-liquid extraction, adsorption or ion-exchange chromatography, among others . Amberlite IRA 400, a strong anion-exchange resin, has a very high adsorption capacity for CA (Mayer et al . 1997) . This resin can be pre-treated with NaCl (chloride cycle), to remove selectively only those anions, which are able to displace chloride from the resin or with NaOH (hydroxyl cycle), to remove all species of anions . In order to decide the best operating conditions for CA recovery by ion-exchange resins and then to construct a model of this separation process, batch experiments were conducted using Amberlite IRA 400 in the chloride cycle . These runs were carried out in a 200 ml stirred tank, at two different initial solution pH, 6.2 and 4.0; the temperature was maintained at 10 degrees C and 20 degrees C during adsorption and 30 degrees C during the desorption step . It was possible, on the basis of these batch results, to model the separation process, including the adsorption kinetics, equilibrium data and mass transfer limitations.

Curr Microbiol, 2002 Nov, 45(5), 340 - 5
Amino acid deamination by ruminal Megasphaera elsdenii strains; Rychlik JL et al.; When ruminal fluid from a cow fed timothy hay was serially diluted (10-fold increments into anaerobic broth containing 15 mg ml(-1) Trypticase), the low dilutions (< or =10(-6)) had optical densities greater than 2.0 and ammonia concentrations greater than 100 m M . The optical densities and ammonia concentrations of the 10(-8) and 10(-9) dilutions were very low, but large cocci were observed in the 10(-8) dilution . The large cocci were isolated and identified by 16S rDNA sequencing as Megasphaera elsdenii . The freshly isolated strain (JL1) grew well on Trypticase, but less than 4% of the amino acid nitrogen in Trypticase was converted to ammonia . Optical density and ammonia production were twice as great if Casamino acids were provided, and similar results were obtained with seven other strains (B159, AW106, YT91, LC1, T81, J1, and YZ70) . Specific activities of deamination (based on Casamino acids) of the eight strains ranged from 100 (strain JL1) to 325 (strain B159) nmol mg protein(-1) min(-1) . None of the strains could utilize branched-chain amino acids as an energy source for growth, but specific activities of branched-chain amino acid deamination ranged from 15 to 65 nmol mg protein(-1) min(-1) . All eight of the M . elsdenii strains grew well in the presence of 5 micro M monensin, and only two of the strains were strongly inhibited by 20 micro M monensin . On the basis of these results, it appears that M . elsdenii is deficient in peptidase activity and can utilize only a few amino acids . Some M . elsdenii strains produced ammonia and branched-chain volatile fatty acids nearly as fast as obligate amino acid-fermenting ruminal bacteria, but the extent of this production was at least fourfold lower . Because all of the strains could tolerate 5 micro M monensin, it is unlikely that this feed additive would significantly inhibit M . elsdenii in vivo.

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