Fermentations

J Biomol Screen, 2003 Jun, 8(3), 305 - 15
HPLC studio: a novel software utility to perform HPLC chromatogram comparison for screening purposes; Garcia JB et al.; A new tool, HPLC Studio, was developed for the comparison of high-performance liquid chromatography (HPLC) chromatograms from microbial extracts . The new utility makes it possible to create a virtual chromatogram by mixing up to 20 individual chromatograms . The virtual chromatogram is the first step in establishing a ranking of the microbial fermentation conditions based on either the area or diversity of HPLC peaks . The utility was used to maximize the diversity of secondary metabolites tested from a microorganism and therefore increase the chances of finding new lead compounds in a drug discovery program.

Nucleic Acids Res, 2003 Jul 15, 31(14), 4218 - 26
Temperature-sensitive mutation in yeast mitochondrial ribosome recycling factor (RRF); Teyssier E et al.; The yeast protein Rrf1p encoded by the FIL1 nuclear gene bears significant sequence similarity to Escherichia coli ribosome recycling factor (RRF) . Here, we call FIL1 Ribosome Recycling Factor of yeast, RRF1 . Its gene product, Rrf1p, was localized in mitochondria . Deletion of RRF1 leads to a respiratory incompetent phenotype and to instability of the mitochondrial genome (conversion to rho(-)/rho(0) cytoplasmic petites) . Yeast with intact mitochondria and with deleted genomic RRF1 that harbors a plasmid carrying RRF1 was prepared from spores of heterozygous diploid yeast . Such yeast with a mutated allele of RRF1, rrf1-L209P, grew on a non-fermentable carbon source at 30 but not at 36 degrees C, where mitochondrial but not total protein synthesis was 90% inhibited . We propose that Rrf1p is essential for mitochondrial protein synthesis and acts as a RRF in mitochondria.

EMBO J, 2003 Jul 15, 22(14), 3493 - 502
Crystal structure of the carboxyltransferase subunit of the bacterial sodium ion pump glutaconyl-coenzyme A decarboxylase; Wendt KS et al.; Glutaconyl-CoA decarboxylase is a biotin-dependent ion pump whereby the free energy of the glutaconyl-CoA decarboxylation to crotonyl-CoA drives the electrogenic transport of sodium ions from the cytoplasm into the periplasm . Here we present the crystal structure of the decarboxylase subunit (Gcdalpha) from Acidaminococcus fermentans and its complex with glutaconyl-CoA . The active sites of the dimeric Gcdalpha lie at the two interfaces between the mono mers, whereas the N-terminal domain provides the glutaconyl-CoA-binding site and the C-terminal domain binds the biotinyllysine moiety . The Gcdalpha catalyses the transfer of carbon dioxide from glutaconyl-CoA to a biotin carrier (Gcdgamma) that subsequently is decarboxylated by the carboxybiotin decarboxylation site within the actual Na(+) pump (Gcdbeta) . The analysis of the active site lead to a novel mechanism for the biotin-dependent carboxy transfer whereby biotin acts as general acid . Furthermore, we propose a holoenzyme assembly in which the water-filled central channel of the Gcdalpha dimer lies co-axial with the ion channel (Gcdbeta) . The central channel is blocked by arginines against passage of sodium ions which might enter the central channel through two side channels.

J Biol Chem, 2003 Sep 19, 278(38), 36169 - 75 Epub 2003 Jul 09.
The multifunctional isopropyl alcohol dehydrogenase of Phytomonas sp . could be the result of a horizontal gene transfer from a bacterium to the trypanosomatid lineage; Molinas SM et al.; Isopropyl alcohol dehydrogenase (iPDH) is a dimeric mitochondrial alcohol dehydrogenase (ADH), so far detected within the Trypanosomatidae only in the genus Phytomonas . The cloning, sequencing, and heterologous expression of the two gene alleles of the enzyme revealed that it is a zinc-dependent medium-chain ADH . Both polypeptides have 361 amino acids . A mitochondrial targeting sequence was identified . The mature proteins each have 348 amino acids and a calculated molecular mass of 37 kDa . They differ only in one amino acid, which can explain the three isoenzymes and their respective isoelectric points previously found . A phylogenetic analysis locates iPDH within a cluster with fermentative ADHs from bacteria, sharing 74% similarity and 60% identity with Ralstonia eutropha ADH . The characterization of the two bacterially expressed Phytomonas enzymes and the comparison of their kinetic properties with those of the wild-type iPDH and of the R . eutropha ADH strongly support the idea of a horizontal gene transfer event from a bacterium to a trypanosomatid to explain the origin of the iPDH in Phytomonas . Phytomonas iPDH and R . eutropha ADH are able to use a wide range of substrates with similar Km values such as primary and secondary alcohols, diols, and aldehydes, as well as ketones such as acetone, diacetyl, and acetoin . We speculate that, as for R . eutropha ADH, Phytomonas iPDH acts as a safety valve for the release of excess reducing power.

Rev Esp Salud Publica, 2003 May-Jun, 77(3), 317 - 31
{Functional nutrition and optimal nutrition . Near or far?}; Silveira Rodriguez MB et al.; The concept of functional food, about which scientific agreement is still lacking, springs from the field of Optimum Nutrition, aimed at modifying genetic and physiological aspects of human life and at the prevention and treatment of a growing number of diseases, far beyond merely covering nutritional requirements . From the European Union perspective, functional foods can be natural as well as industrially processed foods . The leading functional foods regarding which the soundest scientific evidence exists are probiotics, live microbial food ingredients represented mainly by fermented dairy products . Prebiotics, such as inulin-type fructans, are the trophic substrate of probiotics and potential intestinal microflora selectors . The combination of prebiotics and probiotics is termed synbiotic . Innumerable substances are known to have functional effects: soluble and insoluble fiber, phytosterols, phytoestrogens, monounsaturated and polyunsaturated fatty acids, phenol derivatives, vitamins and other phytochemicals . Functional foods exert their actions on different systems, especially the gastrointestinal, cardiovascular and immunological ones, acting too as enhancers of development and differentiation and positively modulating nutrient metabolism, gene expression, oxidative stress and the psychic sphere . The establishment of Health Claims must be firmly based upon scientific knowledge and legal regulation . Efficient biomarkers related to biological response must be found . Furthermore, it is essential to analyze possible diet or drug interactions as well as it is indispensable to conduct valid studies on humans . The prime objective must be the diet as a whole . Thus, the future challenge of a functional diet emerges.

J Biol Chem, 2003 Sep 19, 278(38), 36924 - 33 Epub 2003 Jul 07.
Tor1/2 regulation of retrograde gene expression in Saccharomyces cerevisiae derives indirectly as a consequence of alterations in ammonia metabolism; Tate JJ et al.; Retrograde genes of Saccharomyces cerevisiae encode the enzymes needed to synthesize alpha-ketoglutarate, required for ammonia assimilation, when mitochondria are damaged or non-functional because of glucose fermentation . Therefore, it is not surprising that a close association exists between control of the retrograde regulon and expression of nitrogen catabolic genes . Expression of these latter genes is nitrogen catabolite repression (NCR)-sensitive, i.e . expression is low with good nitrogen sources (e.g . glutamine) and high when only poor (e.g . proline) or limiting nitrogen sources are available . It has been reported recently that both NCR-sensitive and retrograde gene expression is negatively regulated by glutamine and induced by treating cells with the Tor1/2 inhibitor, rapamycin . These conclusions predict that NCR-sensitive and retrograde gene expression should respond in parallel to nitrogen sources, ranging from those that highly repress NCR-sensitive transcription to those that elicit minimal NCR . Because this prediction did not accommodate earlier observations that CIT2 (a retrograde gene) expression is higher in glutamine than proline containing medium, we investigated retrograde regulation further . We show that (i) retrograde gene expression correlates with intracellular ammonia and alpha-ketoglutarate generated by a nitrogen source rather than the severity of NCR it elicits, and (ii) in addition to its known regulation by NCR, NAD-glutamate dehydrogenase (GDH2) gene expression is down-regulated by ammonia under conditions where NCR is minimal . Therefore, intracellular ammonia plays a pivotal dual role, regulating the interface of nitrogen and carbon metabolism at the level of ammonia assimilation and production . Our results also indicate the effects of rapamycin treatment on CIT2 transcription, and hence Tor1/2 regulation of retrograde gene expression occur indirectly as a consequence of alterations in ammonia and glutamate metabolism.

Life Sci, 2003 Jul 25, 73(10), 1289 - 98
Dietary supplementation of fermented soybean, natto, suppresses intimal thickening and modulates the lysis of mural thrombi after endothelial injury in rat femoral artery; Suzuki Y et al.; We have previously demonstrated that natto-extracts containing nattokinase (NK) inactivates plasminogen activator inhibitor type 1 and then potentiates fibrinolytic activity . In the present study, we investigated the effects of dietary supplementation with natto-extracts on neointima formation and on thrombolysis at the site of endothelial injury . Endothelial damage in the rat femoral artery was induced by intravenous injection of rose bengal followed by focal irradiation by transluminal green light . Dietary natto-extracts supplementation containing NK of 50 or 100 CU/body was started 3 weeks before endothelial injury and then continued for another 3 weeks . Intimal thickening in animals given supplementation was significantly (P<0.01) suppressed compared with controls and the intima/media ratio in animals with 50 and 100 CU/body NK and control group was 0.09 +/- 0.03, 0.09 +/- 0.06 and 0.16 +/- 0.12, respectively . Although femoral arteries were reopened both in control animals and those treated with NK within 8 hours after endothelial injury, mural thrombi were histologically observed at the site of endothelial injury . In the control group, the center of vessel lumen was reopened and mural thrombi were attached on the surface of vessel walls . In contrast, in NK-treated groups, thrombi near the vessel wall showed lysis and most of them detached from the surface of vessel walls . In conclusion, dietary natto-extracts supplementation suppressed intimal thickening produced by endothelial injury in rat femoral artery . These effects may partially be attributable to NK, which showed enhanced thrombolysis near the vessel wall.

Metab Eng, 2003 Apr, 5(2), 86 - 95
Production process monitoring by serial mapping of microbial carbon flux distributions using a novel sensor reactor approach: I--Sensor reactor system; El Massaoudi M et al.; A novel Sensor Reactor technology is presented which permits 13C labeling experiments for metabolic flux analysis during large-scale, semi-industrial, (fed-) batch fermentation processes deriving a series of flux maps that document fermentation courses in detail . The small-scale Sensor Reactor can be inoculated within 1.50-1.20s via a special inoculation unit with an inoculation volume accuracy of 1.025+/-0.021 L . The large-scale production reactor (here: 300 L) and the Sensor Reactor were run in parallel master/slave modes to control the current pH, temperature, pressure and dissolved oxygen values as changing set points for the Sensor Reactor . Using an automated pulsing technology, glucose pulses of 5 g/L could be realized within 0.51 s . The similarity of fermentations in the Sensor Reactor with the production process was demonstrated by studying L-lysine production with C . glutamicum during multiple, 'simulated' labeling experiments each lasting 2.5h . 'Real' labeling experiments are presented in Part II.

Neural Netw, 2003 Jun-Jul, 16(5-6), 847 - 53
Electronic nose based tea quality standardization; Dutta R et al.; In this paper we have used a metal oxide sensor (MOS) based electronic nose (EN) to analyze five tea samples with different qualities, namely, drier month, drier month again over-fired, well fermented normal fired in oven, well fermented overfired in oven, and under fermented normal fired in oven . The flavour of tea is determined mainly by its taste and smell, which is generated by hundreds of Volatile Organic Compounds (VOCs) and Non-Volatile Organic Compounds present in tea . These VOCs are present in different ratios and determine the quality of the tea . For example Assamica (Sri Lanka and Assam Tea) and Assamica Sinesis (Darjeeling and Japanese Tea) are two different species of tea giving different flavour notes . Tea flavour is traditionally measured through the use of a combination of conventional analytical instrumentation and human or ganoleptic profiling panels . These methods are expensive in terms of time and labour and also inaccurate because of a lack of either sensitivity or quantitative information . In this paper an investigation has been made to determine the flavours of different tea samples using an EN and to explore the possibility of replacing existing analytical and profiling panel methods . The technique uses an array of 4 MOSs, each of, which has an electrical resistance that has partial sensitivity to the headspace of tea . The signals from the sensor array are then conditioned by suitable interface circuitry . The data were processed using Principal Components Analysis (PCA), Fuzzy C Means algorithm (FCM) . We also explored the use of a Self-Organizing Map (SOM) method along with a Radial Basis Function network (RBF) and a Probabilistic Neural Network classifier . Using FCM and SOM feature extraction techniques along with RBF neural network we achieved 100% correct classification for the five different tea samples with different qualities . These results prove that our EN is capable of discriminating between the flavours of teas manufactured under different processing conditions, viz . over-fermented, over-fired, under fermented, etc.

J Agric Food Chem, 2003 Jul 16, 51(15), 4427 - 35
Determination of tea components with antioxidant activity; Cabrera C et al.; Levels of essential elements with antioxidant activity, as well as catechins, gallic acid, and caffeine levels, in a total of 45 samples of different teas commercialized in Spain have been evaluated . Chromium, manganese, selenium, and zinc were determined in the samples mineralized with HNO(3) and V(2)O(5), using ETAAS as the analytical technique . The reliability of the procedure was checked by analysis of a certified reference material . Large variations in the trace element composition of teas were observed . The levels ranged from 50.6 to 371.4 ng/g for Cr, from 76.1 to 987.6 microg/g for Mn, from 48.5 to 114.6 ng/g for Se, and from 56.3 to 78.6 ng/g for Zn . The four major catechins {(-)-epigallocatechin gallate (EGCG), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), and (-)-epicatechin (EC)}, gallic acid (GA), and caffeine were simultaneously determined by a simple and fast HPLC method using a photodiode array detector . In all analyzed samples, EGCG ranged from 1.4 to 103.5 mg/g, EGC from 3.9 to 45.3 mg/g, ECG from 0.2 to 45.6 mg/g, and EC ranged from 0.6 to 21.2 mg/g . These results indicated that green tea has a higher content of catechins than both oolong and fermented teas (red and black teas); the fermentation process during tea manufacturing reduces the levels of catechins significantly . Gallic acid content ranged from 0.039 to 6.7 mg/g; the fermentation process also elevated remarkably gallic acid levels in black teas (mean level of 3.9 +/- 1.5 mg/g) . The amount of caffeine in the analyzed samples ranged from 7.5 to 86.6 mg/g, and the lower values were detected in green and oolong teas . This study will be useful for the appraisal of trace elements and antioxidant components in various teas, and it will also be of interest for people who like drinking this beverage.

Lipids, 2003 Apr, 38(4), 353 - 8
n-3 long-chain FA decrease serum levels of TG and remnant-like particle-cholesterol in humans; Hamazaki K et al.; A large number of papers have reported that administration of n-3 FA reduced serum TG concentrations in hypertriglyceridemic patients . However, few studies have examined the effect of n-3 FA on serum concentrations of remnant-like particle (RLP) cholesterol . Volunteers (n = 41) whose serum TG concentrations were 100-300 mg/dL were recruited and randomly assigned to either an n-3 FA group or a control group with stratification by sex, age, and serum TG level in a double-blind manner . The subjects in the n-3 FA group were administered 125 mL of fermented soybean milk with fish oil containing 600 mg of EPA and 260 mg of DHA/d for 12 wk . The controls consumed control soybean milk with olive oil . Fasting blood samples were obtained before the start of administration and at 4, 8, and 12 wk . EPA concentrations in red blood cells increased significantly in all but one subject in the n-3 FA group, with no significant changes in the control group . TG levels decreased more in the n-3 FA group than in the control group at weeks 4 (P < 0.05), 8 (P < 0.01), and 12 (P < 0.05) with their baseline as covariate . RLP cholesterol levels decreased more in the n-3 FA group than in the control at weeks 8 (P < 0.01) and 12 (P < 0.05) with their baseline as covariate . The groups did not differ in the other lipid levels . It is likely that n-3 long-chain FA may exert anti-atherosclerotic effects by lowering serum TG and RLP-cholesterol levels even at the dose of 860 mg/d.

Neurogastroenterol Motil, 2003 Aug, 15(4), 427 - 33
Abdominal distension after colonic lactulose fermentation recorded by a new extensometer; Basilisco G et al.; Colonic lactulose fermentation induces bloating, but whether it also causes abdominal distension is not known . The aim of this study was to assess the effect of colonic lactulose fermentation on abdominal girth using a new extensometer . We recorded abdominal girth in 24 healthy subjects by means of an extensometer that measures the phase shift of an ultrasound wave propagating in a tube encircling the abdomen . The recordings were continuously made for 3 h after the ingestion of 100 mL of tap water with (16 subjects) or without (eight subjects) 10 g of lactulose . Every 10 min, H2 in the breath was analysed and the intensity of bloating was recorded . Bloating was never reported after water ingestion, whereas it was reported by 10 subjects after lactulose ingestion (P = 0.002) . The mean +/- SD changes in abdominal girth in comparison with resting conditions were statistically significant after lactulose ingestion (3 +/- 2.9 mm; P = 0.002) but not after water ingestion (-0.2 +/- 2.7 mm; P = 0.82) . The area under the curve of the changes in abdominal girth after lactulose were significantly greater than after water ingestion (P = 0.03) . In conclusion, colonic lactulose fermentation induces bloating and abdominal distension . The new extensometer is useful for continuously recording changes in abdominal girth.

Diabetes Nutr Metab, 2003 Apr, 16(2), 94 - 101
Oligofructose does not affect the development of type 1 diabetes mellitus induced by dietary proteins in the diabetes-prone BB rat model; Perrin IV et al.; BACKGROUND: Prevention of Type 1 diabetes mellitus (T1DM), a major childhood chronic disease with rapidly increasing incidence, is an urgent topic of research . We investigated whether 5% oligofructose (OF) as compared to 5% cellulose had a protective effect against diet-induced T1DM in the diabetes-prone BioBreeding (BB) rat model . METHODS: Groups of BB rats were fed the experimental diets from weaning . The diets were a cereal-based rodent diet (diabetogenic, positive control) and semi-synthetic rodent diets containing hydrolysed casein (non-diabetogenic, negative control), soy or whey as the sole protein source and 5% cellulose as fibre source . In additional groups fed soy and whey protein, the fibre source was 5% OF . T1DM incidence up to the age of 160 days was recorded applying biochemical and morphological criteria . Physiological effects of fibre were assessed through the analysis of biochemical parameters in plasma and of the protein/DNA ratio in intestinal mucosa . RESULTS: T1DM incidence was diet-dependent . Cereal-, soy- and whey-based diets were significantly more diabetogenic than the hydrolysed casein-based diet . Five per cent OF did not affect the incidence of T1DM induced by either soy or whey proteins as compared to cellulose, nor induce any of the biological effects attributed to a fermentable fibre . CONCLUSIONS: In the BB rat model, 5% OF in the diet did not have any protective effects against diet-induced T1DM . The present data do not suggest dietary OF as a promising approach for the dietary prevention of T1DM.

Appl Microbiol Biotechnol, 2003 Oct, 62(5-6), 474 - 7 Epub 2003 Jul 04.
Biorefinery; Ohara H; The biorefinery produces fuels, solvents, plastics and food for human beings . In some countries, these biorefinery products are made from waste biomass . The main processes in the biorefinery involve ethanol fermentation and lactic acid fermentation . For the biorefinery, many hybrid technologies were developed from different fields, such as bioengineering, polymer chemistry, food science and agriculture.

Curr Genet, 2003 Oct, 44(1), 49 - 57 Epub 2003 Jul 05.
Loss of function of the Fusarium oxysporum SNF1 gene reduces virulence on cabbage and Arabidopsis; Ospina-Giraldo MD et al.; Fusarium oxysporum pathogenicity is believed to require the activity of cell wall-degrading enzymes . Production of these enzymes in fungi is subject to carbon catabolite repression, a process that in yeast is mostly controlled by the SNF1 (sucrose non-fermenting 1) gene . To elucidate the role of cell wall-degrading enzymes in F . oxysporum pathogenicity, we cloned and disrupted its SNF1 homologue ( FoSNF1) . The fosnf1 mutants had a reduced expression of several genes encoding cell wall-degrading enzymes and grew poorly on certain carbon sources . Infection assays on Arabidopsis thaliana and Brassica oleracea revealed that progression of wilt symptoms in plants infected by fosnf1 mutants was considerably delayed, in comparison with those infected by a wild-type strain . In conclusion, mutations in FoSNF1 prevent F . oxysporum from properly derepressing the production of cell wall-degrading enzymes, compromise the utilization of certain carbon sources, and reduce its virulence on A . thaliana and B . oleracea.

Br J Nutr, 2003 Jul, 90(1), 75 - 85
Restoration of the integrity of rat caeco-colonic mucosa by resistant starch, but not by fructo-oligosaccharides, in dextran sulfate sodium-induced experimental colitis; Moreau NM et al.; Butyrate is recognised as efficient in healing colonic inflammation, but cannot be used as a long-term treatment . Dietary fibre that produces a high-butyrate level when fermented represents a promising alternative . We hypothesised that different types of dietary fibre do not have the same efficiency of healing and that this could be correlated to their fermentation characteristics . We compared short-chain fructo-oligosaccharides (FOS) and type 3 resistant starch (RS) in a previously described dextran sulfate sodium (DSS)-induced colitis model . Seventy-two Sprague-Dawley rats received water (control rats) or DSS (50 g DSS/l for 7 d then 30 g DSS/l for 7 (day 7) or 14 (day 14) d) . The rats were fed a basal diet (BD), or a FOS or RS diet creating six groups: BD-control, BD-DSS, FOS-control, FOS-DSS, RS-control and RS-DSS . Caeco-colonic inflammatory injuries were assessed macroscopically and histologically . Short-chain fatty acids (SCFA) were quantified in caeco-colon, portal vein and abdominal aorta . At days 7 and 14, caecal and distal macroscopic and histological observations were improved in RS-DSS compared with BD-DSS and also with FOS-DSS rats . Caeco-colonic SCFA were reduced in FOS-DSS and RS-DSS groups compared with healthy controls . The amount of butyrate was higher in the caecum of the RS-DSS rats than in the BD-DSS and FOS-DSS rats, whereas distal butyrate was higher in FOS-DSS rats . Partially explained by higher luminal levels of SCFA, especially butyrate, the healing effect of RS confirms the involvement of some types of dietary fibre in inflammatory bowel disease . Moreover, the ineffectiveness of FOS underlines the importance of the type of dietary substrate.

J Ind Microbiol Biotechnol, 2003 Jul, 30(7), 440 - 4 Epub 2003 Jul 03.
Role of the pectinolytic enzyme in the lactic acid fermentation of potato pulp by Rhizopus oryzae; Saito K et al.; Rhizopus oryzae strain NBRC 4707 produced lactic acid and ethanol more efficiently than strain NRRL 395 in potato pulp, an agricultural by-product of the starch industry . The two strains developed comparable activities of xylanase, cellulase, alpha-amylase, and glucoamylase, while the polygalacturonase activity of strain NBRC 4707 was double that of strain NRRL 395 . The addition of commercial pectinase enhanced the formation of metabolites, suggesting that the degradation of pectic substances determines the fermentation of potato pulp by R . oryzae . Orange and apple peel were more effective in the induction of polygalacturonase activity than potato pulp, sugarbeet pulp, or wheat bran when used as a principal carbon source for fungal growth in a solid-state culture . The fungal cells in both types of fruit peel stimulated the fermentation of potato pulp and increased the quantity of lactic acid and ethanol to higher levels than those in other agricultural by-products.

Biosci Biotechnol Biochem, 2003 Jun, 67(6), 1278 - 83
Angiotensin I-converting enzyme inhibitory peptides isolated from tofuyo fermented soybean food; Kuba M et al.; Angiotensin I-converting enzyme (ACE) inhibitory activity was observed in a tofuyo (fermented soybean food) extract with an IC(50) value of 1.77 mg/ml . Two ACE inhibitors were isolated to homogeneity from the extract by adsorption and gel filtration column chromatography, and by reverse-phase high-performance liquid chromatography (HPLC) . The purified substances reacted with 2,4,6-trinitrobenzensulfonic acid sodium salt . The amino acid sequences of these inhibitors determined by Edman degradation were Ile-Phe-Leu (IC(50), 44.8 microM) and Trp-Leu (IC(50), 29.9 microM) . The Ile-Phe-Leu sequence is found in the alpha- and beta-subunits of beta-conglycinin, while the Trp-Leu sequence is in the B-, B1A- and BX-subunits of glycinin from soybean . Both of the peptides are non-competitive inhibitors . The inhibitory activity of Trp-Leu was completely preserved after a treatment with pepsin, chymotrypsin or trypsin . Even after successive digestion by these gastrointestinal proteases, the activity remained at 29% of the original value.

Appl Microbiol Biotechnol, 2003 Jul, 62(1), 92 - 8 Epub 2003 Feb 26.
Glucose oxidation by Gluconobacter oxydans: characterization in shaking-flasks, scale-up and optimization of the pH profile; Silberbach M et al.; In this study, the advantage of a novel measuring device for the online determination of oxygen and carbon dioxide transfer rates in shaking-flasks is reported for glucose oxidation by Gluconobacter oxydans . In this fermentation process, this device was used for the characterization of the oxidation pattern of different strains . G . oxydans NCIMB 8084 forms 2,5-diketogluconate from d-glucose in a multi-stage process via three different membrane-bound dehydrogenases . This strain was chosen for a scale-up of the process from shaking-flasks to a 2-l stirred vessel . An enhancement of 2,5-diketogluconate production was realized by controlling the pH at different levels during the fermentation.

Appl Microbiol Biotechnol, 2003 Jul, 62(1), 76 - 82 Epub 2003 Feb 26.
The effect of acetate pathway mutations on the production of pyruvate in Escherichia coli; Tomar A et al.; We compared pyruvate accumulation in six strains of Escherichia coli and their corresponding ppc mutants . Each strain contained a mutation of a gene involved in the pathway to acetate synthesis . Strains with mutations in genes encoding the pyruvate dehydrogenase complex generally exhibited the greatest pyruvate accumulation of which CGSC6162 (an aceF mutant) and CGSC6162 Delta ppc were studied in greater detail in controlled fermenters . Both CGSC6162 and CGSC6162 Delta ppc accumulated greater than 35 g/l pyruvate in a medium supplemented with acetate . We observed pyruvate mass yields from glucose of 0.72 in CGSC6162, with volumetric productivities above 1.5 g l(-1) h(-1) . For CGSC6162 Delta ppc, we observed pyruvate yields of 0.78 and volumetric productivities above 1.2 g l(-1) h(-1) . CGSC6162 consumed all initially supplied acetate, while CGSC6162 Delta ppc first consumed and then generated acetate during the course of a 36 h fermentation . Acetate generation and pyruvate oxidase activity was pH- and temperature-dependent, with a pH of 7.0 and the lowest temperature studied (32 degrees C) favoring the greatest pyruvate generation . Lactate was an unexpected by-product even though measured lactate dehydrogenase (LDH) activity was very low.

Appl Microbiol Biotechnol, 2003 Jul, 62(1), 35 - 40 Epub 2003 Mar 13.
Kinetics of L-lysine fermentation: a continuous culture model incorporating oxygen uptake rate; Ensari S et al.; For process design and optimization, it is essential to have a mathematical model that represents the system well . Many past studies do not go beyond empirically fitting experimental data . In the present study, an unstructured model incorporating oxygen uptake and dissolved oxygen concentration was developed for a continuous culture of L-lysine . Specific rate expressions of cell growth, substrate consumption, product formation, and oxygen uptake were developed and incorporated in the model . The model predicts very well the effects of operational parameters, such as the dilution rate and the feed substrate concentration . It is also able to predict the unsteady-state dynamics of continuous L-lysine fermentation.

Proc Natl Acad Sci U S A, 2003 Jul 8, 100(14), 8298 - 303 Epub 2003 Jun 30.
Complete genome sequence of the marine planctomycete Pirellula sp . strain 1; Glockner FO et al.; Pirellula sp . strain 1 ("Rhodopirellula baltica") is a marine representative of the globally distributed and environmentally important bacterial order Planctomycetales . Here we report the complete genome sequence of a member of this independent phylum . With 7.145 megabases, Pirellula sp . strain 1 has the largest circular bacterial genome sequenced so far . The presence of all genes required for heterolactic acid fermentation, key genes for the interconversion of C1 compounds, and 110 sulfatases were unexpected for this aerobic heterotrophic isolate . Although Pirellula sp . strain 1 has a proteinaceous cell wall, remnants of genes for peptidoglycan synthesis were found . Genes for lipid A biosynthesis and homologues to the flagellar L- and P-ring protein indicate a former Gram-negative type of cell wall . Phylogenetic analysis of all relevant markers clearly affiliates the Planctomycetales to the domain Bacteria as a distinct phylum, but a deepest branching is not supported by our analyses.

Bioresour Technol, 2003 Oct, 90(1), 95 - 100
Ethanol fermentation of acid-hydrolyzed cellulosic pyrolysate with Saccharomyces cerevisiae; Yu Z et al.; The acid hydrolysis of cellulosic pyrolysate to glucose and its fermentation to ethanol were investigated . The maximum glucose yield (17.4%) was obtained by the hydrolysis with 0.2 mol sulfuric acid per liter pyrolysate using autoclaving at 121 degrees C for 20 min . The fermentation by Saccharomyces cerevisiae of a hydrolysate medium containing 31.6 g/l glucose gave 14.2 g/l ethanol in 24 h, whereas the fermentation of the medium containing 31.6 g/l pure glucose gave 13.7 g/l ethanol in 18 h . The results showed that the acid-hydrolyzed pyrolysate could be used for ethanol production . Different nitrogen sources were evaluated and the best ethanol concentration (15.1 g/l) was achieved by single urea . S . cerevisiae (R) was obtained by adaptation of S . cerevisiae to the hydrolysate medium for 12 times, and 40.2 g/l ethanol was produced by S . cerevisiae (R) in the fermentation with the hydrolysate medium containing 95.8 g/l glucose, which was about 47% increase in ethanol production compared to its parent strain.

Bioresour Technol, 2003 Oct, 90(1), 89 - 94
Effect of saponification on the anaerobic digestion of solid fatty residues; Mouneimne AH et al.; In France, fatty residues considered as "non-ultimate" waste will not be accepted in landfill from 2002, in accordance with French legislation . Anaerobic digestion appears as an alternative process to mobilize and profitably use such fermentable waste . In this work, the effect of an alkaline pretreatment on the degradation of hexane extractible matter (HEM) and the production of volatile fatty acids (VFAs) was compared in reactors working at pH 6.5 and 8.5 . The results obtained showed that 40% (+/- 0.1) of HEM were degraded at pH 8.5 versus 10% (+/- 0.3) at pH 6.5, regardless of the alkali agent used to saponify the greasy wastes . The highest performances of VFA production (8.45% +/- 0.3) were obtained at pH 8.5 with greases saponified by potassium hydroxide, compared to results (4.25% +/- 0.1) with greasy waste saponified by sodium hydroxide . This difference in VFA production might be attributable to biotoxic molecules generated during the saponification of greases by soda, limiting consequently the VFA production process.

Bioresour Technol, 2003 Oct, 90(1), 59 - 63
Effect of select nitrocompounds on ruminal fermentation; an initial look at their potential to reduce economic and environmental costs associated with ruminal methanogenesis; Anderson RC et al.; Methane production by ruminal microbes during the digestion of feedstuffs is an inefficient process resulting in losses of 2-12% of the gross energy consumed by ruminants . Presently, we report the effect of three inhibitors on ruminal methane production in vitro . Mixed populations of ruminal microbes collected from cannulated cows maintained on an alfalfa hay:corn diet (50:50) were incubated at 39 degrees C for 24 h under a 100% carbon dioxide gas phase in closed tubes with 72 mM added sodium formate . Cultures were supplemented with 12 mM 2-nitropropanol, nitroethane or nitroethanol (experiment 1) or with 2, 12 or 24 mM nitroethane or a combination of 12 mM nitroethane and 4 mM nitroethanol (experiment 2) . Control cultures containing no added nitrocompound were incubated simultaneously with treated incubations . Methane concentrations were reduced (P<0.05) from those measured in control incubations (27.6 +/- 2.1 and 17.7 +/- 0.8 micromol/ml; mean +/- SD for experiments 1 and 2, respectively) by at least 57% and as much as 94% in the nitrocompound supplemented incubations . By comparison, the widely fed methane inhibitor, monensin, typically reduces ruminal methane production by about 33% . Concentrations of volatile fatty acids and ammonia that accumulated in the nitrocompound supplemented incubations were not markedly affected compared to those produced by control cultures despite the reductions in methane produced . Hydrogen accumulated only slightly in cultures supplemented with the nitrocompounds . These results demonstrate that 2-nitropropanol, nitroethane and nitroethanol inhibit ruminal methane production . Further research is warranted to determine the mechanisms responsible for this inhibition and to see if these inhibitors can be used in practical application to reduce economic and environmental costs associated with ruminal methanogenesis.

Biosci Biotechnol Biochem, 2003 May, 67(5), 1018 - 23
The production of a new tempeh-like fermented soybean containing a high level of gamma-aminobutyric acid by anaerobic incubation with Rhizopus; Aoki H et al.; A cultivation procedure for the preparation of a new tempeh-like fermented soybean containing a high level of gamma-aminobutyric acid was developed . Steamed soybeans were incubated aerobically with Rhizopus microsporus var . oligosporus IFO 8631 for 20 h, and then anaerobically incubated for 5 h by replacement of the atmosphere with nitrogen . The GABA content in the aerobically fermented soybeans was about 30 mg per 100 g dry fermented soybeans, while the anaerobically cultivation was about 370 mg/100 g dry fermented soybeans . The incubation with several strains of Rhizopus species showed that all of R . microsporus var . oligosporus and R . oryzae examined accumulated GABA in the anaerobically fermented soybeans . In particular, R . microsporus var . oligosporus IFO 32002 and IFO 32003 showed the highest content of GABA (1,740 mg/100 g dry fermented soybeans and 1,500 mg/100 g dry fermented soybeans, respectively) . Moreover, the free protein amino acids increased greatly in the fermented soybeans during the anaerobic cultivation.

Vet Rec, 2003 Jun 14, 152(24), 735 - 9
Effect of concentrate composition and cubicle bedding on the development of hoof haemorrhages in Holstein heifers after calving; Livesey CT et al.; Diseases of the hoof horn are the most common cause of lameness in the UK dairy herd, and diet, particularly starch intake, is commonly implicated in their development . This study assessed whether replacing starch carbohydrate with sugar beet pulp, in a diet containing 50 per cent of the dry matter as forage, would significantly reduce the development of hoof horn haemorrhages in dairy heifers after calving . It also investigated the effect of replacing wood shavings as cubicle bedding with chopped straw . The heifers fed the high-fibre diet had a significantly lower yield of milk protein and a significantly higher yield of milk fat than the heifers fed the high-starch diet, indicating that the change of diet had affected rumen fermentation . However, the change had no significant effect on the development of hoof horn haemorrhages . The change of bedding material also had no significant effect on the development of hoof horn haemorrhages.

Biochim Biophys Acta, 2003 Jun 27, 1613(1-2), 57 - 71
Fps1p channel is the mediator of the major part of glycerol passive diffusion in Saccharomyces cerevisiae: artefacts and re-definitions; Oliveira R et al.; Glycerol has been shown to cross the plasma membrane of Saccharomyces cerevisiae through (1) a H(+)/symport detected in cells grown on non-fermentable carbon sources, (2) the constitutively expressed Fps1p channel and (3) by passive diffusion . The Fps1p channel has been named a facilitator for mediating glycerol low affinity transport of the facilitated diffusion type . We present experimental evidence that this kinetic is an artefact created by glycerol kinase activity . Instead, the channel is shown to mediate the major part of glycerol's passive diffusion . This is not incompatible with Fps1p's major role in vivo, which has been previously shown to be the control of glycerol export under osmotic stress or in reaction to turgor changes . We also verified that FPS1 overexpression caused an increase in H(+)/symport V(max) . Furthermore, yfl054c and fps1 mutants were equally affected by exogenously added ethanol, being the correspondent passive diffusion stimulated . For the first time, to our knowledge, a phenotype attributed to the functioning of YFL054c gene is presented . Glycerol passive diffusion is thus apparently channel-mediated . This is discussed according to glycerol's chemical properties, which contradict the widely spread concept of glycerol's liposoluble nature . The discussion considers the multiple roles that the intracellular levels of glycerol and its pathway regulation might play as a central key to metabolism control.

J Nutr Biochem, 2003 May, 14(5), 259 - 65
Pancreatic lipase activity as influenced by unconjugated bile acids and pH, measured in vitro and in vivo; Knarreborg A et al.; The relation between pancreatic lipase activity, unconjugated bile acids and pH was studied in vitro and in vivo . Lipase activity was assayed in vitro using automatic titration, where the fatty acids liberated from the hydrolysis of glycerol tributyrate (GTB) were measured . The lipase activity was determined at different ratios of conjugated to unconjugated bile acids (100:0, 75:25, 50:50, 25:75, 0:100) in response to pH 6.6, 6.8, 7.0 and 7.5 . The in vivo study involved 96 one-day-old male broiler chickens . The chickens were assigned randomly, in pens of six animals, into two dietary treatments (8 replicate blocks), composing a non-supplemented diet (A(-)) and a diet supplemented (A(+)) with avilamycin (10 mg/kg feed) and salinomycin (40 mg/kg feed) . After 35 days, the chickens were killed and content of the proximal part of the small intestine was collected and analyzed for bacterial counts, pH, bile acid concentration, and lipase activity . Evidence for a significant pH-dependent inhibition of lipase activity by unconjugated bile acids was provided in vitro and confirmed in vivo . Due to a reduction in nutrient fermentation, the pH in the small intestine of antibiotic-fed chickens was significantly higher than in chickens fed the non-supplemented diet . The high pH in the small intestine of chickens fed the A(+)diet was accompanied by a significant increase in lipase activity, and coincided with a significantly lower concentration of unconjugated bile acids and a higher ratio of conjugated to unconjugated bile acids . This study emphasizes the important influence of unconjugated bile acids on lipase activity at physiological pH-values.

Acta Vet Scand, 2002, 43(4), 203 - 10
Persistence of Trichinella spiralis in rat carcasses experimentally mixed in different feed; Oivanen L et al.; Trichinella spiralis infected rat carcasses were incubated for 6 weeks in several animal feeds to assess how long Trichinella can present a risk for an outbreak in contaminated feeds . In groups of 6, 24 infected target rats were placed in silage, grained barley, propionic acid-preserved feed, and also into simulated pasture conditions . Test environments were sampled after one-, 2-, 4-, and 6-week-incubations . Trichinella larvae were recovered by digestion, and their infectivity was evaluated in rats . A two-week incubation reduced the number of recovered larvae, but still after 6 weeks low numbers were isolated from all feeds except from the experimental group simulating pasture conditions . After 2 weeks storage, the larvae were infective in all storage environments . However, up to 4 weeks, they survived only in the propionic acid-fermented feed and there in small numbers with reduced reproductive capability . This indicates the possibility of farm animals to get infection from rats or other infected material being hazardously mixed with hay or other feed . If silage is stored for at least one month before use, however, the risk from this forage appears to be minimized.

FEMS Microbiol Lett, 2003 Jun 27, 223(2), 227 - 30
Members of the genus Arthrobacter grow anaerobically using nitrate ammonification and fermentative processes: anaerobic adaptation of aerobic bacteria abundant in soil; Eschbach M et al.; Members of the genus Arthrobacter are usually regarded as obligate aerobic bacteria . The anaerobic growth and energy metabolism of two Arthrobacter species were investigated . Arthrobacter globiformis utilized both nitrate ammonification and lactate, acetate and ethanol producing fermentation processes for anaerobic growth . Only nitrate supported anaerobic growth of Arthrobacter nicotianae . Anaerobically induced respiratory nitrate reductase activity was detected in both strains . Neither of the tested strains used the alternative electron acceptors fumarate, dimethylsulfoxide or trimethylamine-N-oxide.

J Nat Prod, 2003 Jun, 66(6), 829 - 37
Pochonins A-F, new antiviral and antiparasitic resorcylic acid lactones from Pochonia chlamydosporia var . catenulata; Hellwig V et al.; Monorden (1) and the novel resorcylic acid lactones pochonins A (2), B (4), C (6), D (7), and E (8) as well as tetrahydromonorden (5) and pseurotin A (22) were isolated from cultures of the clavicipitaceous hyphomycete Pochonia chlamydosporia var . catenulata strain P 0297 . Fermentation of P 0297 in bromide-containing culture media led to a shift in secondary metabolite production and yielded monocillins III (3) and II (9) as major metabolites besides monorden (1) as well as the novel compounds pochonin F (10) and a monocillin II glycoside (11) as minor metabolites . Most of these compounds showed moderate activities in a cellular replication assay against Herpes Simplex Virus 1 (HSV1) and against the parasitic protozoan Eimeria tenella . In contrast to the structurally related zearalenone derivatives none of the metabolites of strain P 0297 were found to be active in a fluorescence polarization assay for determination of modulatory activities on the human estrogenic receptor ERbeta . Beta-zearalenol (17), but not zearalenone (15) and alpha-zearalenol (16), showed antiherpetic effects . We report the production, isolation, and structure elucidation of compounds 1-11 and their biological characterization.

Nutr Rev, 2003 May, 61(5 Pt 2), S40 - 8
Carbohydrate and the regulation of blood glucose and metabolism; Wolever TM; Classifying the glycemic responses of carbohydrate foods using the glycemic index (GI) requires standardized methodology for valid results . Dietary carbohydrates influence metabolism by at least four mechanisms: nature of the monosaccharides absorbed, amount of carbohydrate consumed, rate of absorption, and colonic fermentation . Reducing glycemic responses by reducing carbohydrate intake increases postprandial serum free-fatty acids (FFA) and does not improve overall glycemic control in diabetic subjects . By contrast, low-GI diets reduce serum FFA and improve glycemic control . Thus, current evidence supports FAO/WHO recommendations to maintain a high-carbohydrate diet and choose low-GI starchy foods.

Biotechnol Bioeng, 2003 Sep 5, 83(5), 503 - 13
Substrate aggregation due to aerial hyphae during discontinuously mixed solid-state fermentation with Aspergillus oryzae: experiments and modeling; Schutyser MA et al.; Solid-state fermentation (SSF) is prone to process failure due to channeling caused by evaporative cooling and the formation of an interparticle mycelium network . Mixing is needed to break the mycelium network and to avoid such failure . This study presents the first attempt to quantify and predict the effect of mycelium bonds on particle mixing and vice versa . We developed a novel experimental set-up to measure the tensile strength of hyphal bonds in SSF: Aspergillus oryzae was cultivated between two wheat-dough disks and the tensile strength of the aerial mycelium was measured with a texture analyzer . Tensile strength at different incubation times was related to oxygen consumption, to allow a translation to a rotating drum with A . oryzae cultivated on wheat grain . We performed several discontinuously mixed solid-state fermentations in the drum fermentor and measured the number and size of grain-aggregates remaining after the first mixing action . We integrated data on mycelium tensile strength into a previously developed two-dimensional discrete-particle model that calculates forces acting on individual substrate particles and the resulting radial-particle movements . The discrete-particle model predicted the quantity and size of the aggregates remaining after mixing successfully . The results show that the first mixing event in SSF with A . oryzae is needed to break mycelium to avoid aggregate formation in the grain bed, and not to distribute water added to compensate for evaporation losses, or smooth out temperature gradients .

Cesk Slov Oftalmol, 2003 Mar, 59(2), 91 - 7
{Duo-Bonharen--a new viscoelastic preparation . Experimental study}; al-Farzai A et al.; The paper discusses preclinical tests of a new viscoelastic preparation for ophthalmosurgery . Experiments revealed that preparation Duo-Bonharen which contains hyaluronic acid prepared by bacterial fermentation is suited for clinical tests . After a 24-hour interval following administration into the anterior chamber it does not raise intraocular pressure . In experiments it has no side-effects on the eye . The dispersed form in a higher concentration (5%) adheres very well to ocular tissues and protects them adequately during surgery . The cohesive form in an optimal concentration (1.5%) can be rapidly removed from inside the eye . The properties of Duo-Bonharen in experiments are comparable with similar viscomaterials which are used . The new viscoelastic material Duo-Bonharen appears suitable for clinical tests in particular for cataract surgery.

Intern Med J, 2003 Jul, 33(7), 291 - 6
Dietary fibre: a roughage guide; James SL et al.; The concept of dietary fibre is a complex one that incorporates the physical and physiological functions of fibre and its effects both systemically and local to the gastro-intestinal tract . Dietary fibre can be usefully classified according to its solubility and fermentability, which allows rational clinical application . Fibres may act in several ways including by gel-forming effects in the stomach and small intestine, by its fermentation by colonic bacteria, by a 'mop and sponge' effect, and via concomitant changes in other aspects of the diet . These actions lead to potentially beneficial effects in the gastro-intestinal tract and systemically, such as lowering serum cholesterol and improving glycaemic control . Dietary fibre has been implicated in multiple clinical situations but, although an extensive literature on putative actions and proposed physiological bases is available, high-level evidence of efficacy is limited . Nevertheless, encouraging the intake of a high-fibre diet is likely to have a range of health benefits and physicians are encouraged to follow simple practical guidelines in their everyday practice.

J Agric Food Chem, 2003 Jul 2, 51(14), 4133 - 8
The role of packaging film permselectivity in modified atmosphere packaging; Al-Ati T et al.; Modified atmosphere packaging (MAP) is commercially used to increase the shelf life of packaged produce by reducing the produce respiration rate, delaying senescence, and inhibiting the growth of many spoilage organisms, ultimately increasing product shelf life . MAP systems typically optimize O(2) levels to achieve these effects while preventing anaerobic fermentation but fail to optimize CO(2) concentrations . Altering film permselectivity (i.e., beta, which is the ratio of CO(2)/O(2) permeation coefficients) could be utilized to concurrently optimize levels of both CO(2) and O(2) in MAP systems . We investigated the effect of modifying film permselectivity on the equilibrium gas composition of a model MAP produce system packaged in containers incorporating modified poly(ethylene) ionomer films with CO(2)/O(2) permselectivites between 4-5 and 0.8-1.3 . To compare empirical to calculated data of the effect of permselectivity on the equilibrium gas composition of the MAP produce system, a mathematical model commonly used to optimize MAP of respiring produce was applied . The calculated gas composition agreed with observed values, using empirical respiration data from fresh cut apples as a test system and permeability data from tested and theoretical films . The results suggest that packaging films with CO(2)/O(2) permselectivities lower than those commercially available (<3) would further optimize O(2) and CO(2) concentration in MAP of respiring produce, particularly highly respiring and minimally processed produce.

J Agric Food Chem, 2003 Jul 2, 51(14), 4084 - 8
Adsorption of anthocyanins by yeast cell walls during the fermentation of red wines; Morata A et al.; This paper reports the anthocyanin adsorption profiles of the cell walls of different Saccharomyces strains isolated from grapes collected in the Spanish appellation controlee regions of La Rioja, Navarra, and Ribera del Duero . These strains are habitually used in red wine-making . The acyl derivatives of anthocyanins (acetyl and p-coumaryl compounds) were more strongly adsorbed than nonacyl derivatives . Peonidin-3G was also strongly adsorbed, as were its acyl derivatives . The greater presence of acetyl derivatives in the cell wall adsorbate leads to an increase in yellow color and a reduction in blue color with respect to the corresponding wine.

Nat Prod Res, 2003 Aug, 17(4), 269 - 74
Microbial transformation of the phthalideisoquinoline alkaloid, (-)-beta-hydrastine; Herath WH et al.; The phthalideisoquinoline alkaloid (-)-beta-hydrastine is one of the main active constituents of the medicinal plant, Hydrastis canadensis, which is used in many dietary supplements intended to enhance the immune system . Treatment of hydrastine with the fermentation broth of Polyporous brumalis (ATCC 34487) as a model for mammalian metabolism, gave a new alkaloid, (1S)-hydroxyhydrastine . Structure elucidation was based primarily on NMR and chiroptical studies.

Klin Lab Diagn, 2003 May, (5), 8 - 10
{Cardiac markers in the pericardial fluid in sudden coronary death}; Berestovskaia VS et al.; The pericardial fluid was examined in 26 patients without morphological signs of severe damage to cardiac histiocytes, who died unexpectedly from ischemic heart disease (IHD)--main group . The control group comprised 26 persons, who died from other (not heart diseases-asphyxia, acute blood loss, crania-cerebral trauma) . The mean age of the died was 57.4 +/- 1.5 years in the main group and 51.8 +/- 2.7 years in the control group . Cardiac markers were examined in the pericardial fluid of the died in both groups, i.e . the activity of aspartate aminotransferase (AsAT), of creatine kinase (CK), of isoenzyme KK-MB, of lactate dehydrogenase (LDG), and its isoenzyme spectrum, and, finally, the content of the cardiac troponin I (cTnI) . The statistically reliable differences were found between the two groups according to the activity of AsAT, LDG, its isoenzyme spectrum and the cTnI content . Isoenzymes LDG1 and LDG2 constituted up to 60% of the LDG activity in the pericardial fluid of those who unexpectedly died from IHD . As for the control group, the LDG activity was virtually evenly distributed between all isoenzymes . No differences were found in the activity of CK and isoenzyme KK-MB between the main and control groups . Thus, the obtained data are indicative of the "cardiac" origin of enzymes in the pericardial fluid . Finally, a number of assumptions were put forward on mechanisms of hyper-fermentation in the ischemic damage of the cardiac muscle.

Protein Expr Purif, 2003 Jul, 30(1), 62 - 8
Purification and characterization of recombinant Streptomyces griseus aminopeptidase; Ni SX et al.; Recombinant Streptomyces griseus aminopeptidase (SGAP) was produced using Cangene's expression system, CANGENUS . This heat-stable aminopeptidase with an N-terminal Ala-Pro-Asp-Ile-Pro-Leu-Ala-Asn-Val-Lys-Ala sequence was purified from 16L of Streptomyces lividans fermentation supernatant with high purity and 19.5% recovery rate . This was achieved by the combination of hydrophobic-interaction and size-exclusion chromatographic procedures . The calcium-activated zinc metalloprotein demonstrated no loss of activity at -20 degrees C for at least 8 weeks in both liquid and freeze-dried formulations . The recombinant SGAP showed an apparent molecular mass of 31 kDa by sodium dodecyl sulfate/polyacrylamide gel electrophoresis and 26.8 kDa by gel filtration . The simple, high-yield, inexpensive purification method with few intermediate steps provides a novel and practical procedure for large-scale production of active recombinant S . griseus aminopeptidase.

Chemosphere, 2003 Aug, 52(7), 1259 - 62
Chemical evidence and risks associated with soybean and rapeseed meal fermentation; Sturaro A et al.; The storage and transport of cereals and foodstuffs present recurrent problems . They may be attacked by insects or, under certain conditions, they may undergo spontaneous fermentation . Insect attack is normally avoided by fumigants, while fermentation, which depends on parameters such as temperature and humidity, is more difficult to stop and can produce chemical compounds which irremediably modify the nutritional and compositional properties of foodstuff . This paper describes the main chemical compounds produced by fermentation and self-ignition of soybean and rapeseed meal . Reported cases occurred in a storage site and during transport by ship, respectively.

Anticancer Res, 2003 Mar-Apr, 23(2B), 1605 - 12
Changes in telomerase activity, expression and splicing in response to differentiation of normal and carcinoma colon cells; Fajkus J et al.; BACKGROUND: Telomerase, a ribonucleoprotein complex catalysing synthesis of telomeric DNA, is an essential cellular immortalizing factor whose activation is a critical step in the progression to malignancy . An important agent maintaining the balance between proliferation, differentiation and apoptosis of intestinal epithelial cells in crypts is butyrate, which is formed in the gastrointestinal tract by anaerobic bacterial fermentation . It inhibits cell growth, induces differentiation and triggers apoptosis in neoplastic colonocytes . MATERIALS AND METHODS: In this study, the responses of adenocarcinoma (HT-29) and fetal (FHC) human colon cells to 5 mM sodium butyrate (NaBt) have been compared . RESULTS: Despite the similar general response of both cell lines to NaBt, i.e., G0/G1 arrest, decrease of growth rate and increase of differentiation (as indicated by alkaline phosphatase activity), they differ in the level and dynamics of the measured parameters . Telomerase activity and the level of mRNA for its catalytic subunit (hTERT) decline significantly after 48 hours, reaching a complete inhibition after 144 hours . While both cell lines show similar kinetics of hTERT transcriptional silencing, the down-regulation of telomerase activity is faster in FHC cells . Correspondingly, we show that a candidate posttranscriptional regulation step, differential splicing of hTERT mRNA, may be involved in the faster loss of telomerase activity in FHC cells . CONCLUSION: Differences in hTERT mRNA splicing may represent a useful marker of telomere metabolism in normal and malignant colon cells and that these changes may be connected with different cytokinetic patterns of these cells.

J Biol Chem, 2003 Sep 12, 278(37), 35552 - 7 Epub 2003 Jun 20.
Enhancement and selective production of phoslactomycin B, a protein phosphatase IIa inhibitor, through identification and engineering of the corresponding biosynthetic gene cluster; Palaniappan N et al.; Phoslactomycins (PLMs), potent and selective inhibitors of serine threonine phosphatases, are of interest for their antitumor and antiviral activity . Multiple analogs and low titers in the fermentation process have hampered the development of this class of natural products . The entire 75-kb PLM biosynthetic gene cluster of Streptomyces sp . HK-803 was cloned, sequenced, and analyzed . The loading domain and seven extension modules of the PLM polyketide synthase generate an unusual linear unsaturated polyketide chain containing both E- and Z-double bonds from a cyclohexanecarboxylic acid (CHC) primer . Hydroxylation of the CHC-derived side chain of the resulting PLM-B by PlmS2, and a subsequent esterification, produces the remaining PLM analogs . A new PCR targeting technology allowed rapid and facile allelic replacement of plmS2 . The resulting mutant selectively produced the PLM-B, at 6-fold higher titers than the wild type strain . This mutant and the biosynthetic gene cluster will facilitate engineered microbial production of hybrid PLMs with improved properties.

Bioorg Chem, 2003 Jun, 31(3), 227 - 36
Comparison of isotopic fractionation in lactic acid and ethanol fermentations; Zhang BL et al.; Pure D(-) and L(+) enantiomers of lactic acid were prepared by fermentation reactions with specific bacteria . In addition, naturally deuterated ethanol was prepared and converted into diastereoisomers using mandelic acid . Various sugars and nutrients were fermented into lactic acid in water having different deuterium contents and ethanol samples were obtained from yeast fermentation of sugars from different botanical origins . The methine and methylene groups in lactic acid and ethanol respectively show similar deuterium contents which are related to that found in the fermentation water . However, the methyl groups of both molecules are significantly different whatever the botanical origin of the carbon source in the fermentation medium.

J Antibiot (Tokyo), 2003 Apr, 56(4), 364 - 71
Ripromycin and other polycyclic macrolactams from Streptomyces sp . Tü 6239: taxonomy, fermentation, isolation and biological properties; Bertasso M et al.; Strain Tu 6239 was isolated from a soil sample collected in Brazil and determined as a new species of the genus Streptomyces . In the course of our HPLC-diode array screening program three metabolites were detected in the culture filtrate and mycelium extracts of strain Tu 6239 . They were characterised as members of the macrolactam group, the new compound ripromycin (1), the previously described ikarugamycin (2) and a new derivative of it, ikarugamycin epoxide (3) . They show antibiotic activities against gram-positive bacteria and cytostatic effects to various human tumor cell lines.

J Antibiot (Tokyo), 2003 Apr, 56(4), 344 - 50
Terreulactones A, B, C, and D: novel acetylcholinesterase inhibitors produced by Aspergillus terreus . I . Taxonomy, fermentation, isolation and biological activities; Cho KM et al.; In the course of screening for selective inhibitors of acetylcholinesterase from the microbial metabolites, four new meroterpenoid compounds, terreulactones A, B, C and D were isolated from solid state fermentation of Aspergillus terreus Fb000501 . They showed potent inhibitory activities against acetylcholinesterase with IC50 values in range of 0.06 to approximately 0.42 microM . In addition, they exhibited more than 500 to approximately 3000 times selectivity for acetylcholinesterase compared with butyrylcholinesterase.

J Antibiot (Tokyo), 2003 Apr, 56(4), 337 - 43
S14-95, a novel inhibitor of the JAK/STAT pathway from a Penicillium species; Erkel G et al.; In a search for new inhibitors of the IFN-gamma mediated signal transduction in HeLa S3 cells using secreted alkaline phosphatase (SEAP) as reporter gene, a novel compound, designated as S14-95 was isolated from fermentations of the imperfect fungus Penicillium sp . 14-95 . The compound inhibits the IFN-gamma mediated expression of the reporter gene with IC50 values of 2.5 to approximately 5 microg/ml (5.4 to approximately 10.8 microM) . Furthermore the compound inhibited the expression of the proinflammatory enzymes COX-2 and NOS II at 5 microg/ml (10.8 microM) in LPS/IFN-gamma stimulated J774 mouse macrophages . Studies on the mode of action of the compound revealed that the inhibition of the IFN-gamma dependent signaling pathway is caused by an inhibition of the phosphorylation of the STAT1alpha transcription factor . In addition, S14-95 inhibited the activation of the p38 MAP kinase, which is involved in the inducible expression of many proinflammatory genes.

J Anim Sci, 2003 Jun, 81(6), 1611 - 27
Biohydrogenation of unsaturated fatty acids in continuous culture fermenters during digestion of orchardgrass or red clover with three levels of ground corn supplementation; Loor JJ et al.; Diet digestibility and outputs of biohydrogenation intermediates were assessed in a continuous culture of ruminal microorganisms . Orchardgrass or red clover harvested and frozen during spring or fall served as the primary substrates for fermentation . During 10-d incubations, fermenters were fed thawed forage (50 g of DM/d), forage (42 g/d) plus 8 g/d of corn, or forage (34 g/d) plus 16 g/d of corn . Effluents from the last 3 d of incubation were composited for analyses . Starch input increased from 5 to 27% of DM as corn input increased from 0 to 16 g/d . Corn input reduced (P < 0.01) pH, increased (P < 0.01) microbial DM yield, and increased (P = 0.01) digestibility of DM, NDF, CP, and nonstructural carbohydrates . Overall, apparent hydrogenation (percentage) of cis9-18:1, 18:2n-6, and 18:3n-3 was greater (P < 0.05) with orchardgrass than clover . Hydrogenation of cis9-18:1 and 18:2n-6 increased (P = 0.01), but hydrogenation of 18:3n-3 decreased (P = 0.01) linearly due to corn input, regardless of forage . As a result, output of trans11, cis15-18:2 also decreased (P = 0.01) . Average output of cis9,trans11-18:2 was greater (P = 0.01) for clover (1.3 mg/d) compared with orchardgrass (0.6 mg/d), but corn input with either forage increased (P = 0.01) cis9,trans11-18:2 output by 205% . Output of trans11-18:1 was greater (P = 0.01) from orchardgrass compared with clover (174 vs . 90 mg/d), but corn increased (P = 0.01) trans11-18:1 output only from clover fermentations . Output of trans10-18:1 was greater (P = 0.01) in response to orchardgrass compared with clover (10 vs . 4 mg/d), but corn addition doubled the output regardless of forage type . Output of trans10,cis12-18:2, which did not differ due to forage type, increased (P = 0.01) twofold in response to corn . Cis9,cis11-18:2 was a primary conjugated isomer produced from forage fermentations, but its output decreased (P = 0.03) in response to corn input . When inputs of 18:2n-6 plus 18:3n-3 were less than 0.9% of total DM (clover), hydrogenation was low (87%) . When 18:2n-6 plus 18:3n-3 inputs were from 1.2 to 1.5% of total DM (orchardgrass), hydrogenation averaged 96% . Despite greater hydrogenation, incremental additions of cis9-18:1 and 18:2n-6 from corn grain increased (P < 0.05) outputs of trans10-18:1, trans11-18:1, trans10,cis12-18:2, cis9,trans11-18:2, and trans,trans-18:2 in effluent . Results suggest that forage species alone or in combination with corn grain can alter hydrogenation and profiles of intermediates to varying degrees.

Yi Chuan Xue Bao, 2003 Apr, 30(4), 370 - 5
{Effect of growth, temperature and media on the expression of secE promoter of Streptomyces lividans TK24}; Wang LF et al.; Streptomyces are Gram-positive, filamentous soil bacteria, which produce a wide variety of metabolites that are currently being exploited in both medicine and agriculture . Moreover, Streptomyces lividans is used as a host strain to effectively express and secrete heterologous proteins to culture media . Genes encoding Sec proteins responsible for the translocation of preproteins have been identified in S . lividans . SecYEG, a complex of integral membrane proteins SecY, SecE, and SecG in the cytoplasmic membrane, constitutes a pathway for polypeptide movement . SecA plays a central role in translocation as it is the site of preprotein entry into the translocase and it is the only ATPase essential for preprotein translocation . The role of SecD and SecF is to regulate the movement of the translocating protein . A better understanding of their regulatory mechanism could help to develop S . lividans strains with hyper-secretory capacity . In this study, a reporter system was used to investigate the regulatory mechanism of secE promoter . Upstream sequence (496 bp) of secE of S . lividans TK24 was cloned and characterized in a promoter probe vector pIJ4083 upstream of promoterless xylE . Sequencing analysis revealed that secE upstream sequence of S . lividans shares 99.8% homology with the secE promoter of S . coelicolor . The transcriptive activity of this fragment approximates vsi promoter in CMI medium, a strong promoter from S . venezuelae . The activity of secE promoter was observed in different growth phase, culture temperature and culture media . The expression level of secE was higher during the log phase, but decreased at the beginning of the stationary phase . At growth temperature of 28 degrees C, the expression level was much higher than that at 37 degrees C . When the bacteria were incubated in NB, Phage, and CM medium respectively, the expression level of secE promoter was different at a certain fermentation time, but all reached the highest level at log phase . Further analysis revealed that glucose may repress secE promoter expression.

Arch Microbiol, 2003 Aug, 180(2), 134 - 41 Epub 2003 Jun 13.
Xylose metabolism in the anaerobic fungus Piromyces sp . strain E2 follows the bacterial pathway; Harhangi HR et al.; The anaerobic fungus Piromyces sp . strain E2 metabolizes xylose via xylose isomerase and d-xylulokinase as was shown by enzymatic and molecular analyses . This resembles the situation in bacteria . The clones encoding the two enzymes were obtained from a cDNA library . The xylose isomerase gene sequence is the first gene of this type reported for a fungus . Northern blot analysis revealed a correlation between mRNA and enzyme activity levels on different growth substrates . Furthermore, the molecular mass calculated from the gene sequence was confirmed by gel permeation chromatography of crude extracts followed by activity measurements . Deduced amino acid sequences of both genes were used for phylogenetic analysis . The xylose isomerases can be divided into two distinct clusters . The Piromyces sp . strain E2 enzyme falls into the cluster comprising plant enzymes and enzymes from bacteria with a low G+C content in their DNA . The d-xylulokinase of Piromyces sp . strain E2 clusters with the bacterial d-xylulokinases . The xylose isomerase gene was expressed in the yeast Saccharomyces cerevisiae, resulting in a low activity (25+/-13 nmol min(-1)mg protein(-1)) . These two fungal genes may be applicable to metabolic engineering of Saccharomyces cerevisiae for the alcoholic fermentation of hemicellulosic materials.

Asia Pac J Clin Nutr, 2003, 12(2), 129 - 37
Captain Cook's beer: the antiscorbutic use of malt and beer in late 18th century sea voyages; Stubbs BJ; The custom of allowing British seamen the regular use of fermented liquor is an old one . Ale was a standard article of the sea ration as early as the fourteenth century . By the late eighteenth century, beer was considered to be at once a food (a staple beverage and essential part of the sea diet), a luxury (helping to ameliorate the hardship and irregularity of sea life) and a medicine (conducive to health at sea) . In particular, beer and its precursors, wort and malt, were administered with the aim of preventing and curing scurvy . This paper examines the use of malt and beer during late eighteenth century British sea voyages, particularly their use as antiscorbutic agents, focusing on James Cook's three voyages during the period 1768-1780 . Cook administered sweet wort (an infusion of malt), beer (prepared from an experimental, concentrated malt extract), and spruce beer (prepared mainly from molasses), among many other items, in his attempts to prevent and to cure scurvy . Despite the inconclusive nature of his own experiments, he reported favourably after his second voyage (1772-1775) on the use of wort as an antiscorbutic sea medicine (for which purpose it is now known to be useless) . Cook thereby lent credibility to erroneous medical theories about scurvy, helping to perpetuate the use of ineffective treatments and to delay the discovery of a cure for the disorder.

Int J Food Microbiol, 2003 Aug 15, 85(1-2), 127 - 36
Effects of fermentation temperature and Saccharomyces species on the cell fatty acid composition and presence of volatile compounds in wine; Torija MJ et al.; Low temperature alcoholic fermentations are becoming more frequent due to the wish to produce wines with more pronounced aromatic profiles . However, their biggest drawback is the high risk of stuck and sluggish fermentations . Changes in the plasma membrane composition may be an adaptive response to low temperature fermentations . The production of volatile compounds and the changes in the membrane fatty acids were determined by GC to show the degree of cell adaptation and performance at low temperatures (13 degrees C) taking 25 degrees C as reference . The tests were done in two strains of Saccharomyces cerevisiae and one strain of Saccharomyces bayanus . Low temperatures restricted yeast growth and lengthened the fermentations . The analysis of plasma membrane fatty acids showed that dry yeasts had similar levels of unsaturation, between 70% and 80%, with no medium-chain fatty acids (MCFA) . Long-chain saturated fatty acids (SFA) were the most frequent membrane fatty acids throughout the fermentations . Lipid composition changed with the growth temperature . The optimal membrane fluidity at low temperatures was modulated by changes in the unsaturation degree in S . cerevisiae strains . In S . bayanus, however, this change in the unsaturated fatty acid (UFA) percentage was not observed at different growth temperatures but the concentration of MCFA at low fermentation temperatures was higher . Concentrations of volatile compounds were higher in wines produced at lower temperatures and depended on the strain.

Int J Food Microbiol, 2003 Aug 15, 85(1-2), 111 - 25
Changes in selected biochemical and sensory parameters as affected by the superficial inoculation of Penicillium camemberti on dry fermented sausages; Bruna JM et al.; An atoxigenic strain of Penicillium camemberti was superficially inoculated on fermented sausages in an attempt to improve their sensory properties . The growth of this mould on the surface of the sausages resulted in an intense proteolysis and lipolysis, which caused an increase in the concentration of free amino acids, free fatty acids (FFA) and volatile compounds . Many of these were derived from amino acid catabolism and were responsible for the "ripened flavour", i.e . branched aldehydes and the corresponding alcohols, acids and esters . The development of the fungal mycelia on the surface of the sausages also protected lipids from oxidation, resulting in both lower 2-thiobarbituric acid (TBARS) values and lipid oxidation-derived compounds, such as aliphatic aldehydes and alcohols . The sensory analysis of superficially inoculated sausages showed clear improvements in odour and flavour and, as a consequence, in the overall quality of the sausages . Therefore, this strain is proposed as a potential starter culture for dry fermented sausage production.

Circ J, 2003 Jun, 67(6), 530 - 4
Feasibility and effect on blood pressure of 6-week trial of low sodium soy sauce and miso (fermented soybean paste); Nakamura M et al.; A double-blind, randomized placebo-controlled study was conducted to evaluate the feasibility of the long-term use of low-sodium soy sauce and miso in the general Japanese population and its effect on blood pressure (BP) . Forty men and 24 women were randomly allocated to a low-sodium group (n=32) or a control group (n=32) . Low-sodium soy sauce and miso, which were approximately 25% and 20% lower in salt content than common soy sauce and miso, were used in the study . The change in BP after a 6-week intervention was evaluated . There were no significant differences in age, sex, body mass index, BP or hypertension between the 2 groups before intervention . After the 6-week intervention, no significant change in BP was observed in the entire cohort . However, in those aged 40 years and older, 6.4 mmHg net reduction in diastolic BP with no significant change in systolic BP was noted in the low-sodium group . Taste evaluation for the low-sodium seasoning was considerably good . Replacing soy sauce and miso of the common type with the low-sodium alternative is feasible in the general population and could be the basis for a salt reduction strategy in the Japanese diet.

Int J Syst Evol Microbiol, 2003 May, 53(Pt 3), 815 - 21
Spirochaeta americana sp . nov., a new haloalkaliphilic, obligately anaerobic spirochaete isolated from soda Mono Lake in California; Hoover RB et al.; A novel, obligately anaerobic, mesophilic, haloalkaliphilic spirochaete, strain ASpG1(T), was isolated from sediments of the alkaline, hypersaline Mono Lake in California, USA . Cells of the Gram-negative strain were motile and spirochaete-shaped with sizes of 0.2-0.22 x 8-18 microm . Growth of the strain was observed between 10 and 44 degrees C (optimum 37 degrees C), in 2-12% (w/v) NaCl (optimum 3% NaCl) and between pH 8 and 10.5 (optimum pH 9.5) . The novel strain was strictly alkaliphilic, required high concentrations of carbonates in the medium and was capable of utilizing D-glucose, fructose, maltose, sucrose, starch and D-mannitol . End products of glucose fermentation were H2, acetate, ethanol and formate . Strain ASpG(T) was resistant to kanamycin and rifampicin, but sensitive to gentamicin, tetracycline and chloramphenicol . The G + C content of its DNA was 58.5 mol% . DNA-DNA hybridization analysis of strain ASpG1(T) with its most closely related species, Spirochaeta alkalica Z-7491(T), revealed a hybridization value of only 48.7% . On the basis of its physiological and molecular properties, strain ASpG1(T) appears to represent a novel species of the genus Spirochaeta, for which the name Spirochaeta americana is proposed (type strain ASpG1(T) =ATCC BAA-392(T) = DSM 14872(T)).

J Plant Physiol, 2003 May, 160(5), 565 - 8
Anoxia tolerance in rice roots acclimated by several different periods of hypoxia; Kato-Noguchi H; Rice (Oryza sativa L.) seedlings were subjected to hypoxic pretreatment (H-PT; incubated in 5% O2 atmosphere) for various lengths of time followed by an anoxic stress . Anoxia tolerance of rice roots was improved with increasing duration of H-PT, but longer H-PT than 12 h gave no additional improvement . Concentrations of ATP and ethanol, and activities of pyruvate decarboxylase (EC 4.1.1.1) and alcohol dehydrogenase (EC 1.1.1.1) in the roots were increased by H-PT, and the times and patterns of increasing in these concentrations and activities were similar to those of increasing in the anoxia tolerance . These results suggest that the H-PT may increase anoxia tolerance due to maintenance of ATP levels with rapid induction of ethanolic fermentation, and hypoxic acclimation may occur within 12 h.

Plant Physiol, 2003 Jun, 132(2), 968 - 78 Epub 2003 Apr 24.
The pyruvate decarboxylase1 gene of Arabidopsis is required during anoxia but not other environmental stresses; Kursteiner O et al.; Ethanolic fermentation is classically associated with flooding tolerance when plant cells switch from respiration to anaerobic fermentation . However, recent studies have suggested that fermentation also has important functions in the presence of oxygen, mainly in germinating pollen and during abiotic stress . Pyruvate decarboxylase (PDC), which catalyzes the first step in this pathway, is thought to be the main regulatory enzyme . Here, we characterize the PDC gene family in Arabidopsis . PDC is encoded by four closely related genes . By using real-time quantitative polymerase chain reaction, we determined the expression levels of each individual gene in different tissues, under normal growth conditions, and when the plants were subjected to anoxia or other environmental stress conditions . We show that PDC1 is the only gene induced under oxygen limitation among the PDC1 gene family and that a pdc1 null mutant is comprised in anoxia tolerance but not other environmental stresses . We also characterize the expression of the aldehyde dehydrogenase (ALDH) gene family . None of the three genes is induced by anoxia but ALDH2B7 reacts strongly to ABA application and dehydration, suggesting that ALDH may play a role in aerobic detoxification of acetaldehyde . We discuss the possible role of ethanolic fermentation as a robust back-up energy production pathway under adverse conditions when mitochondrial function is disturbed.

Arch Tierernahr, 2003 Feb, 57(1), 65 - 81
Effects of Moringa oleifera seed extract on rumen fermentation in vitro; Hoffmann EM et al.; Moringa oleifera is a pantropical tree of the family Moringaceae . A previously undescribed property of an aqueous extract from the seeds of this plant is the modulation of ruminal fermentation patterns, especially protein degradation, as demonstrated in a short-term batch incubation system . Gas, short chain fatty acids (SCFA) and cellulolytic enzyme activities were determined as general fermentation parameters . A dot blot assay able to directly detect true protein in rumen fluid samples was used to quantify protein degradation . For complex substrates the interpretation of protein degradation profiles was amended by polyacrylamide gel electrophoresis (PAGE) of the samples . When incubated with pure carbohydrates at a concentration of 1 mg ml(-1), the extract reduced microbial degradation of the model protein, bovine serum albumin (BSA), such that its concentration was at least 40% above the control after 12 h of incubation . Total protein degradation was thus delayed by approximately 9 h . When fermented along with wheat straw, leaf protein (Rubisco) was almost entirely protected during 12 h of fermentation . The degradation of soy proteins was retarded by at least 4-6 h, depending on the protein band . There were strong side effects on the fermentation of pure cellulose (SCFA yield-60% after 12 h), whereas cellobiose and starch fermentation were less affected (-18 and -8%, respectively) . When the complex substrates were fermented, SCFA yield was reduced by approximately 30% after 12 h . In our work we clearly demonstrate the efficacy of the new substance, which is neither a tannin nor a saponin, in an in vitro system, using pure as well as complex substrates . The properties shown in vitro for the crude extract suggest that it could have a positive effect on the protein metabolism of ruminants under intensive management and that negative side effects can be overcome by an optimized dosage . If the chemical nature of the active substance and its mechanism of action can be clarified, it may provide an alternative to replace critical synthetic feed additives (such as antibiotics) for high yielding dairy cows.

J Chromatogr A, 2003 May 2, 995(1-2), 161 - 9
Different multidimensional chromatographic approaches applied to the study of wine malolactic fermentation; Fernandes L et al.; Different multidimensional chromatographic techniques were used to study wine aroma pattern changes during malolactic fermentation (MLF) . Ethyl lactate enantiomeric ratios were determined using on-line multidimensional gas chromatography . The values found agree with a spontaneous MLF . Off-line multidimensional HPLC/GC was used to deconvolute and enrich the sample and ease enantioselective chromatography . Chiral compound enantiomeric ratio changes during MLF were monitored . Evaluation of enantiomeric ratio changes during MLF has never been studied . (R,R), (S,S) and meso-butane-2,3-diol and pentane-2,4-diol (reported in wines for the first time) were submitted to untrained sensory panel tests . All stereoisomers revealed different sensory notes; pentane-2,4-diol showed an aromatic impact.

Huan Jing Ke Xue, 2003 Mar, 24(2), 54 - 9
{Cooperation of mixed culturing bacteria in the hydrogen production by fermentation}; Lin M et al.; In order to discuss the cooperation of H2-producing fermentation bacteria (HPFB) in mixed culture, a batch test fed with glucose and complex organic substance (starch, beef, PEP and peptone) respectively, was conducted to investigate the effects of mixed culture on H2-producing ability . For the systemic and accurate analysis, three kinds of the mixed culture were use to this batch test . It included that the mixed culture with five strains of HPFB (B49, H1, LM12, LM11 and B51), B49 and three stains of non-HPFB (L10, Bacteroide 3-2, Sporobacterl), and B49 and hydrogen producing sludge . The results showed that the cooperation of mixed culturing bacteria was conditional on the substrates . When fed with glucose, which were easily utilized by HPB, the H2-producing ability of HPB was restrained because of the competition on the co-substrate between HPB and other fermentation bacteria, and it was quite difficult for the cooperation of mixed culturing bacteria to be performed . When fed with complex organic substance, the H2-producing ability of HPB was enhanced via the cooperation of mixed culturing bacteria . Furthermore, a strategy was put forward, that is, different kind of HPB cultures could be adopted according to the difference of substrates.

Biotechnol Bioeng, 2003 Aug 20, 83(4), 395 - 9
Critical evaluation of sampling techniques for residual glucose determination in carbon-limited chemostat culture of Saccharomyces cerevisiae; Mashego MR et al.; In this paper, three sampling techniques for rapid quenching of cellular metabolism and subsequent separation of cells from fermentation broth are compared: (i) quick freezing of fermentation broth directly in liquid nitrogen; (ii) quenching metabolism by exposing the fermentation broth to stainless steel beads (4-mm diameter) in a filter syringe precooled to -18 degrees C; and (iii) withdrawal of the filtrate through a 0.45 microm filter attached to a syringe and a needle inserted directly into the fermentor . It was concluded that use of liquid nitrogen as a quenching method to rapidly arrest cellular metabolism, for quantitative analysis of extracellular glucose, is not a very reliable method and that the filter syringe steel beads work very well .

Biotechnol Bioeng, 2003 Aug 20, 83(4), 376 - 85
Novel type of in situ extraction: Use of solvent containing microcapsules for the bioconversion of 2-phenylethanol from L-phenylalanine by Saccharomyces cerevisiae; Stark D et al.; A novel in situ product removal (ISPR) method that uses microcapsules to extract inhibitory products from the reaction suspension is introduced into fermentation technology . More specifically, L-phenylalanine (L-Phe) was transformed by Saccharomyces cerevisiae to 2-phenylethanol (PEA), which is inhibitory toward the yeast . In order to continuously remove PEA from the vicinity of the cells, the reaction suspension was brought into contact with capsules of 2.2-mm diameter that had a hydrophobic core of dibutyl sebacate and an alginate-based wall . This novel process combines the advantages of a normal in situ extraction process (fast mass transfer and simple process set-up) with the benefits of a membrane-based process (reduction of the solvent toxicity and avoidance of stable emulsions) . In particular, the microbial cells are shielded from the phase toxicity of the organic solvent by a hydrogel layer surrounding the organic core . By placing the microcapsules into the fermenter, the final overall concentration of PEA in a fed-batch culture was increased from 3.8 to 5.6 g/L because a part of the inhibitory product dissolved in the dibutyl sebacate core . In another fermentation experiment, the capsules were placed in a fluidized bed that was connected via a loop to the fermenter . In addition, the fluidized bed was connected via a second loop to a back-extractor to regenerate the capsules . By alternating the extraction and back-extraction cycles, it was possible to limit the PEA concentration of the fed-batch culture in the fermenter to 2.4 g/L while producing important quantities of PEA that accumulated in an external reservoir .

J Biol Chem, 2003 Aug 22, 278(34), 31774 - 80 Epub 2003 Jun 10.
Mammalian mitochondrial initiation factor 2 supports yeast mitochondrial translation without formylated initiator tRNA; Tibbetts AS et al.; Initiation of protein synthesis in mitochondria and chloroplasts is widely believed to require a formylated initiator methionyl-tRNA (fMet-tRNAfMet) in a process involving initiation factor 2 (IF2) . However, yeast strains disrupted at the FMT1 locus, encoding mitochondrial methionyl-tRNA formyltransferase, lack detectable fMet-tRNAfMet but exhibit normal mitochondrial function as evidenced by normal growth on non-fermentable carbon sources . Here we show that mitochondrial translation products in Saccharomyces cerevisiae were synthesized in the absence of formylated initiator tRNA . ifm1 mutants, lacking the mitochondrial initiation factor 2 (mIF2), are unable to respire, indicative of defective mitochondrial protein synthesis, but their respiratory defect could be complemented by plasmid-borne copies of either the yeast IFM1 gene or a cDNA encoding bovine mIF2 . Moreover, the bovine mIF2 sustained normal respiration in ifm1 fmt1 double mutants . Bovine mIF2 supported the same pattern of mitochondrial translation products as yeast mIF2, and the pattern did not change in cells lacking formylated Met-tRNAfMet . Mutant yeast lacking any mIF2 retained the ability to synthesize low levels of a subset of mitochondrially encoded proteins . The ifm1 null mutant was used to analyze the domain structure of yeast mIF2 . Contrary to a previous report, the C terminus of yeast mIF2 is required for its function in vivo, whereas the N-terminal domain could be deleted . Our results indicate that formylation of initiator methionyl-tRNA is not required for mitochondrial protein synthesis . The ability of bovine mIF2 to support mitochondrial translation in the yeast fmt1 mutant suggests that this phenomenon may extend to mammalian mitochondria as well.

Fertil Steril, 2003 Jun, 79(6), 1434 - 8
Hyaluronic acid can successfully replace albumin as the sole macromolecule in a human embryo transfer medium; Simon A et al.; OBJECTIVE: To examine the effect on pregnancy and implantation rates when highly purified, fermentation-based hyaluronic acid was the only macromolecule supplement to the transfer medium in a human IVF program . DESIGN: Prospective randomized study . SETTING: In vitro fertilization center in an academic medical institution . PATIENT(S): Eighty patients were included in this prospective randomized double blind study . Inclusion criteria were age </=35 years, the availability of at least three embryos eligible for transfer on day 3 after fertilization, and no more than three previous embryo transfer attempts . INTERVENTION(S): All embryos were cultured in P1 medium containing 10% synthetic serum substitute (SSS) until day 3 . Patients were randomly allocated to two groups; in treatment group A (40 patients), embryos were transferred to P1 medium supplemented with 0.5 mg/mL hyaluronic acid for 5-10 min before their intrauterine transfer . In the control group B (40 patients), embryos were transferred, as routinely performed, in P1 medium containing 10% SSS . MAIN OUTCOME MEASURES: Clinical pregnancy and implantation rates . RESULT(S): The mean age of the female partner was 28.7 +/- 3.3 years and 29.7 +/- 3.8 years for groups A and B, respectively . In group A, 103 embryos were transferred and in group B, 97 embryos were transferred for a similar mean number of 2.6 +/- 0.6 and 2.4 +/- 0.5 embryos/transfer, respectively . Twenty-five pregnancies were achieved in group A, and 21 pregnancies in group B . This led to a comparable clinical pregnancy and implantation rates of 62.5% and 34% as compared to 52% and 26.8% for groups A and B, respectively . CONCLUSION(S): Hyaluronic acid can successfully replace albumin as a sole macromolecule in a human embryo transfer medium and result in high pregnancy and implantation rates . The use of this supplement is an important step in the development of human embryo culture media free of blood-derived additives.

J Agric Food Chem, 2003 Jun 18, 51(13), 3874 - 9
Polyphenols from Honeybush tea (Cyclopia intermedia); Kamara BI et al.; The fermented leaves and stems of Cyclopia intermedia are used to brew Honeybush tea, a herbal tea indigenous to South Africa . The plant is also used to manufacture a sweet herbal infusion used for restorative properties such as soothing coughs and alleviating bronchial complaints including tuberculosis, pneumonia, and catarrh . It is claimed to have a low tannin content and no caffeine and contains various antioxidants . Continued investigations into the phenolic content of the leaves and stems of C . intermedia yielded tyrosol and a methoxy analogue, 2-{4-{O-alpha-apiofuranosyl-(1' '-->6')-beta-d-glucopyranosyloxy}phenyl}ethanol, 4-{O-alpha-apiofuranosyl-(1' '-->2')-beta-d-glucopyranosyloxy}benzaldehyde, five glycosylated flavonols, two isoflavones, four flavanones, two isoflavones, and two flavones . Structure elucidation was done by NMR, CD, and MS methods . Because flavonoids are presumed to contribute significantly toward the scavenging effects of active oxygen species, our results indicate that the tentative claimed health-promoting properties may be attributed to the presence of these and other phenolics in C . intermedia.

Ann N Y Acad Sci, 2003 May, 988, 269 - 81
Chemical versus biological explanations: interdisciplinarity and reductionism in the 19th century life sciences; Schummer J; This paper analyzes four controversies in the 19th century life sciences: the nature of fermentation, the nature of infectious diseases, the generation of life from inanimate matter, and vitalism . All these controversies appear to concern chemical versus biological explanations, suggesting that reduction of biology to chemistry was the common underlying issue . My analysis rejects such interpretations, including the labels for explanation, and instead points out sophisticated forms of interdisciplinarity between chemistry, medicine, and biology in the first three debates . I argue that the philosophically favored perspective on reductionism, historically induced by a few physicians in the fourth debate, leads us astray from understanding interdisciplinary research.

Public Health Nutr, 2003 Jun, 6(4), 333 - 40
Yoghurt in the Spanish diet: nutritional implications and socio-cultural aspects of its consumption; Capdevila F et al.; OBJECTIVES: This study aims to analyse the differences, with regard to socio-cultural characteristics and dietary habits, between low and high consumers of yoghurt and other fermented dairy product desserts, and the nutritional significance of these differences . METHODS: We analysed the diet of a healthy population (4-65 years), using the 24-hour recall method, on three non-consecutive days . The participants were grouped by age and sex and were also divided into tertiles on the basis of yoghurt consumption . We compared energy and nutrient intakes, educational level and socio-economic status in the low consumption (LC) group and the high consumption (HC) group . RESULTS: In general there were no significant differences in energy intake or nutritional profile between LC and HC groups . The only significant difference was in the percentage of energy provided by lipids, which was significantly lower in HC women, possibly due to the high number of women in this group who consumed low-fat yoghurt . There were significant differences in the distribution of HC and LC subjects according to the three educational levels but not according to socio-economic status . CONCLUSION: The fact of being a high consumer of fermented dairy products took place in the framework of other dietary changes that compensated for this high consumption, resulting in the absence of significant differences in energy intake and nutritional profile between HC and LC subjects . The only exception was found in women who consumed low-fat dairy products . There was a relationship between high consumption of fermented dairy products and educational level.

Yeast, 2003 Jun, 20(8), 747 - 59
Response to acetaldehyde stress in the yeast Saccharomyces cerevisiae involves a strain-dependent regulation of several ALD genes and is mediated by the general stress response pathway; Aranda A et al.; One of the stress conditions that yeast may encounter is the presence of acetaldehyde . In a previous study we identified that, in response to this stress, several HSP genes are induced that are also involved in the response to other forms of stress . Aldehyde dehydrogenases (ALDH) play an important role in yeast acetaldehyde metabolism (e.g . when cells are growing in ethanol) . In this work we analyse the expression of the genes encoding these enzymes (ALD) and also the corresponding enzymatic activities under several growth conditions . We investigate three kinds of yeast strains: laboratory strains, strains involved in the alcoholic fermentation stage of wine production and flor yeasts (responsible for the biological ageing of sherry wines) . The latter are very important to consider because they grow in media containing high ethanol concentrations, and produce important amounts of acetaldehyde . Under several growth conditions, further addition of acetaldehyde or ethanol in flor yeasts induced the expression of some ALD genes and led to an increase in ALDH activity . This result is consistent with their need to obtain energy from ethanol during biological ageing processes . Our data also suggest that post-transcriptional and/or post-translational mechanisms are involved in regulating the activity of these enzymes . Finally, analyses indicate that the Msn2/4p and Hsf1p transcription factors are necessary for HSP26, ALD2/3 and ALD4 gene expression under acetaldehyde stress, while PKA represses the expression of these genes .

Appl Biochem Biotechnol, 2003 Apr-Jun, 109(1-3), 207 - 25
Reductive decolorization of a textile reactive dyebath under methanogenic conditions; Fontenot EJ et al.; The objective of the present study was to assess the biological decolorization of an industrial, spent reactive dyebath and its three dye components (Reactive Blue 19 {RB 19}, Reactive Blue 21 {RB 21}, and Reactive Red 198 {RR 198}) under methanogenic conditions . Using a mixed, methanogenic culture, batch assays were performed to evaluate the rate and extent of color removal as well as any potential toxic effects . Overall, a high rate and extent of color removal (>10 mg/{L.h} and 88%, respectively) were observed in cultures amended with either RB 19 (an anthraquinone dye) or spent dyebath at an initial dye concentration of 300 mg/L (expressed as RB 19 equivalent) and 30 g/L of NaCl . Inhibition of acidogenesis and, to a larger degree, of methanogenesis resulting in accumulation of volatile fatty acids was observed in both RB 19- and spent dyebath-amended cultures . RB 21 (a phthalocyanine dye) and RR 198 (an azo dye) tested at an initial concentration of 300 mg/L did not result in any significant inhibition of the mixed methanogenic culture . Based on results obtained with cultures amended with RB 19 with and without NaCl, as well as a control culture amended with 30 g/L of NaCl, salt was less inhibitory than either RB 19 or the dyebath . Therefore, the toxic effect of the spent dyebath is at least partially attributed to its major dye component RB 19 and NaCl . Further testing of the effect of RB 19 decolorization products on the methanogenic activity in the absence of NaCl demonstrated that these products are much less inhibitory than the parent dye . Although color removal occurred despite the severe culture inhibition, biological decolorization of full-strength reactive spent dyebaths using methanogenic cultures in a repetitive, closed-loop system is not deemed feasible . For this reason, a fermentative and halotolerant culture was developed and successfully used in our laboratory for the decolorization of industrial reactive dyebaths with 100 g/L of NaCl.

Oncol Rep, 2003 Jul-Aug, 10(4), 875 - 80
Preventive effect of fermented brown rice and rice bran on diethylnitrosoamine and phenobarbital-induced hepatocarcinogenesis in male F344 rats; Katayama M et al.; Epidemiological and preclinical studies have suggested that nutrition plays an important role in the etiology of cancer . Our group previously demonstrated that rice germ or fermented brown rice has a preventive effect on colorectal carcinogenesis . The experiment described here was examined for the potential anticancer properties of brown rice fermented by Aspergillus Oryzae (FBRA) in male F344 rats using inhibition of diethylnitrosoamine (DEN) and phenobarbital (PB)-induced hepatocarcinogenesis as the measure of preventive efficacy when this agent was administered at 5% and 10% levels in diet during initiation phase (during and until 1 week after carcinogen treatment) or post-initiation phase (beginning 1 week after carcinogen treatment) of the carcinogenesis . Rats were sacrificed 20 weeks after the initiation of DEN treatment (200 mg/kg body weight, once weekly for 3 weeks) . Expression of liver tumors was evaluated histopathologically . Administration of 10% FBRA in the diet during the initiation phase significantly decreased the incidence (43% vs . 8%) and multiplicity (0.5+/-0.6 vs . 0.1+/-0.3) of hepatocellular carcinoma (HCC) as compared to those given the control diet . In addition, 5% and 10% of FBRA in the diet during post-initiation phase significantly decreased the incidence of HCC (43% vs . 15% and 9%, respectively) and multiplicity of hepatocellular adenoma (4.7+/-3.7 vs . 2.1+/-2.2 and 2.4+/-1.4, respectively) and HCC (0.5+/-0.6 vs . 0.2+/-0.4 and 0.1+/-0.3, respectively) . These data prove that FBRA has an inhibitory effect on the hepatocarcinogenesis in rats . FBRA could be a promising chemopreventive agent for human liver as well as colorectal neoplasia.

J Biol Chem, 2003 Aug 22, 278(34), 32141 - 9 Epub 2003 Jun 05.
Transcriptional, proteomic, and metabolic responses to lithium in galactose-grown yeast cells; Bro C et al.; Lithium is highly toxic to yeast when grown in galactose medium mainly because phosphoglucomutase, a key enzyme of galactose metabolism, is inhibited . We studied the global protein and gene expression profiles of Saccharomyces cerevisiae grown in galactose in different time intervals after addition of lithium . These results were related to physiological studies where both secreted and intracellular metabolites were determined . Microarray analysis showed that 664 open reading frames were down-regulated and 725 up-regulated in response to addition of lithium . Genes involved in transcription, translation, and nucleotide metabolism were down-regulated at the transcriptional level, whereas genes responsive to different stresses as well as genes from energy reserve metabolism and monosaccharide metabolism were up-regulated . Compared with the proteomic data, 26% of the down-regulated and 48% of the up-regulated proteins were also identified as being changed on the mRNA level . Functional clusters obtained from proteome data were coincident with transcriptional clusters . Physiological studies showed that acetate, glycerol, and glycogen accumulate in response to lithium, as reflected in expression data, whereas a change from respiro-fermentative to respiratory growth could not be predicted from the expression analyses.

Nutr Cancer, 2003, 45(1), 84 - 92
Butyrate restores motile function and actin cytoskeletal network integrity in apc mutated mouse colon epithelial cells; Forest V et al.; Loss of function of the Apc gene product is an early and frequent event in colorectal carcinogenesis . Altered migration of intestinal epithelial cells has been described in vivo in the Min mouse Apc+/Min model . Using cell lines established from this model we show in vitro that Apc+/Min cells are less motile than Apc+/+ cells and exhibit a disordered actin cytoskeletal network . This would increase the probabilities of the initiated cell to acquire additional genetic alterations leading to neoplasia . Butyrate, a product of indigestible carbohydrate fermentation by the colonic flora, is able to restore both motility and actin cytoskeletal organization . This feature may contribute to explain the protective effect exerted by butyrogenic diets on colon carcinogenesis in animal models.

Biotechnol Prog, 2003 May-Jun, 19(3), 1091 - 4
Pulsed feeding during fed-batch Aspergillus oryzae fermentation leads to improved oxygen mass transfer; Bhargava S et al.; Productivity in many fungal fermentations is detrimentally affected by high broth viscosity and consequent reduced oxygen mass transfer capacity . The goal here was to determine whether pulsed feeding of limiting carbon in a fungal fermentation could lead to reduced viscosity and improved oxygen mass transfer . As a model, an industrially relevant recombinant strain of Aspergillus oryzae was grown in carbon-limited, fed-batch mode . Maltodextrin was used as a carbon source and was added either continuously or in 1.5-min pulses, 3.5 min apart . In both feeding modes the same total amount of carbon was added, and carbon feed rate was at sufficiently low levels to ensure cultures were always carbon-limited . Compared to continuous feeding, pulsed addition of substrate led to smaller fungal elements, which resulted in a significant reduction in broth viscosity . This in turn led to higher dissolved oxygen concentrations and increased oxygen uptake rates during pulsed feeding.

Biotechnol Prog, 2003 May-Jun, 19(3), 1085 - 90
Study of protein splicing and intein-mediated peptide bond cleavage under high-cell-density conditions; Sharma S et al.; Protein splicing elements (inteins), capable of catalyzing controllable peptide bond cleavage reactions, have been used to separate recombinant proteins from affinity tags during affinity purification . Since the inteins eliminate the use of a protease in the recovery process, the intein-mediated purification system has the potential to significantly reduce recovery costs for the industrial production of recombinant proteins . Thus far, the intein system has only been examined and utilized for expression and purification of recombinant proteins at the laboratory scale for cells cultivated at low cell densities . In this study, protein splicing and in vitro cleavage of intein fusion proteins expressed in high-cell-density fed-batch fermentations of recombinant Escherichia coli were examined . Three model intein fusion constructs were used to examine the stability and splicing/cleavage activities of the fusion proteins produced under high-cell-density conditions . The data indicated that the intein fusion protein containing the wild-type intein catalyzed efficient in vivo protein splicing during high-cell-density cultivation . Also, the intein fusion proteins containing modified inteins catalyzed efficient thiol-induced in vitro cleavage reactions . The results of this study demonstrated the potential feasibility of using the intein-mediated protein purification system for industrial-scale production of recombinant proteins.

Biotechnol Prog, 2003 May-Jun, 19(3), 706 - 13
Carbon material and bioenergetic balances of xylitol production from corncobs by Debaryomyces hansenii; Rivas B et al.; The effect of oxygenation on xylitol production by the yeast Debaryomyces hansenii has been investigated in this work using the liquors from corncob hydrolysis as the fermentation medium . The concentrations of consumed substrates (glucose, xylose, arabinose, acetate and oxygen) and formed products (xylitol, arabitol, ethanol, biomass and carbon dioxide) have been used, together with those previously obtained varying the hydrolysis technique, the level of adaptation of the microorganism, the sterilization procedure and the initial substrate and biomass concentrations, in carbon material balances to evaluate the percentages of xylose consumed by the yeast for the reduction to xylitol, alcohol fermentation, respiration and cell growth . The highest xylitol concentration (71 g/L) and volumetric productivity (1.5 g/L.h) were obtained semiaerobically using detoxified hydrolyzate produced by autohydrolysis-posthydrolysis, at starting levels of xylose (S(0)) and biomass (X(0)) of about 100 g/L and 12 g(DM)/L, respectively . No less than 80% xylose was addressed to xylitol production under these conditions . The experimental data collected in this work at variable oxygen levels allowed estimating a P/O ratio of 1.16 mol(ATP)/mol(O) . The overall ATP requirements for biomass production and maintenance demonstrated to remarkably increase with X(0) and for S(0) >or= 130 g/L and to reach minimum values (1.9-2.1 mol(ATP)/C-mol(DM)) just under semiaerobic conditions favoring xylitol accumulation.

Biotechnol Prog, 2003 May-Jun, 19(3), 693 - 705
Design and installation of a next generation pilot scale fermentation system; Junker B et al.; Four new fermenters were designed and constructed for use in secondary metabolite cultivations, bioconversions, and enzyme production . A new PC/PLC-based control system also was implemented using GE Fanuc PLCs, Genius I/O blocks, and Fix Dynamics SCADA software . These systems were incorporated into an industrial research fermentation pilot plant, designed and constructed in the early 1980s . Details of the design of these new fermenters and the new control system are described and compared with the existing installation for expected effectiveness . In addition, the reasoning behind selection of some of these features has been included . Key to the design was the goal of preserving similarity between the new and previously existing and successfully utilized fermenter hardware and software installations where feasible but implementing improvements where warranted and beneficial . Examples of enhancements include strategic use of Inconel as a material of construction to reduce corrosion, piping layout design for simplified hazardous energy isolation, on-line calculation and control of nutrient feed rates, and the use of field I/O modules located near the vessel to permit low-cost addition of new instrumentation.

Appl Environ Microbiol, 2003 Jun, 69(6), 3617 - 25
Mapping of genomic regions (quantitative trait loci) controlling production and quality in industrial cultures of the edible basidiomycete Pleurotus ostreatus; Larraya LM et al.; Industrial production of the edible basidiomycete Pleurotus ostreatus (oyster mushroom) is based on a solid fermentation process in which a limited number of selected strains are used . Optimization of industrial mushroom production depends on improving the culture process and breeding new strains with higher yields and productivities . Traditionally, fungal breeding has been carried out by an empirical trial and error process . In this study, we used a different approach by mapping quantitative trait loci (QTLs) controlling culture production and quality within the framework of the genetic linkage map of P . ostreatus . Ten production traits and four quality traits were studied and mapped . The production QTLs identified explain nearly one-half of the production variation . More interestingly, a single QTL mapping to the highly polymorphic chromosome VII appears to be involved in control of all the productivity traits studied . Quality QTLs appear to be scattered across the genome and to have less effect on the variation of the corresponding traits . Moreover, some of the new hybrid strains constructed in the course of our experiments had production or quality values higher than those of the parents or other commercial strains . This approach opens the possibility of marker-assisted selection and breeding of new industrial strains of this fungus.

Appl Environ Microbiol, 2003 Jun, 69(6), 3427 - 34
1,8-dihydroxynaphthalene (DHN)-melanin biosynthesis inhibitors increase erythritol production in Torula corallina, and DHN-melanin inhibits erythrose reductase; Lee JK et al.; The yeast Torula corallina is a strong erythritol producer that is used in the industrial production of erythritol . However, melanin accumulation during culture represents a serious problem for the purification of erythritol from the fermentation broth . Melanin biosynthesis inhibitors such as 3,4-dihydroxyphenylalanine and 1,8-dihydroxynaphthalene (DHN)-melanin inhibitors were added to the T . corallina cultures . Only the DHN-melanin inhibitors showed an effect on melanin production, which suggests that the melanin formed during the culturing of T . corallina is derived from DHN . This finding was confirmed by the detection of a shunt product of the pentaketide pathway, flaviolin, and elemental analysis . Among the DHN-melanin inhibitors, tricyclazole was the most effective . Supplementation with tricyclazole enhanced the production of erythritol while significantly inhibiting the production of DHN-melanin and DHN-melanin biosynthetic enzymes, such as trihydroxynaphthalene reductase . The erythrose reductase from T . corallina was purified to homogeneity by ion-exchange and affinity chromatography . Purified erythrose reductase was significantly inhibited in vitro in a noncompetitive manner by elevated levels of DHN-melanin . In contrast, the level of erythrose reductase activity was unaffected by increasing concentrations of tricyclazole . These results suggest that supplemental tricyclazole reduces the production of DHN-melanin, which may lead to a reduction in the inhibition of erythrose reductase and a higher yield of erythritol . This is the first report to demonstrate that melanin biosynthesis inhibitors increase the production of a sugar alcohol in T . corallina.

Biosens Bioelectron, 2003 Aug 15, 18(9), 1125 - 34
Improved selectivity of microbial biosensor using membrane coating . Application to the analysis of ethanol during fermentation; Tkac J et al.; A ferricyanide mediated microbial biosensor for ethanol detection was prepared by surface modification of a glassy carbon electrode . The selectivity of the whole Gluconobacter oxydans cell biosensor for ethanol determination was greatly enhanced by the size exclusion effect of a cellulose acetate (CA) membrane . The use of a CA membrane increased the ethanol to glucose sensitivity ratio by a factor of 58.2 and even the ethanol to glycerol sensitivity ratio by a factor of 7.5 compared with the use of a dialysis membrane . The biosensor provides rapid and sensitive detection of ethanol with a limit of detection of 0.85 microM (S/N=3) . The selectivity of the biosensor toward alcohols was better compared to previously published enzyme biosensors based on alcohol oxidase or alcohol dehydrogenases . The biosensor was successfully used in an off-line monitoring of ethanol during batch fermentation by immobilized Saccharomyces cerevisiae cells with an initial glucose concentration of 200 g l(-1).

Environ Toxicol Chem, 2003 Jun, 22(6), 1214 - 20
Reductive dechlorination of polychlorinated biphenyls in sediment from the Twelve Mile Creek arm of Lake Hartwell, South Carolina, USA; Pakdeesusuk U et al.; Lake Hartwell is a U.S . Army Corps of Engineers reservoir system located on the state line between South Carolina and Georgia, USA . The lake was contaminated with an estimated 200 metric tons of polychlorinated biphenyls ({PCBs}; mainly Aroclor 1016 and 1254), and the entire Twelve Mile Creek watershed and the Seneca River arm of Lake Hartwell were placed on the National Priorities List . Monitored natural attenuation was chosen as a remedy for the contaminated sediment . The relatively warm temperature of Lake Hartwell and lack of significant cocontaminants along with the PCBs distinguish this site from others that have been studied for microbially mediated reductive dechlorination . Microcosm studies were conducted with sediment from two locations in the Twelve Mile Creek arm and confirmed the presence of indigenous microorganisms capable of reductively dechlorinating Aroclor 1254, which contains predominantly tetra-, penta-, and hexachlorobiphenyl . The average number of total chlorines per biphenyl decreased from 4.8 to 4.9 to 2.9 to 3.0, following 250 to 260 d of incubation . The maximum observed dechlorination rates were 0.29 to 0.87 microg-atoms Cl- per gram sediment dry weight per week . The onset of dechlorination activity correlated strongly with maximum methanogenesis, which occurred without a lag in samples from the site that showed signs of in situ fermentation activity . Dechlorination occurred primarily at the meta and para positions (58-63% removal), with no apparent decrease in ortho chlorines . This most closely resembles pattern M, characterized by preferential removal of unflanked and flanked meta chlorines . The microcosm results are consistent with sediment cores analyzed from the same locations, which indicate accumulation with depth of the same ortho- and para-substituted congeners . It therefore appears that the success of monitored natural attenuation for Lake Hartwell will hinge on covering the recalcitrant PCBs with a sufficient amount of uncontaminated sediment to isolate them from the food chain.

Antonie Van Leeuwenhoek, 2003, 83(2), 149 - 54
Yeasts present during spontaneous fermentation of Lake Erie Chardonnay, Pinot Gris and Riesling; van Keulen H et al.; The composition of wine yeast populations, present during spontaneous fermentation of Chardonnay, Pinot Gris and Riesling from the Lake Erie Region was studied . A combination of biochemical and molecular techniques was used to identify non-Saccharomyces and Saccharomyces yeast isolates . The biochemical techniques included analysis of yeast isolates by sugar fermentation and carbon and nitrogen assimilation . Molecular techniques involved ribotyping of a highly variable segment in the 26S rRNA gene using DNA sequence analysis and restriction fragment length polymorphism of amplified DNA . The results show that of the non-Saccharomyces yeasts, several related species of Hanseniaspora, were the most abundant yeasts present during early stages of fermentation . Later in fermentation S . cerevisiae dominated, and based on biochemical analyses consisted of a heterogeneous group of genotypes . There were no major differences in yeast populations among the three types of juice analyzed.

Int Rev Neurobiol, 2003, 54, 199 - 228
Drosophila melanogaster, a genetic model system for alcohol research; Guarnieri DJ et al.; In its natural environment, which consists of fermenting plant materials, the fruit fly Drosophila melanogaster encounters high levels of ethanol . Flies are well equipped to deal with the toxic effects of ethanol; they use it as an energy source and for lipid biosynthesis . The primary ethanol-metabolizing pathway in flies involves the enzymes alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH); their role in adaptation to ethanol-rich environments has been studied extensively . The similarity between Drosophila and mammals is not restricted to the manner in which they metabolize ethanol; behaviors elicited by ethanol exposure are also remarkably similar in these organisms . Flies show signs of acute intoxication, which range from locomotor stimulation at low doses to complete sedation at higher doses, they develop tolerance upon intermittent ethanol exposure, and they appear to like ethanol, showing preference for ethanol-containing media . Molecular genetic analysis of ethanol-induced behaviors in Drosophila, while still in its early stages, has already revealed some surprising parallels with mammals . The availability of powerful tools for genetic manipulation in Drosophila, together with the high degree of conservation at the genomic level, make Drosophila a promising model organism to study the mechanism by which ethanol regulates behavior and the mechanisms underlying the organism's adaptation to long-term ethanol exposure.

Biosci Biotechnol Biochem, 2003 Apr, 67(4), 765 - 71
Analysis of the pyruvate permease gene (JEN1) in glucose derepression yeast (Saccharomyces cerevisiae) Isolated from a 2-deoxyglucose-tolerant mutant, and its application to sake making; Tsuboi H et al.; We isolated mutants of S . cerevisiae in which expression of the JEN1 gene encoding a pyruvate transporter was insensitive to glucose repression . The isolated mutant GDR19 expressed JEN1 and absorbed pyruvate in the presence of glucose . In a DNA microarray analysis, GDR19 highly expressed many more genes, including JEN1, in the presence of glucose compared with the parental strain B29 . Some of these genes are under the control of the transcription factor Mig1p and are normally repressed in the presence of glucose . The concentrations of organic acids in sake made with GDR19 were different from those in sake made with B29 . Changes in the pyruvate concentration in the sake mash made with GDR19 were not very different from those in sake mash made with B29, and both GDR19 and B29 expressed JEN1 during fermentation . When the ethanol concentration was over 2%, JEN1 expression in B29 was similar in the presence and absence of glucose . The expression of JEN1 in sake mash in spite of the presence of glucose appeared to be caused by the coexistence of ethanol.

Biotechnol Bioeng, 2003 Aug 5, 83(3), 312 - 20
Inhibitory effect of carbon dioxide on the fed-batch culture of Ralstonia eutropha: evaluation by CO2 pulse injection and autogenous CO2 methods; Shang L et al.; In order to see the effect of CO(2) inhibition resulting from the use of pure oxygen, we carried out a comparative fed-batch culture study of polyhydroxybutyric acid (PHB) production by Ralstonia eutropha using air and pure oxygen in 5-L, 30-L, and 300-L fermentors . The final PHB concentrations obtained with pure O(2) were 138.7 g/L in the 5-L fermentor and 131.3 g/L in the 30-L fermentor, which increased 2.9 and 6.2 times, respectively, as compared to those obtained with air . In the 300-L fermentor, the fed-batch culture with air yielded only 8.4 g/L PHB . However, the maximal CO(2) concentrations in the 5-L fermentor increased significantly from 4.1% (air) to 15.0% (pure O(2)), while it was only 1.6% in the 30-L fermentor with air, but reached 14.2% in the case of pure O(2) . We used two different experimental methods for evaluating CO(2) inhibition: CO(2) pulse injection and autogenous CO(2) methods . A 10 or 22% (v/v) CO(2) pulse with a duration of 3 or 6 h was introduced in a pure-oxygen culture of R . eutropha to investigate how CO(2) affects the synthesis of biomass and PHB . CO(2) inhibited the cell growth and PHB synthesis significantly . The inhibitory effect became stronger with the increase of the CO(2) concentration and pulse duration . The new proposed autogenous CO(2) method makes it possible to place microbial cells under different CO(2) level environments by varying the gas flow rate . Introduction of O(2) gas at a low flow rate of 0.42 vvm resulted in an increase of CO(2) concentration to 30.2% in the exit gas . The final PHB of 97.2 g/L was obtained, which corresponded to 70% of the PHB production at 1.0 vvm O(2) flow rate . This new method measures the inhibitory effect of CO(2) produced autogenously by cells through the entire fermentation process and can avoid the overestimation of CO(2) inhibition without introducing artificial CO(2) into the fermentor .

Biotechnol Bioeng, 2003 Aug 5, 83(3), 293 - 302
Model-based analysis of anaerobic acetate uptake by a mixed culture of polyphosphate-accumulating and glycogen-accumulating organisms; Zeng RJ et al.; An increasing number of studies shows that the glycogen-accumulating organisms (GAOs) can survive and may indeed proliferate under the alternating anaerobic/aerobic conditions found in EBPR systems, thus forming a strong competitor of the polyphosphate-accumulating organisms (PAOs) . Understanding their behaviors in a mixed PAO and GAO culture under various operational conditions is essential for developing operating strategies that disadvantage the growth of this group of unwanted organisms . A model-based data analysis method is developed in this paper for the study of the anaerobic PAO and GAO activities in a mixed PAO and GAO culture . The method primarily makes use of the hydrogen ion production rate and the carbon dioxide transfer rate resulting from the acetate uptake processes by PAOs and GAOs, measured with a recently developed titration and off-gas analysis (TOGA) sensor . The method is demonstrated using the data from a laboratory-scale sequencing batch reactor (SBR) operated under alternating anaerobic and aerobic conditions . The data analysis using the proposed method strongly indicates a coexistence of PAOs and GAOs in the system, which was independently confirmed by fluorescent in situ hybridization (FISH) measurement . The model-based analysis also allowed the identification of the respective acetate uptake rates by PAOs and GAOs, along with a number of kinetic and stoichiometric parameters involved in the PAO and GAO models . The excellent fit between the model predictions and the experimental data not involved in parameter identification shows that the parameter values found are reliable and accurate . It also demonstrates that the current anaerobic PAO and GAO models are able to accurately characterize the PAO/GAO mixed culture obtained in this study . This is of major importance as no pure culture of either PAOs or GAOs has been reported to date, and hence the current PAO and GAO models were developed for the interpretation of experimental results of mixed cultures . The proposed method is readily applicable for detailed investigations of the competition between PAOs and GAOs in enriched cultures . However, the fermentation of organic substrates carried out by ordinary heterotrophs needs to be accounted for when the method is applied to the study of PAO and GAO competition in full-scale sludges .

Int J Food Microbiol, 2003 Jul 15, 84(1), 33 - 9
Molecular typing techniques as a tool to differentiate non-Saccharomyces wine species; Capece A et al.; A total of 32 yeast strains belonging to four non-Saccharomyces species associated with winemaking was characterized by different molecular techniques . The PCR amplification of 18S rRNA-coding DNA and nontranscribed spacer, followed by restriction analysis with the endonucleases HaeIII and MspI, and PCR fingerprinting with microsatellite primers (GAC)(5) and (GTG)(5) were used . The methods used provided species-specific profiles and proved to be fast and reliable for monitoring the evolution of the four non-Saccharomyces yeast populations throughout wine fermentation.

Int J Food Microbiol, 2003 Jul 15, 84(1), 21 - 6
Yeast autolytic mutants potentially useful for sparkling wine production; Gonzalez R et al.; A selection method, based on a temperature-sensitive autolytic phenotype, has been used to genetically improve a second fermentation Saccharomyces cerevisiae yeast strain by UV mutagenesis . The mutations carried by the resulting strains affected cell morphology, growth kinetics, sporulation and the release of nitrogenous compounds in an accelerated autolysis experimental model . Their fermentation power was not severely impaired.

Nutrition, 2003 Jun, 19(6), 524 - 30
Making bread with sourdough improves mineral bioavailability from reconstituted whole wheat flour in rats; Lopez HW et al.; OBJECTIVE: We compared the effects of different kinds of bread fermentation on mineral bioavailability . METHODS: Wistar rats were fed one of the following experimental diets for 21 d: control, reconstituted whole wheat flour (white flour plus bran), yeast bread, and sourdough bread . The apparent mineral absorption and intestinal fermentation were measured in each animal . RESULTS: Phytate contents in yeast and sourdough bread were lower than in reconstituted whole wheat flour (-52% and -71%, respectively) . Total cecal pool of short-chain fatty acids, in particular the butyrate pool, was significantly increased by the ingestion of unrefined products . Calcium homeostasis was not modified by these nutritional conditions, whereas magnesium absorption was significantly greater in rats fed the control and sourdough diets than in those consuming whole wheat flour and yeast bread . Magnesium kidney excretion was slightly stimulated by sourdough bread . Compared with the control diet, iron balance was significantly reduced by reconstituted whole wheat flour diet . Yeast bread making counteracted the deleterious effects of whole wheat on iron absorption, whereas sourdough bread making enhanced iron absorption . Further, liver and plasma iron and transferrin saturation levels were lower in rats adapted to the flour diet than in other groups . Zinc absorption was strongly depressed in the presence of unprocessed reconstituted whole wheat flour in the diet, but yeast fermentation afforded a zinc assimilation comparable to the control diet, whereas the sourdough bread led to maximal zinc absorption . Copper absorption increased significantly when rats were fed the sourdough bread, whereas unprocessed whole flour depressed copper absorption (-41% versus control diet) . CONCLUSION: Mineral bioavailability from reconstituted whole wheat flour can be improved by bread making . Although yeast fermentation minimizes the unfavorable effects of phytic acid, sourdough bread is a better source of available minerals, especially magnesium, iron, and zinc.

Mol Biotechnol, 2003 Jul, 24(3), 257 - 81
A new look at xylanases: an overview of purification strategies; Sa-Pereira P et al.; Interest in xylanases from different sources has increased markedly in the past decade, in part because of the application of these enzymes in the pulp and paper industry . Purity and purification costs are becoming important issues in modern biotechnology as the industry matures and competitive products reach the marketplace . Thus, new paths for successful and efficient xylanase recovery have to be followed . This article reviews the isolation and purification methods used for the recovery of microbial xylanases . Origins and applications of xylanases are described, highlighting the special features of this class of enzymes, such as the carbohydrate-binding domains (CBDs) and their importance in the development of affinity methodologies to increase and facilitate xylanase purification . Implications of recombinant DNA technology for the isolation and purification of xylanases are evaluated . Several purification procedures are analyzed, taking into consideration the sequence of the methods used in each and the number of times each method is used . New directions to improve xylanase separation and purification from fermentation media are described.

Mol Biol Evol, 2003 Aug, 20(8), 1310 - 7 Epub 2003 May 30.
Parallel functional changes in the digestive RNases of ruminants and colobines by divergent amino acid substitutions; Zhang J; A morphological or physiological trait may appear multiple times in evolution . At the molecular level, similar protein functions may emerge independently in different lineages . Whether these parallel functional changes are due to parallel amino acid substitutions has been a subject of debate . Here, I address this question using digestive ribonucleases (RNases) of two groups of foregut-fermenting mammals: ruminant artiodactyls and colobine monkeys . The RNase1 gene was duplicated twice in ancestral ruminants at least 40 MYA, and it was also duplicated in the douc langur, an Asian colobine, approximately 4 MYA . After duplication, similar functional changes occurred in the ruminant and monkey enzymes . Interestingly, five amino acid substitutions in ruminant RNases that are known to affect its catalytic activity against double-stranded (ds) RNA did not occur in the monkey enzyme . Rather, a similar functional change in the monkey was caused by a different set of nine substitutions . Site-directed mutagenesis was used to make three of the five ruminant-specific substitutions in the monkey enzyme . Functional assays of these mutants showed that one of the three substitutions has a similar effect in monkeys, the second has a stronger effect, and the third has an opposite effect . These results suggest that (1) an evolutionary problem can have multiple solutions, (2) the same amino acid substitution may have opposite functional effects in homologous proteins, (3) the stochastic processes of mutation and drift play an important role even at functionally important sites, and (4) protein sequences may diverge even when their functions converge.

Microbiology, 2003 Jun, 149(Pt 6), 1523 - 32
Changes in GE2270 antibiotic production in Planobispora rosea through modulation of methylation metabolism; Gastaldo L et al.; Thiazolylpeptide GE2270 is a potent antibiotic inhibiting protein synthesis in Gram-positive bacteria . It is produced as a complex of 10 related metabolites, differing mainly in the degree of methylation, by fermentation of the rare actinomycete Planobispora rosea ATCC 53773 . Addition of vitamin B12 to the fermentation medium doubled total complex production and markedly changed the relative production of the various GE2270 metabolites, enhancing the biosynthesis of the more methylated component A . Among methylation inhibitors, the addition of sinefungin increased the amount of factor D2, which differs from component A in the lack of a methyl group . Since sinefungin is an S-adenosyl-L-methionine methyltransferase-specific inhibitor, these results indicate that the methylation step converting D2 into A involves an S-adenosyl-L-methionine methyltransferase . Simultaneous supplementation of vitamin B12 and sinefungin led to a twofold increase in D2 concentration, showing that vitamin B12, in addition to having an effect on the late methylation step, exerts a stimulating action on antibiotic backbone synthesis . This is possibly due to its role in an unusual pathway of serine synthesis peculiar to P . rosea metabolism . Finally, fermentation medium modifications were shown to be useful for the production of industrially valuable levels of components A or D2 in the GE2270 complex as starting points for the production of new interesting semi-synthetic antibiotics.

Antonie Van Leeuwenhoek, 2003, 83(4), 333 - 40
Characterization of a non-pigment producing Monascus purpureus mutant strain; Rasheva TV et al.; A characterization of a non-pigment producing mutant Monascus purpureus M12 compared with its parental strain Monascus purpureus Went CBS 109.07 has been performed aiming to investigate the relation between pigment biosynthesis and other characteristics of these fungi . A comparison has been made of morphological features, some physiological properties and biochemical activities of both strains . The albino mutant exhibits an anamorph life cycle, high conidia forming capability, slower radial growth rate and temperature sensitivity . The assimilation capacity of both strains for mono-, disaccharides and some alcohols is in the same range (Yx/c 0.2 - 0.35), while the red strain has a higher fermentation capacity . In a selected albino mutant, the growth rate, metabolic activity and capacity for production of typical for Monascus fungi secondary metabolites were reduced considerably . Hydrolytic activity towards natural substrates expressed through glucoamylase and protease was approximately 10 fold lower in the non pigment producing strain (0.05 - 0.08 U/mg protein and 0.01 - 0.07 U/mg protein respectively) compared with the red one . Important qualitative differences between both strains was found in fatty acid composition and in the production of citrinin and monacolin . The mutant strain possessed C17, C20 and C22 fatty acids and did not produce citrinin.

Immunobiology, 2003, 207(3), 223 - 33
Synthetic mycoplasma-derived lipopeptide MALP-2 induces maturation and function of dendritic cells; Weigt H et al.; Dendritic cells (DC) modulate immune responses depending on the nature of the antigens . Receptors capable of discriminating these antigens on the basis of the pathogen-associated molecular patterns (PAMP) belong to the Toll-like receptor (TLR) family . The macrophage-activating lipopeptide 2 kDa (MALP-2), a synthetic lipopeptide derived from Mycoplasma fermentans, signals through TLR-2 and TLR-6 . The aim of this study was to examine whether MALP-2 can modulate the functional properties of human monocyte-derived DC . The effects of this treatment were compared to those of the TLR-4 agonist lipopolysaccharide (LPS) . To ensure clinical applicability, DC were generated under serum-free conditions . MALP-2 and LPS stimulation induced the expression of CD83 and increased the expressions of CD80, CD86, HLA-ABC and CD40 . Furthermore, both substances decreased the endocytotic capacity of DC and induced the release of bioactive TNF-alpha and IL-10, whereas LPS additionally increased IL-12 release . Pretreatment with both substances boosted the allostimulatory capacity of DC . In a coculture with autologous lymphocytes, either MALP-2 or LPS pretreated DC induced a marked proliferation of lymphocytes, but only DC prestimulated with MALP-2 activated lymphocytes to produce the cytokines IL-4, IL-5 and IFN-gamma . No polarisation of lymphocytes into T-helper (Th)1 or Th2 was detected . These data indicate that MALP-2 is a potential candidate to modulate DC for clinical applications.

Zhongguo Zhong Yao Za Zhi, 2001 Sep, 26(9), 622 - 5
{Effects of fermented Cordyceps powder on pulmonary function in sensitized guinea pigs and airway inflammation in sensitized rats}; Lin XX et al.; OBJECTIVE: To study Cordyceps (artificial fermented Cordyceps sinensis(Berk.) Sacc) powderin the treatment of asthma in the animal models . METHOD: Pulmonary function and airway inflammation in vivo were investigated . RESULT: Cordyceps, 5g.kg-1(ig), significantly inhibited bronchial challenge of ovalbumin-induced change of RL and Cdyn (P < 0.05) and inhibited antigen-induced increase of eosinophils in the BALF of rats (P < 0.05) . CONCLUSION: The results suggested cordyceps could be applied for the prevention and cure of asthma.

Zhongguo Zhong Yao Za Zhi, 2001 Dec, 26(12), 831 - 4
{A comparative study on sterols of ethanol extract and water extract from Hericium erinaceus}; Li JL et al.; OBJECTIVE: To study the pharmaceutic chemical basis of the different medicinal effects, and to compare and analyze the sterols in Hericium erinaceus mycelia and its ethanol extract, water extract derived from solid fermented mycelia . METHOD: The components of Hericium erinaceus mycelia and two kinds of extract were compared with some biochemical methods such as GC, RP-HPLC, etc . Sterol composition in Hericium erinaceus mycelia was determined by using GC-mass spectrometry . RESULTS: The content of crude polysaccharide, water-soluble protein and fatty acid of ethanol extract showed no obvious advantage over its water extract . However, there was significant difference between two kinds of extract in the content of sterols . And ergostane derivatives, beta-sitosterol and C28 sterol with four bonds were detected as the major sterols in Hericium erinaceus mycelia, among which ergosterol was the principal sterol . CONCLUSION: Sterols in Hericium erinaceus exist mainly in ethanol extract . And three sterols are found for the first time from Hericium erinaceus, which are ergostane derivatives, beta-sitosterol and C28 sterol with four bonds.

J Bacteriol, 2003 Jun, 185(12), 3596 - 605
Escherichia coli O157:H7 Shiga toxin-encoding bacteriophages: integrations, excisions, truncations, and evolutionary implications; Shaikh N et al.; As it descended from Escherichia coli O55:H7, Shiga toxin (Stx)-producing E . coli (STEC) O157:H7 is believed to have acquired, in sequence, a bacteriophage encoding Stx2 and another encoding Stx1 . Between these events, sorbitol-fermenting E . coli O157:H(-) presumably diverged from this clade . We employed PCR and sequence analyses to investigate sites of bacteriophage integration into the chromosome, using evolutionarily informative STEC to trace the sequence of acquisition of elements encoding Stx . Contrary to expectations from the two currently sequenced strains, truncated bacteriophages occupy yehV in almost all E . coli O157:H7 strains that lack stx(1) (stx(1)-negative strains) . Two truncated variants were determined to contain either GTT or TGACTGTT sequence, in lieu of 20,214 or 18,895 bp, respectively, of the bacteriophage central region . A single-nucleotide polymorphism in the latter variant suggests that recombination in that element extended beyond the inserted octamer . An stx(2) bacteriophage usually occupies wrbA in stx(1)(+)/stx(2)(+) E . coli O157:H7, but wrbA is unexpectedly unoccupied in most stx(1)-negative/stx(2)(+) E . coli O157:H7 strains, the presumed progenitors of stx(1)(+)/stx(2)(+) E . coli O157:H7 . Trimethoprim-sulfamethoxazole promotes the excision of all, and ciprofloxacin and fosfomycin significantly promote the excision of a subset of complete and truncated stx bacteriophages from the E . coli O157:H7 strains tested; bile salts usually attenuate excision . These data demonstrate the unexpected diversity of the chromosomal architecture of E . coli O157:H7 (with novel truncated bacteriophages and multiple stx(2) bacteriophage insertion sites), suggest that stx(1) acquisition might be a multistep process, and compel the consideration of multiple exogenous factors, including antibiotics and bile, when chromosome stability is examined.

Zhongguo Zhong Yao Za Zhi, 2002 Jun, 27(6), 431 - 4
{A new steroidal glycoside from fermented leaves of Agave americana}; Jin JM et al.; OBJECTIVE: To investigate the constituents of fermented leaves of Agave americana, and discover new compounds . METHOD: Compounds were purified with silica gel and C8 reverse--phase silica gel column chromatography . The structures were elucidated by chemical and spectroscopic evidence . RESULT: Three steroidal compounds were obtained and their structures were identified as (25R)-5 alpha-spirostan-3 beta, 6 alpha, 23 alpha-triol 6-O-beta-D-glucopyranoside(1), (25R)-5 alpha-spirostan-3 beta, 6 alpha, 23 alpha-triol-3, 6-di-O-beta-D-glucopyranoside (cantalasaponin-1) (2) and (25R)-5 alpha-spirostan-3 beta, 6 alpha, 23 alpha-triol(hongguanggenin) (3) . CONCLUSION: Compound 1 is new compound, named agamenoside C.

J Anim Sci, 2003 May, 81(5), 1210 - 9
Effects of two different dietary fermentable carbohydrates on activity and heat production in group-housed growing pigs; Rijnen MM et al.; The effects of two sources of dietary fiber (DF) on behavior and heat production (HP) in group-housed growing pigs were studied . Twenty clusters of 14 barrows (50 kg) were fed one of 10 diets . Diets differed mainly in type and content of fermentable DF (fDF) and in content of digestible starch . Five diets contained solvent-extracted coconut meal (SECM) and five diets contained soybean hulls (SBH) as the main fDF source . On an as-fed basis, pigs received 3.5, 13.2, 23.0, 32.7, or 42.4 g x kg(-0.75) x d(-1) of SECM or SBH . A total of 280 crossbred growing pigs were used, divided into clusters of 14 pigs each . Pigs were group-housed and fed at 2.5 times the assumed maintenance energy requirements . All clusters were fed similar amounts of NE, ileal-digestible protein and amino acids, vitamins, and minerals . Consequently, DMI differed among diets because NE content decreased with increasing DF content . After a 32-d preliminary period, HP was measured per cluster during a 7-d experimental period in environmentally controlled respiration chambers . Behavior of the pigs was recorded using time-lapse video recordings during two different days within the experimental period . Intake of digestible starch and fDF was different (P < 0.001) among diets, whereas intake of digestible CP was similar among diets . On average, pigs spent 153 min standing, 42 min sitting, 202 min lying on their chest, and 1,043 min lying on their flanks each day . Pigs fed SECM diets spent, on average, less time (P < 0.05) lying on their chest than pigs fed SBH diets . Total time spent on physical activity (i.e., standing plus sitting, 195 min/d) was not affected by diet . Total HP and resting HP were affected by diet and were on average lower (P < 0.01) for pigs fed SECM diets than for pigs fed SBH diets . Activity-related heat production (AHP) averaged 65 kJ x kg(-0.75) x d(-1) and was not affected by diet . There was a linear relationship (P < 0.001) between fDF intake and HP, but there was no relationship between fDF intake and AHP . During different parts of the day, fDF intake also affected HP . The saving effect of physical activity on the NE values of fDF from SECM and SBH were 0.56 and 0.84 kJ/g of fDF intake, respectively . Neither of these saving effects was significantly different from zero . We conclude that fDF from SECM and SBH did not affect energy expended on physical activity by growing pigs, and that the NE value of fDF from SECM and SBH was not affected by changes in physical activity.

J Biotechnol, 2003 Jun 12, 103(1), 87 - 91
Monitoring batch fermentations with an electronic tongue; Turner C et al.; An electronic tongue comprising 21 potentiometric chemical sensors with pattern recognition tools was used for the rapid off-line monitoring of batch Escherichia coli fermentations . The electronic tongue was capable of monitoring the changes in the media composition as the fermentation progressed, and could correlate this with an increase in biomass . The electronic tongue was also able to monitor the increase in organic acids, especially acetic acid, throughout the fermentation . This technique clearly shows promise as a rapid tool for fermentation monitoring.

J Insect Physiol, 1998 May, 44(5-6), 427 - 436
Environmental hypoxia affects osmotic and ionic regulation in freshwater midge-larvae; Scholz F et al.; The effect of anaerobic metabolism on the osmotic and ionic regulation of the extracellular fluid was examined . Larvae of three species, characterized by different hypoxia tolerance, were studied: Chaoborus crystallinus, Culex pipiens and Chironomus gr . plumosus . The use of the capillary electrophoresis technique made it possible to determine approximately 15 different ions from individual hemolymph samples . The hemolymph concentration of both inorganic and organic anions and cations as well as the osmolality were measured . A correlation between the hypoxia tolerance and the capability to avoid net changes in the ion concentration or in the osmolality of the three species studied here is proposed: Culex larvae, which have the lowest hypoxia tolerance, show a very large and very rapid lactate accumulation in their hemolymph under experimental hypoxia . This lactate accumulation is not compensated for by a change in the concentration of any other ion . Chaoborus larvae, with a medium hypoxia tolerance, utilize their very large hemolymph malate pool as a source of anaerobic energy . It is converted into succinate, thus inducing little net changes in the sum of the anions . There is a marked increase of the hemolymph osmolality, though . Chironomus larvae have the highest hypoxia tolerance and there are remarkably little changes in their hemolymph under hypoxia . Although these larvae are described as relying mainly on ethanol fermentation under environmental anaerobiosis, we demonstrated a marked lactate fermentation in severe hypoxia . The lactate accumulation observed in our study was compensated by a concomittant decrease of the hemolymph chloride concentration.

J Insect Physiol, 2002 Oct, 48(10), 945 - 950
Semiochemicals from ovaries of gravid females attract ovipositing female houseflies, Musca domestica; Jiang Y et al.; Chemical signals originating from the ovaries of gravid females of Musca domestica (Diptera: Cyclorrhapha: Muscidae) attract ovipositing females to common egg-laying sites . Behavioral experiments indicated that females preferred to oviposit in fermented wheat bran containing ovaries from reproductively mature houseflies . Females preferred to oviposit in fermented wheat bran than wet wheat bran . This effect was additive with the attraction to housefly ovaries . Solvent extracts from housefly ovaries were attractive to gravid females . Extracts obtained with hexane were most attractive to gravid females for egg laying, and extracts obtained with ethyl acetate attracted more egg laying than extracts obtained by dichloromethane . Gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC-MS) analysis showed that tricosane and (Z)-9-tricosene were the main components of the hexane extracts . Both tricosane and (Z)-9-tricosene were shown to elicit dose-dependent aggregation of gravid females in oviposition bioassays, but high doses of either chemical were not attractive.

Curr Pharm Des, 2003, 9(16), 1297 - 308
Food-derived bioactive peptides--opportunities for designing future foods; Korhonen H et al.; Dietary proteins are known to carry a wide range of nutritional, functional and biological properties . Nutritionally, the proteins are a source of energy and amino acids, which are essential for growth and maintenance . Functionally, the proteins contribute to the physicochemical and sensory properties of various protein-rich foods . Furthermore, many dietary proteins possess specific biological properties which make these components potential ingredients of functional or health-promoting foods . Many of these properties are attributed to physiologically active peptides encrypted in protein molecules . Particularly rich sources of such peptides are milk and egg, but they are also found in meat of various kinds as well as many plants . These peptides are inactive within the sequence of parent protein and can be released during gastrointestinal digestion or food processing . Depending on the amino acid sequence, these peptides may exert a number of different activities in vivo, affecting, e.g., the cardiovascular, endocrine, immune and nervous systems in addition to nutrient utilization . There is increasing commercial interest in the production of bioactive peptides from various sources . Industrial-scale production of such peptides is, however, hampered by the lack of suitable technologies . Bioactive peptides can also be produced from milk proteins through fermentation of milk, by starters employed in the manufacture of fermented milks or cheese . In particular, antihypertensive peptides have been identified in fermented milk, whey and ripened cheese . A few of these peptides have been commercialised in the form of fermented milks . There is a need to develop technologies which retain or even enhance the activity of bioactive peptides in food systems . Also, it is essential to study the optimum utilization of such peptides during passage through the gastrointestinal tract.

J Agric Food Chem, 2003 Jun 4, 51(12), 3682 - 7
Pathway leading to the formation of anthocyanin-vinylphenol adducts and related pigments in red wines; Schwarz M et al.; On the basis of observations from Vitis vinifera cv . Pinotage wines and experiments performed in model wine medium, a new chemical pathway responsible for the formation of anthocyanin-vinylphenol adducts in red wines is described . Until now, these pigments have been considered to be reaction products of anthocyanins and vinylphenols, the latter being generated during fermentation by enzymatic decarboxylation of the respective cinnamic acids . The mechanism of the novel pathway, involving intact hydroxycinnamic acid and anthocyanin, is explained . Only cinnamic acids with electron-donating substituents on the aromatic ring, such as coumaric acid, ferulic acid, caffeic acid, and sinapic acid, undergo this conversion, as they stabilize an intermediately formed carbenium ion . Decarboxylation and oxidation of the pyran moieties are the final steps in the generation of the corresponding 4-vinylphenol, 4-vinylguaiacol, 4-vinylcatechol, and 4-vinylsyringol adducts of anthocyanins in red wine.

Anal Sci, 2003 May, 19(5), 659 - 64
Spectrophotometric assay of yeast vitality using 2,3,5,6-tetramethyl-1,4-benzoquinone and tetrazolium salts; Tsukatani T et al.; A method for the spectrophotometric assay of yeast vitality was developed using 2,3,5,6-tetramethyl-1,4-benzoquinone and tetrazolium salts . The metabolic efficiency of 2,3,5,6-tetramethyl-1,4-benzoquinone by yeast cells was used as an index of yeast vitality . 2,3,5,6-Tetramethyl-1,4-benzoquinone was reduced to 2,3,5,6-tetramethyl-1,4-hydroquinone by yeast cells . Then, the superoxide anion radicals generated from O2 by reduction with 2,3,5,6-tetramethyl-1,4-hydroquinone under alkaline conditions reduced tetrazolium salts to formazan, which exhibited absorbance maxima at 440 nm . A linear relationship between the absorbance and viable cell density was obtained in the range of 1.0 x 10(5)-2.0 x 10(7) cells/ml for a sample solution . During the cultivation of yeast cells, the absorbance showed almost an anti-parallel change with that of glucose in yeast growth and fermentation, suggesting that the absorbance change reflected the vitality of yeast cells.

Biotechnol Bioeng, 2003 Jul 20, 83(2), 235 - 40
Vibrating membrane filtration for recovery and concentration of insect killing nematodes; Wilson JA et al.; New experimental data are reported that demonstrate the use of a novel vibrating membrane filter (VMF) for the combined recovery and concentration of two species of nematodes, S . feltiae and P . hermaphrodita, from mature liquid fermentation cultures . The disk membrane module had a working surface area of 0.2 m(2) and was operated at a constant flow rate of 0.2 m(3) h(-1) . The recovery of the viable nematodes from the spent media and nonviable nematodes was assisted by an independently imposed oscillatory motion of the disk assembly, which produced an intense shear field at the membrane surface with calculated mean values on the order of 10(4) s(-1) . Adult (nonviable) nematodes in the fermentation culture were preferentially dissolved in a detergent (sodium dodecylsulfate) and successfully separated from the juveniles using the VMF equipment . Permeate fluxes on the order of 15 to 30 L/m(2/)h were achieved for an operating transmembrane pressure of 800 mbar . Industrial-scale liquid fermentation for the manufacture of nematodes as biopesticides produces the viable nematode life stages in low-concentration suspension containing large quantities of spent media and other waste material . The VMF equipment provided a flexible operation for separation, cleaning, and concentration of viable nematodes from the fermentation broths .

Biotechnol Bioeng, 2003 Jul 20, 83(2), 191 - 200
Fermentation of corn stover to carboxylic acids; Thanakoses P et al.; This article describes countercurrent fermentation to anaerobically convert corn stover and pig manure to mixed carboxylic acids using a mixed culture of mesophilic microorganisms . Corn stover was pretreated with lime to increase digestibility . The Continuum Particle Distribution Model (CPDM) was used to simulate continuous fermentors based on data collected from batch experiments . This model saves considerable time in determining optimum operating conditions . For 80% corn stover/20% pig manure, the highest total carboxylic acid productivity was 1.81 g/(L of liquid . d) at a concentration of 21.4 g total acid/L . The highest total acid selectivity, yield, and conversion were 0.714 g total acid/g volatile solids (VS) digested, 0.550 g total acid/g VS fed, and 0.770 g VS digested/g VS fed, respectively, at a concentration of 16.0 g total acid/L . CPDM predicted the acid concentration and conversion within 13.4 and 11.6%, respectively .

Biotechnol Bioeng, 2003 Jul 20, 83(2), 181 - 90
Constitutive versus thermoinducible expression of heterologous proteins in Escherichia coli based on strong PR,PL promoters from phage lambda; Menart V et al.; Constitutive and thermoinducible expression plasmids based on strong P(R),P(L) promoters from phage lambda were compared for production of TNF-alpha and its analogs under various conditions . Much higher accumulation of TNF was obtained in a constitutive system, so the wider applicability of such systems was studied . In constitutive systems, proteolytically susceptible proteins can be produced easily at low cultivation temperatures and the addition of expensive or toxic chemical inducers is not required . On the other hand, toxic proteins cannot be produced and selection pressure must be strictly maintained to ensure segregational stability of plasmids . Accumulation of TNF-alpha and various analogs at levels up to 25% of total soluble protein was repeatedly achieved, which was 2-3-fold higher than in a thermoinducible system . The stable behavior of the constitutive system in laboratory fermentors was also confirmed . We propose the constitutive system described here as a general model for many currently used expression systems containing strong but not completely repressed promoters . Such systems may be considered as constitutive ones with reduced promoter strengths, but still exhibiting all the intrinsic properties of constitutive expression systems . Although all modern expression systems are inducible, wider use of a constitutive system is evidently possible .

J Asian Nat Prod Res, 2003 Jun, 5(2), 95 - 103
Three new hecogenin glycosides from fermented leaves of Agave americana; Jin JM et al.; Eight steroidal compounds, including three new hecogenin glycosides, agamenosides D-F, were isolated from the fermented leaves of Agave americana . The structures of the new steroidal saponins were elucidated by spectroscopic data and chemical methods . The activity of the isolated compounds on deformations of mycelia germinated from conidia of Pyricularia oaryzae P-2b was evaluated.

Appl Microbiol Biotechnol, 2003 Jun, 61(5-6), 536 - 44 Epub 2003 Feb 20.
Growth and magnetosome formation by microaerophilic Magnetospirillum strains in an oxygen-controlled fermentor; Heyen U et al.; Media and growth conditions were optimized for the microaerobic cultivation of Magnetospirillum gryphiswaldense in flasks and in a fermentor, resulting in significantly increased cell and magnetosome yields, compared with earlier studies . A reliable method was established for the automatic control of low dissolved oxygen tensions (pO(2)) in the fermentor (oxystat) . Growth and magnetosome formation by M . gryphiswaldense, M . magnetotacticum and Magnetospirillum sp . AMB-1 were studied at various oxygen concentrations . Despite differences in their growth responses with respect to oxygen, we found a clear correlation between pO(2) and magnetosome formation in all three Magnetospirillum strains . Magnetite biomineralization was induced only below a threshold value of 20 mbar O(2) and optimum conditions for magnetosome formation were found at a pO(2) of 0.25 mbar (1 bar = 10(5) Pa) . A maximum yield of 6.3 mg magnetite l(-1) day(-1) was obtained with M . gryphiswaldense grown under oxystat conditions, which is the highest magnetosome productivity reported so far for a magnetotactic bacterium . In conclusion, the presented results provide the basis for large-scale cultivation of magnetospirilla under defined conditions.

Appl Microbiol Biotechnol, 2003 Jun, 61(5-6), 456 - 62 Epub 2003 Mar 18.
Optimization of the extracellular production of a bacterial phytase with Escherichia coli by using different fed-batch fermentation strategies; Kleist S et al.; The extracellular production of Escherichia coli phytase was studied in fed-batch fermentations . Two different feeding strategies were compared: control by keeping the glucose concentration constant, and control by keeping a low constant oxygen level in the medium . For the feeding control based on glucose concentration, a recently developed rapid glucose controlling system was tested for the first time in bacterial cultivations and used to establish the fermentative production of extracellular phytase with E . coli . High activity levels (120 U ml(-1)) at short cultivation times (14 h) were obtained . Even higher activity levels - albeit at longer cultivation times - were reached by applying a feeding control, the main characteristic of which was a constant low oxygen concentration . The optimum oxygen level for the production of phytase was in the range of 5-10% saturation.

Appl Microbiol Biotechnol, 2003 Jun, 61(5-6), 451 - 5 Epub 2003 Mar 06.
Nutrient regulation of epothilone biosynthesis in heterologous and native production strains; Regentin R et al.; Fermentation media with different initial concentrations of ammonium and phosphate salts were used to study the inhibitory effects of those ions on growth and production of epothilone in Sorangium cellulosum and Myxococcus xanthus . The native epothilone producer, S . cellulosum was more sensitive to ammonium and phosphate than the heterologous producer, M . xanthus . An ammonium concentration of 12 mM reduced epothilone titers by 90% in S . cellulosum but by only 40% in M . xanthus . When 5 mM phosphate was added to the medium, production in both strains was 60% lower . Higher phosphate concentrations had little additional effect on M . xanthus titers, but epothilone production with 17 mM extra-cellular phosphate in S . cellulosum was 95% lower than in the control condition . The effect of iron supplementation to the fermentation medium was also investigated . Both strains showed best production with 20 microM iron added to the medium.

J Org Chem, 2003 May 30, 68(11), 4464 - 71
A convergent solution-phase synthesis of the macrocycle Ac-Phe-{Orn-Pro-D-Cha-Trp-Arg}, a potent new antiinflammatory drug; Reid RC et al.; Relatively few cyclic peptides have reached the pharmaceutical marketplace during the past decade, most produced through fermentation rather than made synthetically . Generally, this class of compounds is synthesized for research purposes on milligram scales by solid-phase methods, but if the potential of macrocyclic peptidomimetics is to be realized, low-cost larger scale solution-phase syntheses need to be devised and optimized to provide sufficient quantities for preclinical, clinical, and commercial uses . Here, we describe a cheap, medium-scale, solution-phase synthesis of the first reported highly potent, selective, and orally active antagonist of the human C5a receptor . This compound, Ac-Phe{Orn-Pro-d-Cha-Trp-Arg}, known as 3D53, is a macrocyclic peptidomimetic of the human plasma protein C5a and displays excellent antiinflammatory activity in numerous animal models of human disease . In a convergent approach, two tripeptide fragments Ac-Phe-Orn(Boc)-Pro-OH and H-d-Cha-Trp(For)-Arg-OEt were first prepared by high-yielding solution-phase couplings using a mixed anhydride method before coupling them to give a linear hexapeptide which, after deprotection, was obtained in 38% overall yield from the commercially available amino acids . Cyclization in solution using BOP reagent gave the antagonist in 33% yield (13% overall) after HPLC purification . Significant features of the synthesis were that the Arg side chain was left unprotected throughout, the component Boc-d-Cha-OH was obtained very efficiently via hydrogenation of d-Phe with PtO(2) in TFA/water, the tripeptides were coupled at the Pro-Cha junction to minimize racemization via the oxazolone pathway, and the entire synthesis was carried out without purification of any intermediates . The target cyclic product was purified (>97%) by reversed-phase HPLC . This convergent synthesis with minimal use of protecting groups allowed batches of 50-100 g to be prepared efficiently in high yield using standard laboratory equipment . This type of procedure should be useful for making even larger quantities of this and other macrocyclic peptidomimetic drugs.

J Basic Microbiol, 2003, 43(3), 175 - 84
Distribution of zoosporic fungi in the mud of major Egyptian lakes; Ali EH et al.; Twenty-six identified and three unidentified species belonging to eight genera of zoosporic fungi were recovered from seventy-six mud samples of Egyptian lakes using sesame seeds as baits . Nineteen samples were collected along the borders of four major lakes . Pythium, Phytophthora and Allomyces, yielded the highest spectra of zoosporic species, while Aqualinderella, Blastocladia and Dictyuchus, had one species . Pythium, Aphanomyces, Allomyces, Aqualinderella, Phytophthora and Saprolegnia were the dominant zoosporic genera while Blastocladia and Dictyuchus were rare . The most prevalent species of zoosporic fungi were Aqualinderella fermentans, Aphanomyces laevis and unidentified Pythium species while the other isolated species in this study were rare . Menzalah was the richest lake with zoosporic species followed by Borolous and Karoon . Some zoosporic fungal genera were recovered from the mud of the four tested lakes (participator genera) and others from three lakes or from only two lakes . Physicochemical characteristics of the mud samples (pH, total soluble salts and organic matter) revealed no correlation with the fungal occurrence and distribution in these lakes.

Infect Immun, 2003 Jun, 71(6), 3634 - 8
Cytolethal distending toxin gene cluster in enterohemorrhagic Escherichia coli O157:H- and O157:H7: characterization and evolutionary considerations; Janka A et al.; We identified a cytolethal distending toxin (cdt) gene cluster in 87, 6, and 0% of sorbitol-fermenting (SF) enterohemorrhagic Escherichia coli (EHEC) O157:H(-), EHEC O157:H7, and E . coli O55:H7/H(-) strains, respectively . The toxin was expressed by the wild-type EHEC O157 strains and by a cdt-containing cosmid from a library of SF EHEC O157:H(-) strain 493/89 . The cdt flanks in strain 493/89 were homologous to bacteriophages P2 and lambda . Our data demonstrate that cdt, encoding a potential virulence factor, is present in the EHEC O157 complex and suggest that cdt may have been acquired by phage transduction.

J Antibiot (Tokyo), 2003 Mar, 56(3), 243 - 52
Studies on novel bacterial translocase I inhibitors, A-500359s . I . Taxonomy, fermentation, isolation, physico-chemical properties and structure elucidation of A-500359 A, C, D and G; Muramatsu Y et al.; In the course of our screening for bacterial phospho-N-acetylmuramyl-pentapeptide-translocase (translocase I: EC 2.7.8.13) inhibitors, we found inhibitory activity in the cultured broth of the strain identified as Streptomyces griseus SANK 60196 . The strain produced capuramycin and four novel capuramycin derivatives designated as A-500359 A, C, D and G . Purification and structural analysis were performed, and the structures of A-500359 A, C, D and G were elucidated as 6'''-methylcapuramycin, 3'-demethyl-6'''-methylcapuramycin, 2''-deoxy-6'''-methylcapuramycin and 3'-demethylcapuramycin, respectively.

J Antibiot (Tokyo), 2003 Mar, 56(3), 219 - 25
IB-00208, a new cytotoxic polycyclic xanthone produced by a marine-derived Actinomadura . I . Isolation of the strain, taxonomy and biological activites; Malet-Cascon L et al.; A new compound, IB-00208, has been isolated from the fermentation broth of an actinomycete isolated from a marine environment . The strain was identified as Actinomadura sp . by its chemical and phylogenetic characteristics . The compound shows cytotoxic activity on tumor cell lines and bactericidal activity against gram-positive bacteria.

J Antibiot (Tokyo), 2003 Mar, 56(3), 214 - 8
ICM0201, a new inhibitor of osteoclastogenesis from Cunninghamella sp . F-1490 . II . Structure determination and synthesis; Someno T et al.; ICM0201 (1), a new inhibitor of murine osteoclastogenesis in culture was isolated from a fermentation broth of Cunninghamella sp . F-1490 . The structure of ICM0201 was determined to be (3S,10aR)-3,4a-dihydroxy-2,3,4,4a-tetrahydro-2H-pyrano{3,2-b}benzo{e}morpholine-9-carboxylic acid by spectroscopic analyses and chemical studies . The structure of 1 is unique in that the tricycle ring system is composed of aminal and hemiacetal bonds.

J Antibiot (Tokyo), 2003 Mar, 56(3), 209 - 13
ICM0201, a new inhibitor of osteoclastogenesis from Cunninghamella sp . F-1490 . I . Taxonomy, fermentation, isolation and biological activities; Inoue H et al.; In the course of screening for inhibitors of osteoclastogenesis, a new substance designated as ICM0201 was isolated from a fermentation broth of Cunninghamella sp . F-1490 . ICM0201 inhibited the formation of osteoclasts in mouse bone marrow cells with an IC50 value of 0.78 microg/ml and showed weak cytotoxicity against bone marrow cells.

Usp Fiziol Nauk, 2003 Apr-Jun, 34(2), 21 - 32
{Recretion of enzymes and hormones by exocrine glands}; Korot'ko GF; The article reviews a poorly explored issue of secretive physiology-recretion from blood by glandulocites of various endocrinal glands of hydrolytic ferments and hormones that have been synthesized by digestive and endocrinal glands . The article discusses potential physiological role of the recretion function and the diagnostic significance of information obtained from analysis of recreted ferments and hormones in exosecretions.

FEBS Lett, 2003 May 22, 543(1-3), 174 - 8
Complementation of Escherichia coli ubiF mutation by Caenorhabditis elegans CLK-1, a product of the longevity gene of the nematode worm; Adachi A et al.; Caenorhabditis elegans CLK-1 was identified from long-lived mutant worms, and is believed to be involved in ubiquinone biosynthesis . The protein belongs to the eukaryotic CLK-1/Coq7p family, which is also similar to the bacterial Coq7 family, that hydroxylates demethoxyubiquinone, resulting in the formation of hydroxyubiquinone, a precursor of ubiquinone . In Escherichia coli, the corresponding reaction is catalyzed by UbiF, a member of a distinct class of hydroxylase . Although previous studies suggested that the eukaryotic CLK-1/Coq7 family is a hydroxylase of demethoxyubiquinone, there was no direct evidence to show the enzymatic activity of the eukaryotic CLK-1/Coq7 family . Here we show that the plasmid encoding C . elegans CLK-1 supported aerobic respiration on a non-fermentable carbon source of E . coli ubiF mutant strain and rescued the ability to synthesize ubiquinone, suggesting that the eukaryotic CLK-1/Coq7p family could function as bacterial UbiF.

Metab Eng, 2003 Jan, 5(1), 56 - 69
Metabolic engineering of beta-lactam production; Thykaer J et al.; Metabolic engineering has become a rational alternative to classical strain improvement in optimisation of beta-lactam production . In metabolic engineering directed genetic modification are introduced to improve the cellular properties of the production strains . This has resulted in substantial increases in the existing beta-lactam production processes . Furthermore, pathway extension, by heterologous expression of novel genes in well-characterised strains, has led to introduction of new fermentation processes that replace environmentally damaging chemical methods . This minireview discusses the recent developments in metabolic engineering and the applications of this approach for improving beta-lactam production.

J Epidemiol, 2003 May, 13(3), 162 - 8
Effect of diet and Helicobacter pylori infection to the risk of early gastric cancer; Lee SA et al.; BACKGROUND: The association of dietary habits and Helicobacter pylori infection with early gastric cancer is still unclear . METHODS: A hospital-based case-control study was conducted in Korea . Sixty-nine patients were newly diagnosed as having early gastric cancer at the Division of Gastroenterology, Asan Medical Center, and 199 healthy subjects who visited the Health Promotion Center of the this same hospital for annual health examinations were selected as controls . Helicobacter pylori infection status was assayed by ELISA, and information for dietary habits was obtained by interview using a semi-quantitative food frequency questionnaires . Preference for salty taste was also evaluated using a sensitive test . RESULTS: H . pylori seropositivity was observed in 88% of cases, as compared with 75% of controls (OR = 5.3, 95% confidence interval:1.7-16.5) . Adaptive salt concentration was significantly and positively associated with early gastric cancer risk (p < 0.01) . Decreased risks of early gastric cancer were observed in association with intakes of clear broth, raw vegetables, fruits, fruit or vegetable juices, and soybean curds . On the other hand, a high intake of salt-fermented fish and kimchi were associated with an elevated risk of early gastric cancer . Subjects with positive H . pylori infection and a high salty preference had a 10-fold higher risk of early gastric cancer than subjects without H . pylori infection and with a low salty preference (p for interaction = 0.047) . CONCLUSION: Some dietary factors and H . pylori infection are significantly associated with early gastric cancer . In particular, high-salty diets may enhance the effect of H . pyori infection in gastric carcinogenesis.

Proc Nutr Soc, 2003 Feb, 62(1), 201 - 6
The glycaemic index: importance of dietary fibre and other food properties; Bjorck I et al.; An increasing body of evidence suggests that a low-glycaemic-index (GI) diet has a therapeutic as well as a preventive potential in relation to the insulin resistance syndrome . The implementation of a low-GI diet, however, will require an extended list of low-GI foods to be available on the market . The tailoring of low-GI bread products offers a particular challenge due to their generally high GI and abundance in the diet . Low-GI bread products can be tailored by, for example,enclosure of cereal kernels, sour dough fermentation and/or addition of organic acids, or use of cereal genotypes with elevated contents of amylose or f-glucans . Low-GI cereal foods appear to vary in effect on 'second-meal' glucose tolerance in healthy subjects . In addition to the slow-release properties of such foods, the content of dietary fibre appears to play a role . The low glycaemia to starch in a pasta breakfast (GI 54) promoted a higher glucose tolerance and lowered triacylglycerol levels at a standardized lunch ingested 4 h later, compared with a white-wheat-bread breakfast (GI 100) . The metabolic benefits of the low GI properties per se have been demonstrated also in the longer term . Thus, a reduction in dietary GI improved glucose and lipid metabolism and normalized fibrinolytic activity in type 2 diabetics, while maintaining a similar amount and composition of dietary fibre . However, the higher dietary fibre content frequently associated with low-GI foods may add to the metabolic merits of a low-GI diet . Consequently, a low-GI barley meal rich in dietary fibre (GI 53) improved glucose tolerance from evening meal to breakfast, whereas an evening meal with pasta had no effect (GI 54) . The exchange of common high-GI bread for low-GI high-fibre bread, as the only dietary modification, improved insulin economy in women at risk of type 2 diabetes . These results are in accordance with epidemiological evidence of a reduced risk of type 2 diabetes with a low-GI diet rich in cereal fibre . It is concluded that low-GI cereal foods developed should preferably be rich in dietary fibre.

Shokuhin Eiseigaku Zasshi, 2003 Feb, 44(1), 54 - 8
{New method for the observation of gas-production using fiber-stuffed tube for coliform detection and EC-test}; Tsunoda K et al.; Gas production in lactose-containing medium (e.g., BGLB, LB, or EC medium) is the most important characteristic in the E . coli/Coliform group (C.F.G.) test on food and water . Generally a Durham tube is used as the fermentation tube, and the collected gas is analyzed . However, difficulties can arise, such as insufficient gas volume or muddy precipitates . Since air clings well to fibers in water, the Tsunoda tube (T-tube), a stalk-like tube stuffed with synthetic fibers which can capture the gas more precisely and easily, was designed . With this T-tube, air elimination during the preparation of the medium and detection of gas, even in small quantities, are both simplified . Our results show that the T-tube can replace the Durham tube as a device for detecting gas-production by the E . coli/Coliform group, with improved accuracy and sensitivity.

J Biol Chem, 2003 Aug 1, 278(31), 29009 - 15 Epub 2003 May 13.
Functional characterization and crystal structure of the C215D mutant of protein-tyrosine phosphatase-1B; Romsicki Y et al.; We have characterized the C215D active-site mutant of protein-tyrosine phosphatase-1B (PTP-1B) and solved the crystal structure of the catalytic domain of the apoenzyme to a resolution of 1.6 A . The mutant enzyme displayed maximal catalytic activity at pH approximately 4.5, which is significantly lower than the pH optimum of 6 for wild-type PTP-1B . Although both forms of the enzyme exhibited identical Km values for hydrolysis of p-nitrophenyl phosphate at pH 4.5 and 6, the kcat values of C215D were approximately 70- and approximately 7000-fold lower than those of wild-type PTP-1B, respectively . Arrhenius plots revealed that the mutant and wild-type enzymes displayed activation energies of 61 +/- 1 and 18 +/- 2 kJ/mol, respectively, at their pH optima . Unlike wild-type PTP-1B, C215D-mediated p-nitrophenyl phosphate hydrolysis was inactivated by 1,2-epoxy-3-(p-nitrophenoxy)propane, suggesting a direct involvement of Asp215 in catalysis . Increasing solvent microviscosity with sucrose (up to 40% (w/v)) caused a significant decrease in kcat/Km of the wild-type enzyme, but did not alter the catalytic efficiency of the mutant protein . Structurally, the apoenzyme was identical to wild-type PTP-1B, aside from the flexible WPD loop region, which was in both "open" and "closed" conformations . At physiological pH, the C215D mutant of PTP-1B should be an effective substrate-trapping mutant that can be used to identify cellular substrates of PTP-1B . In addition, because of its insensitivity to oxidation, this mutant may be used for screening fermentation broth and other natural products to identify inhibitors of PTP-1B.

FEMS Yeast Res, 2003 Jun, 3(4), 375 - 99
Genome-wide expression analyses: Metabolic adaptation of Saccharomyces cerevisiae to high sugar stress; Erasmus DJ et al.; The transcriptional response of laboratory strains of Saccharomyces cerevisiae to salt or sorbitol stress has been well studied . These studies have yielded valuable data on how the yeast adapts to these stress conditions . However, S . cerevisiae is a saccharophilic fungus and in its natural environment this yeast encounters high concentrations of sugars . For the production of dessert wines, the sugar concentration may be as high as 50% (w/v) . The metabolic pathways in S . cerevisiae under these fermentation conditions have not been studied and the transcriptional response of this yeast to sugar stress has not been investigated . High-density DNA microarrays showed that the transcription of 589 genes in an industrial strain of S . cerevisiae were affected more than two-fold in grape juice containing 40% (w/v) sugars (equimolar amounts of glucose and fructose) . High sugar stress up-regulated the glycolytic and pentose phosphate pathway genes . The PDC6 gene, previously thought to encode a minor isozyme of pyruvate decarboxylase, was highly induced under these conditions . Gene expression profiles indicate that the oxidative and non-oxidative branches of the pentose phosphate pathway were up-regulated and might be used to shunt more glucose-6-phosphate and fructose-6-phosphate, respectively, from the glycolytic pathway into the pentose phosphate pathway . Structural genes involved in the formation of acetic acid from acetaldehyde, and succinic acid from glutamate, were also up-regulated . Genes involved in de novo biosynthesis of purines, pyrimidines, histidine and lysine were down-regulated by sugar stress.

FEMS Yeast Res, 2003 Jun, 3(4), 341 - 6
Role of nitric oxide in the response of Saccharomyces cerevisiae cells to heat shock and high hydrostatic pressure; Domitrovic T et al.; Nitric oxide (NO) is a simple and unique molecule that has diverse functions in organisms, including intracellular and intercellular messenger . The influence of NO on cell growth of Saccharomyces cerevisiae and as a signal molecule in stress response was evaluated . Respiring cells were more sensitive to an increase in intracellular NO concentration than fermentatively growing cells . Low levels of NO demonstrated a cytoprotective effect during stress from heat-shock or high hydrostatic pressure . Induction of NO synthase was isoform-specific and dependent on the metabolic state of the cells and the stress response pathway . These results support the hypothesis that an increase in intracellular NO concentration leads to stress protection.

Mycol Res, 2003 Feb, 107(Pt 2), 223 - 30
Morphology and physiology of the dimorphic fungus Mucor circinelloides (syn . M . racemosus) during anaerobic growth; Lubbehusen TL et al.; The dimorphic Mucor circinelloides requires an anaerobic atmosphere and the presence of 30% CO2 to grow as a multipolar budding yeast, otherwise hyphal growth predominates . Establishing other means to control the morphology would be a distinct advantage in the development of a fermentation process for this organism for the production of heterologous proteins . Thus, conditions suppressing polarised growth while at the same time abolishing the CO2 requirement were investigated in submerged cultivations . It was found that supplementing cultures with mixtures of ergosterol and Tween 80 resulted in yeast-like growth under 100% N2 . Their impact on growth and morphological development was assessed at a range of concentrations . Maximum biomass levels and the specific growth rate decreased at elevated levels of ergosterol and Tween 80 . Possible effects of carbon dioxide and the added fatty acid/sterol mixture on supporting yeast growth by influencing the fluidity of the plasma membrane or affecting polarised growth are discussed.

West Afr J Med, 2002 Jul-Sep, 21(3), 244 - 7
Laboratory diagnosis of Gardnerella vaginalis vaginosis; Okwoli RN et al.; An evaluation of various laboratory detection methods and characteristics of Gardnerella vaginalis was made using high vaginal swab samples of 470 out patient clinic--attending women . Gram stain for 'clue cells' showed positive results in 118 (25.1%)cases; culture, in 100 cases, (213%) and Amine Odour (213%), in 26 cases 5.5%, Majority, 71 cases, of the culture-positive results were associated with a pH value of 6 to 7 . Gardnerella vaginalis grew predominantly in enriched culture media: Modified peptone-starch dextrose blood agar used for primary culture of organism, and also proteose peptone broth + cooked meat; Brain-heart infusion broth + 5% human serum; Brain heart infusion starch agar + 5% blood, and chocolate agar . Gardnerella vaginalis culture-positive samples also exhibited positive biochemical reactions with the hydrolysis of starch sensitivity to Bacitracin and 50 microg metronidazole, and haemolysis on human blood agar . Carbohydrate fermentation test was positive for all culture-positive cases, 100% for starch and maltose only, and negative for all the cases, 0% for Mannitol and glycerol.

Folia Microbiol (Praha), 2003, 48(1), 27 - 44
Biodegradable plastics from renewable sources; Flieger M et al.; Plastic waste disposal is a huge ecotechnological problem and one of the approaches to solving this problem is the development of biodegradable plastics . This review summarizes data on their use, biodegradability, commercial reliability and production from renewable resources . Some commercially successful biodegradable plastics are based on chemical synthesis (i.e . polyglycolic acid, polylactic acid, polycaprolactone, and polyvinyl alcohol) . Others are products of microbial fermentations (i.e . polyesters and neutral polysaccharides) or are prepared from chemically modified natural products (e.g., starch, cellulose, chitin or soy protein).

Cancer Invest, 2003 Apr, 21(2), 237 - 40
Food groups and risk of esophageal cancer in Chaoshan region of China: a high-risk area of esophageal cancer; Li K et al.; The current study was designed to investigate the role of common foods in the etiology of esophageal cancer in the Chaoshan region of China . A large case-control study was conducted to investigate 1248 patients with squamous cell carcinoma of the esophagus and 1248 controls matched by sex, age, and hospitals . After adjusting for the effects of sex, age, occupation, areas of residence, income, alcohol intake, and cigarette usage, a strong association with a clear dose-response relationship was observed between the fermented fish sauce eaten weekly and esophageal cancer (P for trend less than 0.001) . The results for pickles were similar to those for fermented fish sauce . A protective effect of fruits on esophageal cancer was observed, and this risk decreased (P for trend less than 0.001) as frequencies of fresh fruit intake increased . The risks from sowbelly and kipper were significantly higher in consumers than in nonconsumer, but the dose-response relationship was not statistically significant . No significant effects, however, were detected from vegetables, smoked foods, and so on . To the authors' knowledge, this is the first report on the relationship between fish juice and high risk of esophageal cancer in a population . Further epidemiological and experimental study are required to find a biological causal relationship between them.

Ann Nutr Metab, 2003, 47(3-4), 165 - 9
Determination of D- and L-pipecolic acid in food samples including processed foods; Fujita T et al.; BACKGROUND: Pipecolic acid, a metabolite of lysine, is found in human physiological fluids and is thought to play an important role in the central inhibitory gamma-aminobutyric acid system . However, it is unclear whether plasma D- and L-pipecolic acid originate from oral food intake or intestinal bacterial metabolites . METHODS: We analyzed the contents of D- and L-pipecolic acid in several processed foods including dairy products (cow's milk, cheese and yogurt), fermented beverages (beer and wine) and heated samples (beef, bovine liver, bread and tofu) to clarify the relationship between plasma D- and L-pipecolic acid and dietary foods . RESULTS: Our study revealed that some of the samples contained high concentrations of total pipecolic acid, and a higher proportion of L- than D-isomers . The other samples also showed high proportions of L-pipecolic acid . It was also shown that there is no significant change in the ratio of the D-isomer before and after heat treatment . The heat treatments could not cause the racemization of pipecolic acid in this study . CONCLUSION: These findings suggest that plasma pipecolic acid, particularly the D-isomer, does not originate from direct food intake and that D- and L-pipecolic acid can possibly be derived from intestinal bacterial metabolites .

Transplant Proc, 2003 May, 35(3 Suppl), 95S - 98S
Sirolimus-based immunosuppression with reduce dose cyclosporine or tacrolimus after renal transplantation; Formica RN Jr et al.; Sirolimus (SRL), a fermentation product of Streptomyces hygroscopicus, complexes with the FKBP12 to inhibit cyclin dependent kinase(s), collectively termed the target of rapamycin (TOR), causing G(1)-S phase cell cycle arrest . Safety and efficacy have been documented in clinical renal transplantation, but concerns were raised due to important biologically relevant side effects . Hyperlipidemia was identified, beginning with early clinical experiences, and the unexpected findings that SRL may exacerbate CsA associated nephrotoxicity was observed during the pivotal phase III studies . This report details results of our experience using SRL (target trough concentration, 10-15 ng/mL) with low dose CsA (target trough concentration, 50-100 ng/mL), seeking to determine whether this approach might provide effective immunosuppression while reducing associated nephrotoxicity . Among 121 renal transplant recipients, 62 received the SRL based regimen and 59 received MMF with all patients receiving CsA and prednisone . Similar to earlier clinical experiences, hematopoeitic abnormalities and hyperlipidemia were observed among patients who received SRL, and those abnormalities were readily controlled . However, unlike observations from the phase III SRL studies, renal function was not adversely affected . These findings support the growing body of evidence indicating that SRL based immunosuppression in combination low dose calcineurin inhibitors and corticosteroids is safe, efficacious, and without associated renal toxicity.

Environ Int, 2003 Aug, 29(5), 547 - 53
Investigating options for attenuating methane emission from Indian rice fields; Singh SN et al.; The development of methods and strategies to reduce the emission of methane from paddy fields is a central component of ongoing efforts to protect the Earth's atmosphere and to avert a possible climate change . It appears from this investigation that there can be more than one strategy to contain methane emission from paddy fields, which are thought to be a major source of methane emission in tropical Asia . Promising among the mitigating options may be water management, organic amendments, fertilizer application and selection of rice cultivars . It is always better to adopt multi-pronged strategies to contain CH4 efflux from rice wetlands . Use of fermented manures with low C/N ratio, application of sulfate-containing chemical fertilizers, selection of low CH4 emitting rice cultivars, and implementation of one or two short aeration periods before the heading stage can be effective options to minimize CH4 emission from paddy fields . Among these strategies, water management, which appears to be the best cost-effective and eco-friendly way for methane mitigation, is only possible when excess water is available for reflooding after short soil drying at the right timing and stage . However, in tropical Asia, rice fields are naturally flooded during the monsoonal rainy season and fully controlled drainage is often impossible . In such situation, water deficits during the vegetative and reproductive stage may drastically affect the rice yields . Thus, care must be taken to mitigate methane emission without affecting rice yields.

J Dairy Sci, 2003 Apr, 86(4), 1382 - 97
Effects of forage particle size, forage source, and grain fermentability on performance and ruminal pH in midlactation cows; Krause KM et al.; Our study investigated the effects of, and interactions between, forage particle size, level of dietary ruminally fermentable carbohydrate (RFC), and level of dietary starch on performance, chewing activity, and ruminal pH for dairy cows fed one level of dietary NDF . Twelve cows (48 DIM) were assigned to six treatments in a replicated 6 x 6 Latin square . Treatments were arranged in an incomplete 2 x 2 x 2 factorial design . Factors were: dry cracked shelled corn (DC, low RFC) or ground high-moisture corn (HMC; high RFC), finely chopped or coarse silage, and alfalfa silage as the only forage or a 50:50 ratio (DM basis) of alfalfa and corn silage . Diets combining HMC with only alfalfa silage were not included in the experiment . Diets were fed for ad libitum intake as a TMR with a concentrate:forage ratio of 61:39 . Diets based on only alfalfa silage and diets based on a mix of alfalfa and corn silage averaged 18.6 and 15.8% CP, 25.8 and 24.7% NDF, 17.7 and 14.8% ADF, and 29.1 and 37.3% starch, respectively . Mean particle sizes were 5.3, 2.7, 5.6, and 2.8 mm for coarse alfalfa, fine alfalfa, coarse corn silage, and fine corn silage, respectively . Decreasing forage particle size decreased DMI (23.3 vs . 21.6 kg) and organic matter intake (22.0 vs . 20.2 kg) . Increasing RFC decreased DMI (22.8 vs . 21.0 kg) and organic matter intake (21.5 vs . 20.0 kg) . Decreasing forage particle size increased energy-corrected milk for alfalfa based diets (34.9 vs . 37.4 kg) . Percentage of milk fat decreased with decreasing forage particle size (3.07 vs . 2.90%) and increased level of RFC (3.04 vs . 2.57%) . Percentage of protein increased when corn silage partially replaced alfalfa silage (2.84 vs . 2.90%) but decreased when HMC replaced DC (2.90 vs . 2.84%) . Apparent total tract digestibility of DM (66.7 vs . 68.5%), OM (65.9 vs . 70.7%), and starch (88.9 vs . 93.4%) increased when level of RFC was increased . Increasing level of RFC decreased mean ruminal pH from 5.82 to 5.67 and decreased minimum pH . Hours per day at which pH was <5.8, and area <5.8, increased when corn silage partially replaced alfalfa silage (2.6 vs . 4.4 h and 8.9 h x pH vs . 11.4 h x pH) and decreased further when level of RFC was increased (4.4 vs . 6.4 h and 11.4 h x pH vs . 14.3 h x pH) . Decreasing forage particle size in HMC diets increased hours and area <5.8, but for DC diets, the effect of forage particle size depended on forage source . Interactions were found between level of physically effective fiber, forage source, and level of RFC on production and pH, complicating the inclusion of these effects in dairy ration formulation and evaluation.

J Dairy Sci, 2003 Apr, 86(4), 1341 - 53
Effects of increasing levels of refined cornstarch in the diet of lactating dairy cows on performance and ruminal pH; Krause KM et al.; Our study investigated the effect of a linear increase in level of ruminally fermentable carbohydrate, at a constant level of dietary starch and fiber, on performance, microbial N yield, chewing activity, and ruminal pH of midlactation dairy cows . Eight cows (53 DIM) were assigned to four treatments in a double 4 x 4 Latin square . Diets consisted of increasing levels of refined cornstarch (0, 5.9, 11.9, and 17.9% of diet dry matter) replacing dry cracked, shelled corn so that increasing amounts of dietary starch originated from refined cornstarch . Corn gluten feed was used to balance diets for similar NDF content . The four diets averaged 17.9% CP, 27.2% NDF, 18.7% ADF, and 31.1% starch (dry matter basis) . Diets were fed for ad libitum intake and had a forage to concentrate ratio of 40:60 . Forage was coarsely chopped (13.7 mm mean particle size) alfalfa silage . Daily dry matter intake averaged 26.0 kg and tended (P = 0.08) to increase quadratically with increasing level of refined cornstarch . Milk production averaged 38.9 kg/d and milk fat percentage tended (P = 0.08) to decrease linearly, whereas percentage of protein increased quadratically, with increasing level of refined cornstarch . Yield of components and energy corrected milk was similar across diets . Total tract digestibility of starch increased linearly from 85.1% to 92.4% with increasing level of refined cornstarch . Microbial yield was unaffected by diet and averaged 371.1 g N/d . Time spent eating decreased linearly from 329 to 308 min/d when level of refined cornstarch was increased, but rumination time was unaffected . Ruminal concentration and proportion of acetate decreased linearly while concentration and proportion of propionate increased linearly with increasing level of refined cornstarch . Mean ruminal pH, time spent below pH 5.8 (h), and area below pH 5.8 (h x pH units/d) were unaffected by level of refined cornstarch and averaged 5.97, 8.4, and 2.9, respectively . Increasing the level of carbohydrates fermented in the rumen by replacing dry cracked corn with refined cornstarch (up to 57% of dietary starch) did not compromise rumen fermentation or affect performance of midlactation dairy cows.

J Dairy Sci, 2003 Apr, 86(4), 1330 - 40
Pasture intake and substitution rate effects on nutrient digestion and nitrogen metabolism during continuous culture fermentation; Bargo F et al.; A continuous culture system was used to investigate ruminal digestion in response to increased pasture intake and three different substitution rates (SR) in a 4 x 4 Latin square design . The treatments were 1) low pasture (55 g dry matter (DM)/d, 2) medium pasture (MP, 65 g DM/d), 3) high pasture (75 g DM/d), and 4) pasture (45 g DM/d) plus concentrate (PC, 30 g DM/d) . Treatments were designed to produce a low (0.33), medium (0.67), and high (1.00) SR (g of pasture/g of concentrate) by contrasting the low, medium, and high pasture intake treatments with the pasture plus concentrate treatment, respectively . Pasture was fed at 0630, 1000, 1730, and 2100 h, and concentrate at 0600 and 1700 h . Digestibility of DM and neutral detergent fiber were not affected by the amount of pasture . As the amount of pasture increased, pH decreased linearly, and total volatile fatty acid and NH3-N concentrations, and nonammonia N and bacterial N flows increased linearly . Concentrate supplementation did not affect DM digestibility at high SR but increased DM digestibility at low SR . Concentrate supplementation reduced pH and NH3-N concentrations at the three SR . Concentrate supplementation reduced the ratio of rumen degradable N to rumen degradable organic matter; however, the mechanism depended on the SR . High SR, concentrate supplementation reduced rumen degradable N, which reduced NIH-N concentration without affecting bacterial N flow . At low SR, concentrate supplementation increased rumen degradable organic matter, which reduced NH3-N concentration and increased bacterial N flow . Based on these results, at low SR, concentrate supplementation may enhance animal performance because of higher total DM intake and synthesis of microbial protein.

J Dairy Sci, 2003 Apr, 86(4), 1325 - 9
Stabilization of returned dairy products by ensiling with straw and molasses for animal feeding; Weinberg ZG et al.; Returned dairy products which are transferred to landfills might add to the environmental pollution . Such products have a high nutritional value for ruminants, but they should be stabilized to enable their use as cattle feed . The purpose of the current study was to examine stabilization of returned dairy products by ensiling in combinations with straw and molasses for animal feeding . Treatments included combinations of milk and cottage cheese with straw and molasses . Results indicate that such products ensile well with straw, and after 3 d of ensiling the pH decreased to around 4.0 . It was necessary to supplement cottage cheese with molasses, to supply a carbohydrate source for the lactic acid fermentation . The major fermentation product was lactic acid . Percentage of ammonia N (of total N) was generally higher in the silages made with cottage cheese than in those made with milk; the highest percentage (16%) was measured in the second experiment in the silages prepared with cottage cheese and straw . The study indicates the potential of stabilizing returned dairy products for animal feeding along with straw and molasses . There may also be potential for large dairy farms, or groups of smaller farms, to ensile waste milk with straw for later use as feed.

J Dairy Sci, 2003 Apr, 86(4), 1074 - 82
Low-fat set yogurt made from milk subjected to combinations of high hydrostatic pressure and thermal processing; Harte F et al.; The combined use of high hydrostatic pressure (300 to 676 MPa, 5 min) and thermal treatment (85 degrees C, 30 min) in milk for the manufacture of low-fat yogurt was studied . The objective was to reduce syneresis and improve the rheological properties of yogurt, reducing the need for thickeners and stabilizers . The use of high hydrostatic pressure alone, or after thermal treatment, reduced the lightness and increased the viscosity of skim milk . However, milk recovered its initial lightness and viscosity when thermal treatment was applied after high hydrostatic pressure . The MALDI-TOF spectra of skim milk presented monomers of whey proteins after a treatment of 676 MPa for 5 min . Yogurts made from skim milk subjected to 400 to 500 MPa and thermal treatment showed increased yield stress, resistance to normal penetration, and elastic modulus, while having reduced syneresis when compared to yogurts from thermally treated or raw milks . The combined use of thermal treatment and high hydrostatic pressure assures extensive whey protein denaturation and casein micelle disruption, respectively . Although reaggregation of casein submicelles occurs during fermentation, the net effect of the combined HHP and thermal treatment is the improvement of yogurt yield stress and reduction of syneresis.

J Dairy Sci, 2003 Apr, 86(4), 1052 - 73
Soyhulls as an alternative feed for lactating dairy cows: a review; Ipharraguerre IR et al.; Dairy producers use soyhulls, a byproduct of soybean processing, to replace either grain or forage in diets of lactating dairy cows . In view of the nutritional and economical value of soyhulls it is anticipated that this practice will continue to increase in popularity among nutritionists and producers of ruminant animals . This paper reviews information regarding the nutritional value of soyhulls and the effects of feeding this alternative feed on ruminal fermentation, nutrient digestion and utilization, and performance of dairy cows . Soyhulls can replace corn grain to supply about 30% of the dry matter (DM) in high-grain diets without negatively affecting either the fermentation or digestion of nutrients in the gastrointestinal tract or the performance of dairy cows . Additionally, data suggest that soyhulls might successfully replace forage to supply < or = 25% of the DM in diets of dairy cows when the supply of effective fiber, which includes a chemical and a physical component, remains adequate after including the hulls . However, caution should be exercised when data from different studies are extrapolated to practical situations because the response to feeding soyhulls appears to be largely affected by the type of carbohydrate being replaced by soyhulls; the amount, type, and physical form of the dietary forage; and the incidence of either negative or positive associative effects before and after the addition of soyhulls to the original diet . Unfortunately, the paucity of data from experiments in which soyhulls constituted more than 25 to 30% of the dietary DM restricts the ability to identify the maximum amount of soyhulls that can be used in diets of dairy cows . Information from studies in which > or = 25 to 30% of dietary DM supplied as either cereal grains or forages are replaced with soyhulls is needed to better understand and predict the production of dairy cows fed diets containing the hulls . This knowledge is essential for maximizing the use of soyhulls in diets for dairy cows.

Pest Manag Sci, 2003 May, 59(5), 538 - 44
Contribution of contact toxicity and wheat condition to mortality of stored-product insects exposed to spinosad; Toews MD et al.; Spinosad, a reduced-risk commercial insecticide derived from a bacterial fermentation product, possesses both contact and oral toxicities against insects . Contact toxicity of spinosad to adults of Rhyzopertha dominica (F), Sitophilus oryzae (L), and Tribolium castaneum (Herbst) was evaluated by exposure for 24 or 48 h to treated glass Petri dishes . Adults were exposed to different deposits (0.001-0.79 mg cm(-2)) of spinosad in 24-h tests and to deposits of 0, 0.0016 and 0.016mg cm(-2) in 48-h tests . Rhyzopertha dominica was most susceptible to spinosad in 24- and 48-h tests, followed by S . oryzae, and T . castaneum . The 24-h LD50 values were 0.0004, 0.077 and 0.189mg cm(-2) for R . dominica, S . oryzae, and T . castaneum, respectively . All R . dominica adults were dead following 48 h exposure to both spinosad deposits, whereas mortality of S . oryzae and T . castaneum ranged from 10 to 85% and 12 to 48%, respectively . Rhyzopertha dominica, T . castaneum, and O . surinamensis adults were exposed for 14 days to whole wheat, cracked wheat and wheat flour treated with 0, 0.1 and 1.0 mg kg(-1) of spinosad . Rhyzopertha dominica adults were highly susceptible to spinosad, followed by O . surinamensis and T . castaneum . Immatures (eggs and larvae) of T . castaneum and O . surinamensis exposed for 14 days were more susceptible on spinosad-treated whole wheat than on treated cracked wheat and wheat flour . This is the first report documenting contact activity of spinosad, and the effect of grain condition on spinosad toxicity, to stored-product insects.

Pharmacotherapy, 2003 May, 23(5), 572 - 8
Variability in polyene content and cellular toxicity among deoxycholate amphotericin B formulations; Cleary JD et al.; STUDY OBJECTIVE: To evaluate the toxicity of amphotericin B deoxycholate formulations . DESIGN: In vitro experiment . SETTING: University research center . MATERIAL: Human mononuclear THP-1 cells . INTERVENTION: The human mononuclear cells were exposed in vitro for 2 hours to the following deoxycholate formulations of amphotericin B, in 2.5- and 5-micro/ml concentrations: Apothecon, Pharmacia, Sigma, Gensia, Pharma-Tek, and VHA . MEASUREMENTS AND MAIN RESULTS: Toxicity of the amphotericin B formulations were assessed by measuring interleukin (IL)-1beta expression in an in vitro model . Amphotericin B content was measured by enzyme-linked immunosorbent assay (ELISA), and amphotericin A and B contents were assessed by spectrophotometry . Endotoxin contamination was evaluated in all reagents . Expression of IL-1beta from Sigma, Pharmacia, and Pharma-Tek formulations was increased approximately 250%, 50%, and 25%, respectively, compared with amphotericin A . Amphotericin B content of Sigma, Pharmacia, Pharma-Tek, and Gensia formulations, as measured by ELISA, was increased approximately 450%, 200%, 200%, and 100%, respectively, compared with Apothecon . This variation could not be explained by differences in amphotericin A or B content as measured by spectrophotometry . CONCLUSION: Amphotericin B is obtained from a fermentation plant and manufactured as a pharmaceutical at different facilities . Both previous clinical observations and the current in vitro evaluation revealed significant differences among the formulations . Likely, other polyenes or pyrogenic toxins in differing amounts are in these formulations, thus explaining the variability in toxicity observed among the formulations.

Biotechnol Bioeng, 2003 Jul 5, 83(1), 53 - 64
Metabolic load of recombinant protein production: inhibition of cellular capacities for glucose uptake and respiration after induction of a heterologous gene in Escherichia coli; Neubauer P et al.; The strong expression of recombinant proteins in bacteria affects the primary carbon and energy metabolism resulting in growth inhibition and acetate formation . By applying glucose pulses to fed-batch fermentations performed for production of a heterologous (alpha-glucosidase in Escherichia coli, we show that the induction of the recombinant gene strongly inhibits the maximum specific uptake capacities for glucose and the respiration capacity . The accumulation of glucose in the fermentation medium promotes the growth of plasmid-free cells . These inhibition effects are well described by including the kinetics of product formation into a recently published dynamic model (Lin et al . {2001} Biotechnol Bioeng 73:349-357) . The new model also includes the population characteristics and gives a good fit to the measured data describing growth, production, substrate consumption, by-product formation, and respiration .

Proc Nutr Soc, 2003 Feb, 62(1), 129 - 34
Why whole grains are protective: biological mechanisms; Slavin J; Epidemiological studies find that whole-grain intake is protective against cancer, cardiovascular disease, diabetes and obesity . Potential mechanisms for this protection are diverse since whole grains are rich in nutrients and phytochemicals . First, whole grains are concentrated sources of dietary fibre, resistant starch and oligosaccharides, carbohydrates that escape digestion in the small intestine and are fermented in the gut, producing short-chain fatty acids (SCFA) . SCFA lower colonic pH, serve as an energy source for the colonocytes and may alter blood lipids . These improvements in the gut environment may provide immune protection beyond the gut . Second, whole grains are rich in antioxidants, including trace minerals and phenolic compounds, and these compounds have been linked to disease prevention . Additionally, whole grains mediate insulin and glucose responses . Although lower glycaemic load and glycaemic index have been linked to diabetes and obesity, risk of cancers such as colon and breast cancer have also been linked to high intake of readily-available carbohydrate . Finally, whole grains contain many other compounds that may protect against chronic disease . These compounds include phytate, phyto-oestrogens such as lignan, plant stanols and sterols, and vitamins and minerals . As a consequence of the traditional models of conducting nutrition studies on isolated nutrients, few studies exist on the biological effects of increased whole-grain intake . The few whole-grain feeding studies that are available show improvements in biomarkers with whole-grain consumption, such as weight loss, blood lipid improvement and antioxidant protection.

Proc Nutr Soc, 2003 Feb, 62(1), 117 - 22
Process-induced changes on bioactive compounds in whole grain rye; Liukkonen KH et al.; Manufacturing of healthy wholegrain foods demands knowledge of process-induced changes in macro-, micro- and non-nutrients . The high content of dietary fibre is a challenge in relation to good product texture and sensory quality . The stability and bioavailability of bioactive compounds have a marked influence on the health effects of cereal foods . It was confirmed that sterols, folates, tocopherols and tocotrienols, alkylresorcinols, lignans, phenolic acids and total phenolics are concentrated in the bran layers of the rye grain, and are only present at low levels in the flour endosperm . The levels of folate and easily-extractable phenolic compounds increase in germination and sourdough baking, but there are negligible changes in the levels of sterols, lignans and alk(en)ylresorcinols . The levels of tocopherols and tocotrienols are reduced during the sourdough fermentation . In conclusion, many of the bioactive compounds in wholegrain rye are stable during food processing, and their levels can even be increased with suitable processing.

Proc Nutr Soc, 2003 Feb, 62(1), 107 - 15
Anti-cancer effects of butyrate: use of micro-array technology to investigate mechanisms; Williams EA et al.; Epidemiological evidence suggests that a high intake of resistant starch and NSP protects against colo-rectal cancer . The mechanisms underlying this protection are thought to be mediated by the short-chain fatty acid butyrate, which is present in the colonic lumen in millimolar concentrations as a result of bacterial fermentation of carbohydrates that have resisted digestion in the small intestine . In vitro studies have shown that butyrate displays a host of chemo-preventative properties including increased apoptosis, reduced proliferation, down regulation of angiogenesis, enhanced immunosurveillance and anti-inflammatory effects in colo-rectal cancer cell lines . However, the molecular mechanisms underlying the apparent chemo-preventative actions of butyrate are largely unknown . The evidence supporting the role of butyrate as an anti-cancer agent is reviewed, with particular emphasis on those studies that have attempted to elucidate the mechanism of action of butyrate . Our understanding of the mechanistic action of butyrate and its role in cancer prevention is likely to advance considerably in this post-genomic era with the application of genomic and proteomic technologies . Studies are described that have used gene array and proteomic techniques to investigate the response of colo-rectal cancer cells to butyrate . These pioneering studies illustrate the potential of these technologies to help characterise the molecular responses of the cancer cell to butyrate, and to define the role of butyrate (and other nutrients) in the prevention of colo-rectal cancer.

Proc Nutr Soc, 2003 Feb, 62(1), 87 - 93
Production rates and metabolism of short-chain fatty acids in the colon and whole body using stable isotopes; Pouteau E et al.; Short-chain fatty acids (SCFA; mainly acetate, propionate and butyrate) are largely produced in non-ruminants during the colonic bacterial fermentation of non-digestible carbohydrates . These intestinal exogenous SCFA pass in part through the splanchnic bed and reach the peripheral bloodstream, mixing with the endogenous circulating SCFA . The whole-body turnover of SCFA is thus composed of an endogenous peripheral turnover and an exogenous production that depends on dietary intake of non-digestible carbohydrates . In the present work methods were developed for determining the SCFA turnover in animals and in human subjects using stable isotopes . The original studies performed to determine endogenous and exogenous metabolism of SCFA in animals and in human subjects are summarised . Using intravenous infusion of 13C-labelled SCFA the whole-body turnover of acetate, propionate and butyrate was assessed in rats in a fasted state . The endogenous turnover of acetate and its oxidation were determined in healthy human subjects in the post-absorptive state, using intravenous infusion of {1-13C}acetate . Intragastric tracer infusions were performed to evaluate the splanchnic first-pass retention of acetate in adults . Finally, an original model was developed in healthy human subjects using intravenous infusion of {1-13C}acetate to determine in vivo the true colonic acetate production after ingestion of a non-digestible disaccharide . These present studies using stable isotopes provide the basis for a novel strategy to evaluate in vivo, in human subjects, the production of SCFA in the large intestine.

Proc Nutr Soc, 2003 Feb, 62(1), 81 - 6
New insight into butyrate metabolism; Bach Knudsen KE et al.; Butyrate is a C4 acid produced by microbial fermentation of carbohydrates and protein in the large intestine of all animal species . The factor of prime importance for the production rate of butyrate in the lower gut is type and levels of non-digestible carbohydrates entering the large intestine . It was previously believed that 85-90 % of the butyrate produced in the gut was cleared when passing the gut epithelium, but recent studies with catheterised pigs have shown that the concentration of butyrate in the portal vein is strongly influenced by the production rate in the large intestine . Increased gut production of butyrate further raises the circulating level of butyrate . For good reason it is not possible with current technologies to perform direct measurements of the variation in the butyrate concentration in the portal vein of human subjects, but short-chain fatty acid levels in portal blood from sudden-death victims, subjects undergoing emergency surgery or planned surgery have indicated a higher gut production and absolute and relative concentration of butyrate in non-fasted as compared with fasted human subjects . However, despite an expected higher gut production of butyrate when feeding a high-fibre rye-bread-based diet as compared with a low-fibre wheat-bread-based diet, there was no difference in absolute or relative levels of butyrate in the peripheral blood of human subjects.

Food Chem Toxicol, 2003 Jun, 41(6), 897 - 904
Safety evaluation of sources of docosahexaenoic acid and arachidonic acid for use in infant formulas in newborn piglets; Merritt RJ et al.; Human milk provides small quantities of preformed docosahexaenoic acid (DHA) and arachidonic acid (ARA), usually less than 1% of total fatty acids . Vegetable oil blends commonly used in infant formulas have, until recently, provided the essential fatty acid precursors for these long-chain polyunsaturated fatty acids (LCPUFA), but no preformed DHA and ARA . This study evaluated the safety of ingredient sources of DHA and ARA for use in infant formulas in a neonatal piglet model . Newborn piglets were allowed to suckle for 3 days and then divided into 4 feeding groups of 6 males and 6 females . Piglets were bottle-fed at frequent feeding intervals until 19 days of age . The composition of the piglet formulas was modeled after standard milk-based formulas for human infants while meeting nutritional requirements for piglets . Formulas were a control formula (no added DHA or ARA), a DHA formula providing 55 mg DHA/100 Cal, an ARA formula providing 96 mg/100 Cal ARA, and a DHA+ARA formula providing 34 mg DHA and 62 mg ARA/100 Cal . All formulas were equal in fat content and provided approximately 1000 Cal/l . The ARA-rich oil was from a fermentation product of Mortierella alpina (40 wt.% fatty acids as ARA) and DHA was from high DHA tuna oil (25 wt.% fatty acids as DHA) . There were no test article related effects of DHA and/or ARA indicative of an adverse health consequence to the animals seen in the clinical signs, body weights, food consumption, clinical chemistry, hematology, organ weights or gross or histopathology . The findings in this neonatal animal study support the safety of these ingredient oil sources of DHA and ARA for use in infant formulas.

Spectrochim Acta A Mol Biomol Spectrosc, 2003 Jun, 59(8), 1895 - 904
Observation of CO(2) in Fourier transform infrared spectral measurements of living Acholeplasma laidlawii cells; Omura Y et al.; In monitoring the time course of conformational disorder by Fourier transform infrared spectroscopy for intact Acholeplasma laidlawii cells grown at 37 degrees C on binary fatty acid mixtures containing oleic acid and for cells grown on pure palmitic acid, an absorption band at 2343 cm(-1) was observed . The band intensity was found to increase with time . This band was not observed in the spectra for isolated membranes . It is suggested that the 2343 cm(-1) band is due to CO(2) dissolved in water, most likely produced at the final point of fermentation of amino acid by this microorganism.

Trop Anim Health Prod, 2003 Apr, 35(2), 169 - 77
Energy supply to livestock from tropical rangeland during the dry season; Ahmed MM et al.; Nine key forage species (grasses and legumes), together with two types of crop residues, usually fed by farmers to their livestock, were collected from a rainfed area in western Sudan during the dry season (May-April) . The grasses investigated were Leptadena pyrotechnia, Cenchrus setigrus, Arista pallida, Eragrotis tremula, Schoenefeldia gracilis, Chloris vergata and Cenchrus biflorus . The crop residues investigated were the grasses, sorghum straw (Sorghum bichlor) and millet straw (Pennisetum typhodium) and the legumes Stylosanthes flavicans and Cajanus cajana . Estimates of organic matter (OM) degradability were done using the nylon bag technique, which was fitted into the model Y = a + b (1 - e(-ct)), in which the asymptote (a + b) represented the total potential degradability . Organic cell wall constituents and hence both metabolizable energy and total digestible energy or nutrients (TDN) were determined . S . flavicans showed the best organic matter degradability, and sorghum straw was better degraded than millet straw . The rest of the grasses showed poor OM degradability . Acid detergent insoluble nitrogen was inversely related to TDN, the latter falling within a narrow range for the different forages . Fermentable metabolizable energy differed only slightly, while the legume S . flavicans had the highest effective rumen digestible protein . Undegraded proteins were high for the straws and the grasses L . pyrotechnia and C . setigerus . Metabolizable protein and microbial protein were highest in the sorghum straw, C . setigerus and S . flavicans.

Trop Anim Health Prod, 2003 Apr, 35(2), 155 - 67
An in vitro evaluation of some unconventional ruminant feeds in terms of the organic matter digestibility, energy and microbial biomass; Al-Masri MR; In vitro organic matter apparent digestibility (IVOMAD), true digestibility (IVOMTD), metabolizable energy (ME), net energy lactation (NEL), microbial nitrogen (MN) and synthesis of microbial biomass (MBM) were estimated to predict the nutritive values of some agricultural by-products, drought-tolerant range plants and browses . The relationships between in vitro gas production (GP), and true or apparent digestibility . MN and MBM were studied utilizing an in vitro incubation technique . The values of IVOMAD, IVOMTD, ME, NEL, GP, MBM and MN varied with the studied experimental materials . The true fermentation of the outside part of Atriplex leucoclada produced a higher volume of gas than the middle or the inside parts, and this was associated with an increase in the values of IVOMAD, IVOMTD, ME and NEL . However, screening off the wood from olive cake to obtain olive cake pulp increased the IVOMAD, IVOMTD, ME, NEL and the volume of gas production from the true fermented material . One ml of gas was generated from the true degradation of 5 mg of wheat straw, Moringa oleifera, Alhagi camelorum, Eucaliptus camaldulensis and A . leucoclada, from 11 mg of Prosopsis stephaniana and olive cake pulp, and from 20 mg of olive cake or olive cake wood . The amount of MN or MBM produced from 100 mg of truly fermented organic matter depended on the kind of the fermented material and amounted to 0.7-2.9 mg or 8-34 mg, respectively . Crude fibre was negatively correlated to IVOMAD, IVOMTD, ME and NEL . Gas production was positively correlated to IVOMAD and IVOMTD but negatively correlated to MBM and MN.

Eur J Clin Microbiol Infect Dis, 2003 May, 22(5), 291 - 3 Epub 2003 May 07.
Detection of several Mycoplasma species at various anatomical sites of homosexual men; Taylor-Robinson D et al.; In order to determine the colonisation patterns of several Mycoplasma species in homosexual men, urethral, oral and rectal specimens from 10 homosexual men with acute non-gonococcal urethritis (NGU) and 18 without NGU were examined using sensitive methods . Mycoplasma hominis and Ureaplasma urealyticum existed in both groups, which is in keeping with previous studies of heterosexual men . Mycoplasma genitalium was detected in the rectum of both NGU-positive and NGU-negative men and in the urethra of one man with chlamydia-negative NGU, but not in those without urethritis . Mycoplasma fermentans was found in the throat and rectum only and Mycoplasma penetrans in all three anatomical sites . In contrast, Mycoplasma pirum was found in the rectum only, that is, in 5 of the 28 men studied . Infrequent examination of this site is a possible explanation for previous failures to detect Mycoplasma pirum at a mucosal surface.

Bioresour Technol, 2003 Mar, 87(1), 133 - 5
Calotropis procera (Sodom apple)--a potential material for enzyme purification; Kareem SO et al.; A simple method based on precipitation with Calotropis procera latex was developed for the purification of crude enzyme from fermentation broth . C . procera latex (10(-2) dilution) clarified and concentrated the crude amylase of Aspergillus oryzae 4-fold with 97% recovery of the initial amylase activity in the filtrate in a single step operation . The latex was stable at pH < or = 4.5 and there was no significant difference (P < or = 0.05) in the purification potential of the latex at 4 and 28 degrees C . This method could prove useful to developing economies of the tropics where ambient temperature is around 28 degrees C.

Curr Microbiol, 2003 Apr, 46(4), 302 - 6
Characterization of an IS element in Mycoplasma orale that is highly homologous to M . fermentans ISLE; Ditty SE et al.; In a previous study, using a primer set designed from Mycoplasma fermentans, we amplified a PCR fragment from Mycoplasma orale similar to the 206-bp DNA fragment amplified from M . fermentans insertion-sequence-like element (ISLE) . The presence of this similar ISLE fragment has the potential to cause confusion in the PCR diagnosis of M . fermentans and M . orale, which have significantly different clinical scenarios . An ISLE from three different M . orale strains was amplified by using a primer set designed from sequence within the left and right terminal stem and loop (S&L) structures flanking the ISLE of M . fermentans . Sequence analysis showed that the M . orale ISLE is 93% identical to that of M . fermentans at the nucleotide level and codes for two open reading frames also found in the M . fermentans ISLE . This is the first finding that two different mycoplasma species harbor highly homologous IS elements . This finding has great significance in clinical diagnosis and suggests a possibility of horizontal transfer of an IS element between two different mycoplasma species.

J Cell Sci, 2003 Jun 1, 116(Pt 11), 2137 - 47
Signal pathway integration in the switch from the mitotic cell cycle to meiosis in yeast; Honigberg SM et al.; Diploid yeast, like most eukaryotes, can undergo meiotic differentiation to form haploid gametes . Meiotic differentiation and cell growth (proliferation) are mutually exclusive programs, and in yeast the switch between growth and meiosis is controlled by nutritional signals . The signaling pathways that mediate nutritional controls on meiotic initiation fall into three broad classes: those that respond to nutrient starvation, those that respond to non-fermentable carbon sources, and those that respond to glucose . At the onset of meiosis, nutritional signaling pathways converge on transcriptional regulation of two genes: IME1, which encodes a transcription factor; and IME2, which encodes a protein kinase . Transcription of IME1 and IME2 trigger initiation of meiosis, and the expression of these two genes is linked with one other, with expression of later meiotic genes and with early meiotic events such as DNA replication . In addition, the signaling pathways that control IME1 and IME2 expression are themselves integrated through a variety of mechanisms . Thus the signal network that controls the switch from growth to meiotic differentiation provides a signaling code that translates different combinations of extracellular signals into appropriate cellular responses.

Antibiot Khimioter, 2002, 47(12), 3 - 5
{Mycelial wastes of penicillin fermentation as glue components}; Kadimaliev DA et al.; A new trend in the rational use of micelial wastes after antibiotics fermentation was elaborated . The modification of P . chrysogenum micelium by means of acid and alkaline solutions allows to use it as bone glue component . As a result physico-mechanical characteristics of the mixture increase, while the ratio cost-price decreases . The obtained glue composition can be used in the production of glue paper tape, for paper and wood glue.

J Am Diet Assoc, 2003 May, 103(5), 582 - 7
Kefir improves lactose digestion and tolerance in adults with lactose maldigestion; Hertzler SR et al.; OBJECTIVE: Kefir is a fermented milk beverage that contains different cultures than yogurt . The objective of this study was to determine whether kefir improves lactose digestion and tolerance in adults with lactose maldigestion . DESIGN: Randomized block design . SUBJECTS: Fifteen healthy, free-living adults with lactose maldigestion . MAIN OUTCOME MEASURES: Breath hydrogen excretion and lactose intolerance symptoms were monitored hourly for 8 hours after each test meal . INTERVENTION: Subjects were fed test meals consisting of 20 g lactose portions of milk (2% reduced fat), plain and raspberry flavored kefir, and plain and raspberry flavored yogurt, each following an overnight (12 hour) fast . STATISTICAL ANALYSIS: Mixed model ANOVA was performed on raw or transformed data, followed by Tukey HSD post hoc tests (when appropriate) . Significance was defined as P<.05 . RESULTS: The breath hydrogen area under the curve (AUC) for milk (224+/-39 ppm x h) was significantly greater than for the plain yogurt (76+/-14 ppm x h, P<.001), the plain kefir (87+/-37 ppm x h, P<.001), and the flavored yogurt (76+/-14 ppm x h, P=.005) . The flavored kefir had an intermediate response (156+/-26 ppm x h) . The yogurts and kefirs all similarly reduced the perceived severity of flatulence by 54% to 71% relative to milk . Abdominal pain and diarrhea symptoms were negligible among the five treatments . APPLICATIONS/CONCLUSION: Because kefir improved lactose digestion and tolerance in this study, its use may be another potential strategy for overcoming lactose intolerance . Further studies of other types of kefir for improving lactose digestion are warranted.

Br J Nutr, 2003 Jun, 89 Suppl 1, S39 - 43
Food sources of phyto-oestrogens and their precursors in Europe; Fletcher RJ; Phyto-oestrogens are dietary components found in some plants, which act in vivo like weak oestrogens . They may reduce the risk of some degenerative diseases moderated by oestrogen, including breast cancer and osteoporosis . The most widely studied are the isoflavones genistein and daidzein from soyabeans, but lignans may be more prevalent in the European diet . Soya foods have traditionally been consumed in the Orient for millennia, and are now widely available to European consumers . Levels of isoflavone in soyabeans from published literature vary between 560 and 3810 mg/kg, depending on variety and growing conditions . Soya protein concentrates and isolates derived from soyabeans contain 466-615 mg isoflavones/kg . Traditional soya milk, bean curds, bean sprouts, etc . contain 13 to 2030 mg isoflavone/kg, depending on the starting raw material and final water content . Fermented foods have a different isoflavone conjugate profile, which may be important in absorption and metabolism . Soya analogues of European foods include dairy and meat products, which contain 38 to 3000 mg total isoflavones/kg, depending the source of soya and dilution with other ingredients . A wide range of foods contain low levels of soya-derived isoflavones, but such foods do not make a significant contribution to mean intakes in Europe . Flaxseed is by far the richest source of lignan precursors . However, foods such as cereal brans, legumes and some vegetables are a more important source in the diets of Europeans because they are more widely consumed . For similar reasons, compared with soya isoflavones, lignans may be a more important source of phyto-oestrogens in the diets of Europeans.

Vet Pathol, 2003 May, 40(3), 340 - 2
Gastric amebiasis due to Entamoeba histolytica in a Dama wallaby (Macropus eugenii); Stedman NL et al.; A 1.5-year-old captive female Dama wallaby (Macropus eugenii) died after a 3-month period of progressive weight loss, anorexia, bloat, and diarrhea . Histopathologic examination revealed numerous Entamoeba histolytica trophozoites within the gastric mucosa and, less frequently, gastric submucosa and submucosal vessels . Immunofluorescent antibody testing confirmed the identity of the trophozoites as E . histolytica . The trophozoites were associated with mild glandular epithelial necrosis, mucosal erosions, and lymphoplasmacytic inflammation . E . histolytica most commonly causes necrotizing and ulcerative colitis in humans and captive nonhuman primates, and it causes necrotizing and ulcerative gastritis in nonhuman primates with sacculated stomachs adapted for leaf fermentation . Rare cases of gastric amebiasis also have been been reported in captive macropods, which also have complex sacculated stomachs . To our knowledge, this is the first report confirming E . histolytica as the cause of gastric amebiasis in a wallaby . The zoonotic potential of this infection in macropods is uncertain.

Biosci Biotechnol Biochem, 2003 Mar, 67(3), 475 - 9
(+)-Menthol and its hydroxy derivatives, novel fungal monoterpenols from the fusicoccin-producing fungi, Phomopsis amygdali F6a and Niigata 2; Sassa T et al.; In our search for new fusicoccins of unique diterpene glucosides from Phomopsis amygdali, we found that a fragrant substance was formed in the early stage of fusicoccin fermentation . This fragrant constituent was isolated and identified as (+)-menthol, which is a novel fungal metabolite as the enantiomer of well-known peppermint (-)-menthol . (+)-7-Hydroxymenthol and new (+)-(6S)-hydroxymenthol were also isolated and identified as fungal metabolites . In addition, p-menthanetriol, which has been reported as the first fungal monoterpene from the fungus, was also isolated . The possible biosynthetic relationship of these metabolites is discussed.

J Anim Sci, 2003 Apr, 81(4), 1040 - 50
Influence of fibrolytic enzymes on the hydrolysis and fermentation of pure cellulose and xylan by mixed ruminal microorganisms in vitro; Colombatto D et al.; A series of in vitro studies was conducted to determine the effects of adding a commercial enzyme product on the hydrolysis and fermentation of cellulose, xylan, and a mixture (1:1 wt/wt) of both . The enzyme product (Liquicell 2500, Specialty Enzymes and Biochemicals, Fresno, CA) was derived from Trichoderma reesei and contained mainly xylanase and cellulase activities . Addition of enzyme (0.5, 2.55 and 5.1 microL/g of DM) in the absence of ruminal fluid increased (P < 0.001) the release of reducing sugars from xylan and the mixture after 20 h of incubation at 20 degrees C . Incubations with ruminal fluid showed that enzyme (0.5 and 2.55 microL/g of DM) increased (P < 0.05) the initial (up to 6 h) xylanase, endoglucanase, and beta-D-glucosidase activities in the liquid fraction by an average of 85% . Xylanase and endoglucanase activities in the solid fraction also were increased (P < 0.05) by enzyme addition, indicating an increase in fibrolytic activity due to ruminal microbes . Gas production over 96 h of incubation was determined using a gas pressure measurement technique . Incremental levels of enzyme increased (P < 0.05) the rate of gas production of all substrates, suggesting that fermentation of cellulose and xylan was enzyme-limited . However, adding the enzyme at levels higher than 2.55 microL/g of DM failed to further increase the rate of gas production, indicating that the maximal level of stimulation was already achieved at lower enzyme concentrations . It was concluded that enzymes enhanced the fermentation of cellulose and xylan by a combination of pre- and postincubation effects (i.e., an increase in the release of reducing sugars during the pretreatment phase and an increase in the hydrolytic activity of the liquid and solid fractions of the ruminal fluid), which was reflected in a higher rate of fermentation.

J Anim Sci, 2003 Apr, 81(4), 843 - 55
Effect of source of energy and rate of growth on performance, carcass characteristics, ruminal fermentation, and serum glucose and insulin of early-weaned steers; Schoonmaker JP et al.; Seventy-three crossbred steers (initial BW = 170.5 +/- 5.5 kg) from The Ohio State University (Exp . 1) and 216 crossbred steers (initial BW 135.4 +/- 4.4 kg) from the University of Illinois (Exp . 2) were used to determine the effect of source of energy and rate of growth on performance, carcass characteristics, and glucose and insulin profiles on early-weaned steers . Effects of the diets used in Exp . 1 and 2 on ruminal pH and VFA concentrations were quantified using ruminally fistulated steers (Exp . 3) . Cattle were weaned at an average age of 119 d in all experiments and were allotted by age, BW, and breed to one of four diets: high-concentrate, fed ad libitum (ALCONC), high-concentrate fed to achieve a gain of either 1.2 kg/d (1.2CONC) or 0.8 kg/d (0.8CONC), or high-fiber, fed ad libitum (ALFIBER) . At 218 d of age, all steers were placed on the ALCONC diet until slaughter . Steers were implanted with Compudose at the initiation of all experiments and with Revalor-S when they were estimated to be 100 d from slaughter . When steers in Exp . 1 averaged 181 and 279 d of age, serum samples were collected to determine glucose and insulin concentrations . Steers were slaughtered when a fat thickness of 1.27 cm was reached (Exp . 1) or after 273 d on feed (Exp . 2) . In Exp . 1, days in the feedlot (P < 0.01) and age at slaughter (P < 0.01) were lowest for ALCONC and ALFIBER steers, and greatest for 0.8CONC steers . Overall, ADG was greatest for ALCONC and lowest for 0.8CONC steers; feed efficiency was lowest (P < 0.01) for ALFIBER steers . Final BW did not differ (P > 0.57) among treatments . At 181 and 218 d of age, serum insulin was increased (P < 0.10) and intramuscular fat percentage was greatest (P < 0.07), respectively, for ALCONC steers . In Exp . 2, overall ADG (P < 0.06) and final BW (P < 0.04) were greatest for ALCONC and lowest for 1.2CONC and 0.8CONC steers . Overall feed efficiency was greatest for 0.8CONC and lowest for ALFIBER (P < 0.01) . Growing phase diet did not affect marbling score at 218 d of age or at slaughter (P > 0.81) . In Exp . 3, differences in ruminal pH after feeding may have been a consequence of increasing acetate (ALFIBER), propionate (ALCONC), or a combination of VFA (0.8CONC and 1.2CONC), respectively (diet x time after feeding, P < 0.10) . Controlling growth by limit-feeding a high-concentrate diet for only 100 d does not extend the growth curve of early-weaned steers or enhance intramuscular fat deposition at slaughter compared to ad libitum intake of a high-concentrate or high-fiber diet.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 219 - 30
Composition and ethanol production potential of cotton gin residues; Agblevor FA et al.; Cotton gin residue (CGR) collected from five cotton gins was fractionated and characterized for summative composition . The major fractions of the CGR varied widely between cotton gins and consisted of clean lint (5-12%),hulls (16-48%), seeds (6-24%), motes (16-24%), and leaves (14-30%) . The summative composition varied within and between cotton gins and consisted of ash (7.9-14.6%), acid-insoluble material (18-26%), xylan (4-15%),and cellulose (20-38%) . Overlimed steam-exploded cotton gin waste was readily fermented to ethanol by Escherichia coli KO11 . Ethanol yields were feedstock and severity dependent and ranged from 58 to 92.5% of the theoretical yields . The highest ethanol yield was 191 L (50 gal)/t, and the lowest was 120 L (32 gal)/t.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 165 - 77
Effects of temperature and moisture on dilute-acid steam explosion pretreatment of corn stover and cellulase enzyme digestibility; Tucker MP et al.; Corn stover is emerging as a viable feedstock for producing bioethanol from renewable resources . Dilute-acid pretreatment of corn stover can solubilize a significant portion of the hemicellulosic component and enhance the enzymatic digestibility of the remaining cellulose for fermentation into ethanol . In this study, dilute H2SO4 pretreatment of corn stover was performed in a steam explosion reactor at 160 degrees C, 180 degrees C, and 190 degrees C, approx 1 wt % H2SO4, and 70-s to 840-s residence times . The combined severity (Log10 {Ro} - pH), an expression relating pH, temperature, and residence time of pretreatment, ranged from 1.8 to 2.4 . Soluble xylose yields varied from 63 to 77% of theoretical from pretreatments of corn stover at 160 and 180 degrees C . However, yields >90% of theoretical were found with dilute-acid pretreatments at 190 degrees C . A narrower range of higher combined severities was required for pretreatment to obtain high soluble xylose yields when the moisture content of the acidimpregnated feedstock was increased from 55 to 63 wt% . Simultaneous saccharification and fermentation (SSF) of washed solids from corn stover pretreated at 190 degrees C, using an enzyme loading of 15 filter paper units (FPU)/ g of cellulose, gave ethanol yields in excess of 85% . Similar SSF ethanol yields were found using washed solid residues from 160 and 180 degrees C pretreatments at similar combined severities but required a higher enzyme loading of approx 25 FPU/g of cellulose.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 127 - 40
Combined use of H2SO4 and SO2 impregnation for steam pretreatment of spruce in ethanol production; Soderstrom J et al.; Fuel ethanol can be produced from softwood through hydrolysis in an enzymatic process . Prior to enzymatic hydrolysis of the softwood, pretreatment is necessary . In this study, two-step steam pretreatment employing dilute H2SO4 impregnation in the first step and SO2 impregnation in the second step, to improve the overall sugar and ethanol yield, was investigated . The first pretreatment step was performed under conditions of low severity (180 degrees C, 10 min, 0.5% H2SO4) to optimize the amount of hydrolyzed hemicellulose . In the second step, the washed solid material from the first pretreatment step was impregnated with SO2 and pretreated under conditions of higher severity to make the cellulose more accessible to enzymatic attack, as well as to hydrolyze a portion of the cellulose . A wide range of conditions was used in the second step to determine the most favorable combination . The temperatures investigated were between 190 and 230 degrees C, the residence times were 2, 5, and 10 min; and the SO2 concentration was 3% . The effect of pretreatment was assessed by both enzymatic hydrolysis of the solids and by simultaneous saccharification and fermentation (SSF) of the whole slurry, after the second pretreatment step . For each set of pretreatment conditions, the liquid fraction was also fermented to determine any inhibitory effects . Ethanol yield using the SSF configuration reached 66% of the theoretical value for pretreatment conditions in the second step of 210 degrees C and 5 min . The sugar yield using the separate hydrolysis and fermentation configuration reached 71% for pretreatment conditions of 220 degrees C and 5 min.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 87 - 100
Hydrothermal pretreatment conditions to enhance ethanol production from poplar biomass; Negro MJ et al.; Pretreatment has been recognized as a key step in enzyme-based conversion processes of lignocellulose biomass to ethanol . The aim of this study is to evaluate two hydrothermal pretreatments (steam explosion and liquid hot water) to enhance ethanol production from poplar (Populus nigra) biomass by a simultaneous saccharification and fermentation (SSF) process . The composition of liquid and solid fractions obtained after pretreatment, enzymatic digestibility, and ethanol production of poplar biomass pretreated at different experimental conditions was analyzed . The best results were obtained in steam explosion pretreatment at 210 C and 4 min, taking into account cellulose recovery above 95%, enzymatic hydrolysis yield of about 60%, SSF yield of 60% of theoretical, and 41% xylose recovery in the liquid fraction . Large particles can be used for poplar biomass in both pretreatments, since no significant effect of particle size on enzymatic hydrolysis and SSF was obtained.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 69 - 85
Dilute-sulfuric acid pretreatment of corn stover in pilot-scale reactor: investigation of yields, kinetics, and enzymatic digestibilities of solids; Schell DJ et al.; Corn stover is a domestic feedstock that has potential to produce significant quantities of fuel ethanol and other bioenergy and biobased products . However, comprehensive yield and carbon mass balance information and validated kinetic models for dilute-sulfuric acid (H2SO4) pretreatment of corn stover have not been available . This has hindered the estimation of process economics and also limited the ability to perform technoeconomic modeling to guide research . To better characterize pretreatment and assess its kinetics, we pretreated corn stover in a continuous 1 t/d reactor . Corn stover was pretreated at 20% (w/w) solids concentration over a range of conditions encompassing residence times of 3-12 min, temperatures of 165- 195 degrees C, and H2SO4 concentrations of 0.5-1.4% (w/w) . Xylan conversion yield and carbon mass balance data were collected at each run condition . Performance results were used to estimate kinetic model parameters assuming biphasic hemicellulose hydrolysis and a hydrolysis mechanism incorporating formation of intermediate xylo-oligomers . In addition, some of the pretreated solids were tested in a simultaneous saccharification and fermentation (SSF) process to measure the reactivity of their cellulose component to enzymatic digestion by cellulase enzymes . Monomeric xylose yields of 69-71% and total xylose yields (monomers and oligomers) of 70-77% were achieved with performance level depending on pretreatment severity . Cellulose conversion yields in SSF of 80-87% were obtained for some of the most digestible pretreated solids.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 403 - 12
Fungi allergens produced by solid-state fermentation process: optimization and allergen characterization; Hasan SD et al.; Allergenic extracts were produced from Drechslera (Helminthosporium) monoceras biomass cultured by solid-state fermentation using wheat bran as the substrate . The main fermentation variables were selected by statistical design, and the optimized biomass yield (1.43 mg/{g of dry substrate d}) was obtained at pH 9.5 and 45.8% moisture . The allergenic extracts were produced from crude extract by protein precipitation and polyphenol removal . Proteins in the range of 16-160 kDa were identified in the extracts . Their reactions in patients were characterized by in vivo cutaneous tests (positive in 40% of the atopic patients) and by dot-blotting assays.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 319 - 35
Optimization of SO2-catalyzed steam pretreatment of corn fiber for ethanol production; Bura R et al.; A batch reactor was employed to steam explode corn fiber at various degrees of severity to evaluate the potential of using this feedstock as part of an enzymatically mediated cellulose-to-ethanol process . Severity was controlled by altering temperature (150-230 degrees C), residence time (1-9 min), and SO2 concentration (0-6% {w/w} dry matter) . The effects of varying the different parameters were assessed by response surface modeling . The results indicated that maximum sugar yields (hemicellulose-derived water soluble, and cellulose-derived following enzymatic hydrolysis) were recovered from corn fiber pretreated at 190 degrees C for 5 minutes after exposure to 3% SO2 . Sequential SO2-catalyzed steam explosion and enzymatic hydrolysis resulted in a conversion efficiency of 81% of the combined original hemicellulose and cellulose in the corn fiber to monomeric sugars . An additional posthydrolysis step performed on water soluble hemicellulose stream increased the concentration of sugars available for fermentation by 10%, resulting in the high conversion efficiency of 91% . Saccharomyces cerevisiae was able to ferment the resultant corn fiber hydrolysates, perhydrolysate, and liquid fraction from the posthydrolysis steps to 89, 94, and 85% of theoretical ethanol conversion, respectively . It was apparent that all of the parameters investigated during the steam explosion pretreatment had a significant effect on sugar recovery, inhibitory formation, enzymatic conversion efficiency, and fermentation capacity of the yeast.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 615 - 28
Limits for alkaline detoxification of dilute-acid lignocellulose hydrolysates; Nilvebrant NO et al.; In addition to fermentable sugars, dilute-acid hydrolysates of lignocellulose contain compounds that inhibit fermenting microorganisms, such as Saccharomyces cerevisiae . Previous results show that phenolic compounds and furan aldehydes, and to some extent aliphatic acids, act as inhibitors during fermentation of dilute-acid hydrolysates of spruce . Treatment of lignocellulose hydrolysates with alkali, usually in the form of overliming to pH 10.0, has been frequently employed as a detoxification method to improve fermentability . A spruce dilute-acid hydrolysate was treated with NaOH in a factorial design experiment, in which the pH was varied between 9.0 and 12.0, the temperature between 5 and 80 degrees C, and the time between 1 and 7 h . Already at pH 9.0, >25% of the glucose was lost when the hydrolysate was treated at 80 degrees C for 1 h . Among the monosaccharides, xylose was degraded faster under alkaline conditions than the hexoses (glucose, mannose, and galactose), which, in turn, were degraded faster than arabinose . The results suggest that alkali treatment of hydrolysates can be performed at temperatures below 30 degrees C at any pH between 9.0 and 12.0 without problems with sugar degradation or formation of inhibiting aliphatic acids . Treatment with Ca(OH)2 instead of NaOH resulted in more substantial degradation of sugars . Under the harsher conditions of the factorial design experiment, the concentrations of furfural and 5-hydroxymethylfurfural decreased while the total phenolic content increased . The latter phenomenon was tentatively attributed to fragmentation of soluble aromatic oligomers in the hydrolysate . Separate phenolic compounds were affected in different ways by the alkaline conditions with some compounds showing an increase in concentration while others decreased . In conclusion, the conditions used for detoxification with alkali should be carefully controlled to optimize the positive effects and minimize the degradation of fermentable sugars.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 523 - 46
Conversion of sugarcane bagasse to carboxylic acids using a mixed culture of mesophilic microorganisms; Thanakoses P et al.; Using the MixAlco process, biomass can be converted into carboxylic acids, which can be chemically converted into mixed alcohol fuels . This study focused on the use of countercurrent fermentation to anaerobically convert sugarcane bagasse and chicken manure to mixed carboxylic acids using a mixed culture of mesophilic microorganisms from terrestrial and marine sources . Bagasse was pretreated with lime to increase digestibility . The continuum particle distribution model (CPDM) simulated continuous fermentors based on data collected from batch experiments . This model saves considerable time in determining optimum operating conditions . For an 80% bagasse/20% chicken manure fermentation with terrestrial inoculum at a volatile solids loading rate (VSLR) of 7.36 g/(L of liquid d) and a liquid residence time (LRT) of 8.88 d, total carboxylic acid productivity, total acid selectivity, and yield were 2.49 g/(L of liquid d), 0.581 g of total acid/ g of VS digested, and 0.338 g of total acid/g of VS fed, respectively, at a concentration of 18.7 g of total acid/L . At the same VSLR and LRT, fermentation with marine inoculum gave higher total acid productivity, total acid selectivity, and yield than fermentation with terrestrial inoculum . For an 80% bagasse/20% chicken manure fermentation with marine inoculum at a VSLR of 3.83 g/(L of liquid d) and an LRT of 12.1 d, total carboxylic acid productivity, total acid selectivity, and yield were 1.38 g/(L of liquid d), 0.667 g of total acid/g of VS digested, and 0.359 g of total acid/g of VS fed, respectively, at a concentration of 16.2 g of total acid/L.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 843 - 51
Ester fuels and chemicals from biomass; Olson ES et al.; Bench-scale research demonstrated that using an efficient esterification step to integrate an ethanol with a carboxylic acid fermentation stream offers potential for producing valuable ester feedstocks and fuels . Polar organic acids from bacterial fermentations are difficult to extract and purify, but formation of the ammonium salts and their conversion to esters facilitates the purifications . An improved esterification procedure gave high yields of esters, and this method will lower the cost of ester production . Fuel characteristics have been determined for a number of ester-gasoline blends with promising results for lowering Reid vapor pressure and raising octane numbers.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 737 - 48
Xylanase production by Penicillium canescens 10-10c in solid-state fermentation; Bakri Y et al.; Filamentous fungi have been widely used to produce hydrolytic enzymes for industrial applications, including xylanases, whose levels in fungi are generally much higher than those in yeast and bacteria . We evaluated the influence of carbon sources, nitrogen sources, and moisture content on xylanase production by Penicillium canescens 10-10c in solid-state fermentation . Among agricultural wastes tested (wheat bran, untreated wheat straw, treated wheat straw, beet pulp, and soja meal), untreated wheat straw gave the highest production of xylanase . Optimal initial moisture content for xylanase production was 83% . The addition of 0.4 g of xylan or easily metabolizable sugar, such as glucose and xylose, at a concentration of 2 % to wheat straw enhanced xylanase production . In solid-state fermentation, even at high concentrations of glucose or xylose (10%), catabolic repression was minimized compared to the effect observed in liquid culture . Yeast extract was the best nitrogen source among the nitrogen sources investigated: peptone, ammonium nitrate, sodium nitrate, ammonium chloride, and ammonium sulfate . A combination of yeast extract and peptone as nitrogen sources led to the best xylanase production.

Appl Biochem Biotechnol, 2003 Spring, 105 -108, 715 - 24
Xylanase production by Trichoderma reesei rut C-30 on rice straw; Colina A et al.; Xylanase production of Trichoderma reesei Rut C-30 was examined at different initial pH values (4.8, 5.9, and 7.0) on rice straw in shake flasks, and in a fermentor, for the best pH condition . Enzyme performance was tested on ammonia-treated dwarf elephant grass . The maximum xylanase activities, 92 and 122 IU/mL, were obtained at pH 4.8 in the shake flasks and fermentor, respectively, in which good growth of the fungus was observed during the first 24 h and consumption of proteins dissolved from the rice straw caused the pH to rise later to values between 6.4 and 6.7 (optimal for xylanase production) . The xylanases from T . reesei were as effective as Multifect XL, a commercial enzyme preparation, in hydrolyzing ammonia-treated elephant grass.

J Agric Food Chem, 2003 May 7, 51(10), 3060 - 6
Volatile compounds of wines produced by cells immobilized on grape skins; Mallouchos A et al.; A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae cells on grape skins . Repeated batch fermentations were conducted using the immobilized biocatalyst as well as the free yeast cells at 25, 20, 15, and 10 degrees C . The major volatile byproducts were determined by GC, whereas the minor volatile constituents were extracted in dichloromethane and analyzed by HRGC-MS . The qualitative profiles of the wines produced were similar in every case . Immobilized cells gave wines with higher contents of ethyl and acetate esters that increased with temperature decreases from 25 to 15 degrees C . The amount of volatile alcohols was more pronounced in wines produced by free cells and decreased dramatically at low fermentation temperatures (10 degrees C).

Int J Biol Macromol, 2003 Mar, 32(1-2), 28 - 35
Tensile deformation of bacterial cellulose composites; Astley OM et al.; The polymeric basis for the mechanical properties of primary plant cell walls has been investigated by forming analogous composites based on fermentation of the bacterium Acetobacter xylinus, either alone or in the presence of xyloglucan or pectin . Simultaneous small-angle X-ray scattering and uniaxial deformation experiments has shown how the cellulose microfibrils reorient during deformation . Despite very different stress/strain curves, the reorientation behaviour is similar, regardless of the presence or absence of xyloglucan or pectin . A simple theory has been developed to predict the orientation behaviour . This is qualitatively similar to the measured behaviour, but differs quantitatively.

Asian Pac J Cancer Prev, 2002, 3(4), 345 - 352
Gastric Cancer: the Roles of Diet, Alcohol Drinking, Smoking and Helicobacter pylori in Northeastern Thailand; Sriamporn S et al.; The incidence of gastric cancer in the countries of South East Asia is variable, ranging from age-standardized rates of 20.9/105 (men) and 10.4/105 (women) in Hanoi, Vietnam to 4.1/105 (men) and 2.1/105 (women) in Khon Kaen, Thailand . The reasons for these differences are unknown . Possible explanations are differences in dietary habits, alcohol drinking, smoking and/or the prevalence of infection with Helicobacter pylori (H . pylori) . A case-control study was conducted in Khon Kaen, Thailand, to study the role of these factors in gastric cancer carcinogenesis . 131 gastric cancer cases and 262 matched controls were recruited for the study . Information on dietary habits, alcohol drinking and smoking were collected by a structured questionnaire . Blood samples were available from 111 cases and 232 controls for H . pylori assay . Using an unconditional logistic regression model controlling for age and sex, we assessed the effects of dietary habits, alcohol drinking, smoking and H . pylori infection on the risk of gastric cancer . A high intake of salt (OR=1.8; 95%CI 1.1-3.0) and fermented foods (OR=1.9; 95%CI 1.1-3.3) was found to be associated with an increased risk . Preference for spicy food was not associated with gastric cancer risk in this population . Although there were negative associations between gastric cancer and vegetable and fruit intake, they were rather weak (OR 0.8 for both) and non significant . There were also weak (non-significant) associations with smoking and alcohol consumption, and no association with H . pylori infection (OR=0.6; 95%CI 0.4-1.0) . Infection of H . pylori was associated with various indicators of crowding.

J Antibiot (Tokyo), 2003 Feb, 56(2), 87 - 90
A new IL-1 receptor inhibitor 139A: fermentation, isolation, physico-chemical properties and structure; Jing C et al.; Interleukin (IL)-1 is known to be a cytokine which plays a major role in pathological conditions like septic shock, inflammation and auto-immune disease, hence, methods that reduce the activity of IL-1 have an impact on clinical medicine . Inhibiting the binding of IL-1 to IL-1 receptors is one of the methods . A new inhibitor of IL-1 receptor, 139A, was isolated from the fermentation broth of Streptomyces sp.139 . It was extracted from the broth filtrate, purified by Diaion HP-20, cation exchange resin and DEAE Sephadex A-25 . 139A was identified as polysaccharide, its structure was elucidated on the basis of spectral analysis, the immobilized ligand IL-1 receptor binding assay (IL-ILRBA) proved 139A can competitively inhibits the binding of IL-1 to IL-1 receptors.

J Antibiot (Tokyo), 2003 Feb, 56(2), 80 - 6
FR235222, a fungal metabolite, is a novel immunosuppressant that inhibits mammalian histone deacetylase (HDAC) II . Biological activities in animal models; Mori H et al.; FR235222, a novel immunosuppressant which possesses potent inhibitory effects on the activity of mammalian histone deacetylases (HDACs), has been isolated from the fermentation broth of a fungus, Acremonium sp . No . 27082 . FR235222 exhibited marked immunosuppressive effects on mouse ex vivo splenic T-lymphocyte proliferation, mouse delayed type hypersensitivity (DTH) response, rat adjuvant-induced arthritis (AA) and rat heterotopic cardiac transplantation . These results showed potential clinical use of this compound as a new type immunosuppressant in the fields of autoimmune diseases and organ transplantations.

J Antibiot (Tokyo), 2003 Feb, 56(2), 72 - 9
FR235222, a fungal metabolite, is a novel immunosuppressant that inhibits mammalian histone deacetylase (HDAC) . I . Taxonomy, fermentation, isolation and biological activities; Mori H et al.; A cyclic tetrapeptide FR235222, a novel immunosuppressant, has been isolated from the fermentation broth of a fungus, Acremonium sp . No . 27082 . FR235222 showed potent and selective inhibitory effects on both T cell proliferation and lymphokine production . Further study has revealed this compound exhibits potent inhibitory effects on the activity of mammalian histone deacetylases (HDACs).

J Ind Microbiol Biotechnol, 2003 Mar, 30(3), 183 - 9 Epub 2003 Feb 27.
Optimization of phytase production by solid substrate fermentation; Bogar B et al.; The production of phytase by three feed-grade filamentous fungi ( Aspergillus ficuum NRRL 3135, Mucor racemosus NRRL 1994 and Rhizopus oligosporus NRRL 5905) on four commonly used natural feed ingredients (canola meal, cracked corn, soybean meal, wheat bran) was studied in solid substrate fermentation (SSF) . A . ficuum NRRL 3135 had the highest yield {15 IU phytase activity/g dry matter (DM)} on wheat bran . By optimizing the supplementation of wheat bran with starch and (NH(4))(2)SO(4), phytase production increased to 25 IU/g DM . Optimization was carried out by Plackett-Burman and central composite experimental designs . Using optimized medium, phytase, phosphatase, alpha-amylase and xylanase production by A . ficuum NRRL 3135 was studied in Erlenmeyer flask and tray SSF . By scaling up SSF from flasks to stationary trays, activities of 20 IU phytase activity/g DM were reproducibly obtained.

J Ind Microbiol Biotechnol, 2003 Mar, 30(3), 175 - 82 Epub 2003 Feb 22.
Degradation of DNA during the autolysis of Saccharomyces cerevisiae; Zhao J et al.; The autolysis of yeast cells has practical implications in the production of fermented foods and beverages and flavourants for food processing . Protein and RNA degradation during yeast autolysis are well described but the fate of DNA is unclear . Yeast cells ( Saccharomyces cerevisiae) were autolysed by incubating suspensions at 30-60 degrees C (pH 7.0), and at pH 4.0-7.0 (40 degrees C) for 10-14 days . Up to 55% of total DNA was degraded, with consequent leakage into the extracellular environment of mainly 3'- and 5'-deoxyribonucleotides, and lesser amounts of polynucleotides . The rate and extent of DNA degradation, composition of the DNA degradation products and DNase activity were affected by temperature and pH . The highest amount of DNA degradation occurred at 40 degrees C and pH 7.0, where the highest DNase activity was recorded . DNase activity was lowest at 60 degrees C and pH 4.0, where the proportion of polynucleotides in the degradation products was higher.

J Ind Microbiol Biotechnol, 2003 Mar, 30(3), 161 - 7 Epub 2003 Mar 01.
Approaches to stabilization of inter-domain recombination in polyketide synthase gene expression plasmids; Hu Z et al.; Regions of extremely high sequence identity are recurrent in modular polyketide synthase (PKS) genes . Such sequences are potentially detrimental to the stability of PKS expression plasmids used in the combinatorial biosynthesis of polyketide metabolites . We present two different solutions for circumventing intra-plasmid recombination within the megalomicin PKS genes in Streptomyces coelicolor . In one example, a synthetic gene was used in which the codon usage was reengineered without affecting the primary amino acid sequence . The other approach utilized a heterologous subunit complementation strategy to replace one of the problematic regions . Both methods resulted in PKS complexes capable of 6-deoxyerythronolide B analogue biosynthesis in S . coelicolor CH999, permitting reproducible scale-up to at least 5-l stirred-tank fermentation and a comparison of diketide precursor incorporation efficiencies between the erythromycin and megalomicin PKSs.

J Ind Microbiol Biotechnol, 2003 Mar, 30(3), 157 - 60 Epub 2003 Mar 04.
Production of laccase and manganese peroxidase by Fomes sclerodermeus grown on wheat bran; Papinutti VL et al.; The aim of this work was to study the growth and production of ligninolytic enzymes by Fomes sclerodermeus using a natural medium based on wheat bran as the principal substrate in a solid-state fermentation . Growth was monitored by measuring the chitin content in the substrate . The maximum rate of growth was observed between days 7 and 18 . A 38% total dry-weight loss of the substrate was measured after 28 days of cultivation . Differential hydrolysis of the substrate revealed that cellulose was more extensively degraded than lignin . In the 28-day incubation period, the losses of cellulose and lignin were 38 and 15%, respectively . No lignin peroxidase activity was found in any of the media tested . The maximum manganese-dependent peroxidase activity recorded was 6.3 U g(-1) at 14 days, while the maximum laccase activity was 270 U g(-1) at 28 days post-inoculation . Addition of commonly used inducers such as copper or manganese did not produce a further increase in the enzyme activities, nor did addition of glucose, asparagine, or malt extract.

Curr Genet, 2003 Jun, 43(3), 139 - 60 Epub 2003 Apr 25.
Transcriptional control of nonfermentative metabolism in the yeast Saccharomyces cerevisiae; Schuller HJ; Although sugars are clearly the preferred carbon sources of the yeast Saccharomyces cerevisiae, nonfermentable substrates such as ethanol, glycerol, lactate, acetate or oleate can also be used for the generation of energy and cellular biomass . Several regulatory networks of glucose repression (carbon catabolite repression) are involved in the coordinate biosynthesis of enzymes required for the utilization of nonfermentable substrates . Positively and negatively acting complexes of pleiotropic regulatory proteins have been characterized . The Snf1 (Cat1) protein kinase complex, together with its regulatory subunit Snf4 (Cat3) and alternative beta-subunits Sip1, Sip2 or Gal83, plays an outstanding role for the derepression of structural genes which are repressed in the presence of a high glucose concentration . One molecular function of the Snf1 complex is deactivation by phosphorylation of the general glucose repressor Mig1 . In addition to regulation of alternative sugar fermentation, Mig1 also influences activators of respiration and gluconeogenesis, although to a lesser extent . Snf1 is also required for conversion of specific regulatory factors into transcriptional activators . This review summarizes regulatory cis-acting elements of structural genes of the nonfermentative metabolism, together with the corresponding DNA-binding proteins (Hap2-5, Rtg1-3, Cat8, Sip4, Adr1, Oaf1, Pip2), and describes the molecular interactions among general regulators and pathway-specific factors . In addition to the influence of the carbon source at the transcriptional level, mechanisms of post-transcriptional control such as glucose-regulated stability of mRNA are also discussed briefly.

Biosens Bioelectron, 2003 Jul, 18(7), 857 - 65
High-stability non-invasive autoclavable naked optical CO2 sensor; Ge X et al.; The fabrication and characterization of a high-stability non-invasive autoclavable naked optical CO(2) sensor is described in this report . The sensor was made by using 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS) as the fluorescence dye and cetyltrimethylammonium hydroxide (CTMAOH) as the phase transfer agent (the base) . A highly hydrophobic two-component silicone film was used as the polymer matrix, which overcame some of the limitations of the existing plastic type CO(2) sensors, such as dye leaching and cross-sensitivity to ions . To improve the stability of the sensor, several affecting factors were investigated . Experimental results showed that sufficient base and a small amount of water in the sensing film were critical factors that affected the stability of the sensor . Although the sensor was more stable when kept in water, the function of the sensor could recover when the sensor kept in air was transferred into water . The sensor has a lifetime of several months . The detection limit of the sensing film was about 0.03% . The average response and recovery times were 0.66 and 1.94 min, respectively . It had no cross-sensitivity to salt concentrations in the range of 0-0.2 M and to pH in the range of 5.6-8.0, so it can be used in processes with changing ion concentration and pH . It was sterilizable and could be autoclaved many times without losing its sensitivity . The applicability of the sensor in real application was successfully tested in the fermentation of Escherichia coli.

J Nat Prod, 2003 Apr, 66(4), 551 - 3
Isolation, structure, and HIV-1 integrase inhibitory activity of Cytosporic acid, a fungal metabolite produced by a Cytospora sp; Jayasuriya H et al.; HIV-1 integrase is a critical enzyme for replication of HIV, and its inhibition has the potential to lead to an anti-retroviral therapy that has advantages over existing therapies . Cytosporic acid (1) is a polyketide-derived novel natural product that was isolated from a fermentation broth of the filamentous fungus Cytospora sp . collected from Puerto Rico . It inhibited strand transfer reaction of HIV-1 integrase with an IC(50) of 20 microM . The isolation, structure elucidation, relative stereochemistry, and activity of 1 are described.

J Nat Prod, 2003 Apr, 66(4), 511 - 4
A new phytotoxic nonenolide from Phoma herbarum; Rivero-Cruz JF et al.; Reinvestigation of the fermentation broth and mycelium of the fungus Phoma herbarum led to the isolation of a new phytotoxic nonenolide, namely, (7R,9R)-7-hydroxy-9-propyl-5-nonen-9-olide, which was designated with the trivial name herbarumin III (3) . The known compounds herbarumins I (1) and II (2) were also obtained . The structure of 3 was elucidated by spectroscopic methods and molecular modeling . Compounds 1-3 interacted with bovine-brain calmodulin and inhibited the activation of the calmodulin-dependent enzyme cAMP phosphodiesterase.

Biodegradation, 2002, 13(6), 411 - 24
Complete dechlorination of tetrachloroethene to ethene in presence of methanogenesis and acetogenesis by an anaerobic sediment microcosm; Aulenta F et al.; An anaerobic consortium taken from brackish sediments, enriched by PCE/CH3OH sequential feeding, was capable of completely dechlorinating tetrachloroethene (PCE) to ethene (ETH) . In batch experiments, PCE (0.5 mM) was dechlorinated to ethene (ETH) in approximately 75 h with either CH3OH or H2 as the electron donor . When VC (0.5 mM) was added instead of PCE it was dechlorinated without any initial lag by the PCE/CH3OH enriched consortium, although at a lower dechlorination rate . In batch tests H2 could readily replace CH3OH for supporting PCE dechlorination, with a similar PCE dechlorination rate and product distribution with respect to those observed with methanol . This indicates that H2 production during CH3OH fermentation was not the rate-limiting step of PCE or VC dechlorination . Acetogenesis was the predominant activity when methanol was present . A remarkable homoacetogenic activity was also observed when hydrogen was supplied instead of methanol.

Oecologia, 2003 Jun, 136(1), 14 - 27 Epub 2003 Apr 24.
The maximum attainable body size of herbivorous mammals: morphophysiological constraints on foregut, and adaptations of hindgut fermenters; Clauss M et al.; An oft-cited nutritional advantage of large body size is that larger animals have lower relative energy requirements and that, due to their increased gastrointestinal tract (GIT) capacity, they achieve longer ingesta passage rates, which allows them to use forage of lower quality . However, the fermentation of plant material cannot be optimized endlessly; there is a time when plant fibre is totally fermented, and another when energy losses due to methanogenic bacteria become punitive . Therefore, very large herbivores would need to evolve adaptations for a comparative acceleration of ingesta passage . To our knowledge, this phenomenon has not been emphasized in the literature to date . We propose that, among the extant herbivores, elephants, with their comparatively fast passage rate and low digestibility coefficients, are indicators of a trend that allowed even larger hindgut fermenting mammals to exist . The limited existing anatomical data on large hindgut fermenters suggests that both a relative shortening of the GIT, an increase in GIT diameter, and a reduced caecum might contribute to relatively faster ingesta passage; however, more anatomical data is needed to verify these hypotheses . The digestive physiology of large foregut fermenters presents a unique problem: ruminant-and nonruminant-forestomachs were designed to delay ingesta passage, and they limit food intake as a side effect . Therefore, with increasing body size and increasing absolute energy requirements, their relative capacity has to increase in order to compensate for this intake limitation . It seems that the foregut fermenting ungulates did not evolve species in which the intake-limiting effect of the foregut could be reduced, e.g . by special bypass structures, and hence this digestive model imposed an intrinsic body size limit . This limit will be lower the more the natural diet enhances the ingesta retention and hence the intake-limiting effect . Therefore, due to the mechanical characteristics of grass, grazing ruminants cannot become as big as the largest browsing ruminant . Ruminants are not absent from the very large body size classes because their digestive physiology offers no particular advantage, but because their digestive physiology itself intrinsically imposes a body size limit . We suggest that the decreasing ability for colonic water absorption in large grazing ruminants and the largest extant foregut fermenter, the hippopotamus, are an indication of this limit, and are the outcome of the competition of organs for the available space within the abdominal cavity . Our hypotheses are supported by the fossil record on extinct ruminant/tylopod species which did not, with the possible exception of the Sivatheriinae, surpass extant species in maximum body size . In contrast to foregut fermentation, the GIT design of hindgut fermenters allows adaptations for relative passage acceleration, which explains why very large extinct mammalian herbivores are thought to have been hindgut fermenters.

Diabetologia, 2003 May, 46(5), 659 - 65 Epub 2003 Apr 24.
Prior short-term consumption of resistant starch enhances postprandial insulin sensitivity in healthy subjects; Robertson MD et al.; AIMS/HYPOTHESIS: Diets rich in insoluble-fibre are linked to a reduced risk of both diabetes and cardiovascular disease; however, the mechanism of action remains unclear . The aim of this study was to assess whether acute changes in the insoluble-fibre (resistant starch) content of the diet would have effects on postprandial carbohydrate and lipid handling . METHODS: Ten healthy subjects consumed two identical, low-residue diets on separate occasions for 24 h (33% fat; <2 g dietary fibre) . Of the diets one was supplemented with 60 g resistant starch (Novelose 260) . On the following morning a fibre-free meal tolerance test (MTT) was carried out (59 g carbohydrate; 21 g fat; 2.1 kJ) and postprandial insulin sensitivity (SI(ORAL)) assessed using a minimal model approach . RESULTS: Prior resistant starch consumption led to lower postprandial plasma glucose (p=0.037) and insulin (p=0.038) with a higher insulin sensitivity(44+/-7.5 vs 26+/-3.5 x 10(-4) dl kg(-1) min(-1) per micro Uml(-1); p=0.028) and C-peptide-to-insulin molar ratio (18.7+/-6.5 vs 9.7+/-0.69; p=0.017) . There was no effect of resistant starch consumption on plasma triacylglycerol although non-esterified fatty acid and 3-hydroxybutyrate levels were suppressed 5 h after the meal tolerance test . CONCLUSION : Prior acute consumption of a high-dose of resistant starch enhanced carbohydrate handling in the postprandial period the following day potentially due to the increased rate of colonic fermentation.

J Exp Bot, 2003 May, 54(386), 1421 - 9
Genetic and biochemical analysis of anaerobically-induced enzymes during seed germination of Echinochloa crus-galli varieties tolerant and intolerant of anoxia; Fukao T et al.; To compare the regulation of anaerobic metabolism during germination in anoxia-tolerant and intolerant plants, enzymes associated with anaerobic metabolism such as sucrose synthase, aldolase, enolase, pyruvate decarboxylase (PDC), alcohol dehydrogenase (ADH), and aldehyde dehydrogenase (ALDH) were assayed in two varieties of Echinochloa crus-galli, formosensis (tolerant) and praticola (intolerant) . The initial and intervening enzymes of the pathway (sucrose synthase and aldolase) and enzymes in the last part of the pathway (PDC, ADH and ALDH) revealed similar changing patterns in activities during germination . This implies that each group of enzymes may be controlled by an identical regulatory mechanism . During anoxia, activities of all enzymes increased 1.5-30-fold in both varieties compared to their activities under aerobic conditions . Activities of sucrose synthase, enolase and ADH exhibited the same induction patterns under anoxia in formosensis and praticola . However, the activities of aldolase, ALDH and PDC were more strongly induced in formosensis under anoxia (1.2-2-fold) than in praticola . These enzymes were also assayed in F(3) families which varied in their anaerobic germinability . For PDC, activities under anoxia in anoxia-tolerant families were similar to those of an anoxia-intolerant family during the whole period although the family did not exhibit anaerobic germinability . This suggests that there is no correlation between PDC activity and anaerobic germinability . For ALDH, activities were more strongly induced under anoxia in anoxia-tolerant families than in anoxia-intolerant families, a trend also exhibited by the parents . This indicates that ALDH may play a role in detoxifying acetaldehyde formed through alcoholic fermentation during anaerobic germination.

Huan Jing Ke Xue, 2003 Jan, 24(1), 70 - 4
{Comparison and analysis of hydrogen production capacity with different acidogenic fermentative microflora}; Ren N et al.; The biogas(hydrogen) production capacity of ethanol type and propionic acid type fermentative microflora were compared, and the movement of hydrogen production rate according to the succession of microflora from propionic acid type to ethanol type was investigated . It was demonstrated that under the same loading rate, microflora of ethanol type fermentation had relatively high hydrogen production rate and specific hydrogen production rate, maximum hydrogen production rate was 14.99 L/d, maximum specific hydrogen production rate was 3586.45 mmol/(kg.d) . While, for the microflora of propionic acid type fermentation, hydrogen production rate and specific hydrogen production rate were extremely low, maximum hydrogen production rate was 3.62 L/d, maximum specific hydrogen production rate was 196.46 mmol/(kg.d) . In order to obtain high yield of H2 in the fermentation process, ethanol type fermentation was preferred and propionic and type fermentation had to be avoided.

Carbohydr Res, 2003 May 1, 338(10), 1127 - 32
Specificity of yeast (Saccharomyces cerevisiae) in removing carbohydrates by fermentation; Yoon SH et al.; The specificity of Saccharomyces cerevisiae yeast on the removal of carbohydrates by fermentation was studied . The common monosaccharides, D-glucose, D-fructose, D-mannose, and D-galactose were completely removed; D-glucuronic acid and D-ribose were partially removed; but D-xylose, D-rhamnose, and L-sorbose were not removed and were completely resistant . Of four glycosides, methyl and phenyl alpha- and beta-D-glucopyranosides, three of the four were partially removed and methyl beta-D-glucopyranoside was not removed . The disaccharides, maltose, sucrose, and turanose were completely removed, while cellobiose, lactose, and melibiose were completely resistant . Isomaltose and alpha,alpha-trehalose were partially removed . Maltotriose and raffinose were partially removed, but isomaltotriose and melezitose were completely resistant . The tetrasaccharides, maltotetraose, isomaltotetraose, and acarbose, were completely resistant . Further, the yeast enzymes did not alter any of the resistant carbohydrates by transglycosylation or condensation reactions or by any other types of reactions.

J Pharmacol Exp Ther, 2003 May, 305(2), 557 - 64
Cytochrome P450 2E1 (CYP2E1) is the principal enzyme responsible for urethane metabolism: comparative studies using CYP2E1-null and wild-type mice; Hoffler U et al.; Urethane ({carbonyl-(14)C}ethyl carbamate) is a fermentation by-product in alcoholic beverages and foods and is classified as reasonably anticipated to be a human carcinogen . Early studies indicated that while CYP2E1 is involved, esterases are the primary enzymes responsible for urethane metabolism . Using CYP2E1-null (KO) mice, current studies were undertaken to elucidate CYP2E1's contribution to urethane metabolism . {Carbonyl-(14)C}urethane was administered by gavage to male CYP2E1-null and wild-type mice at 10 or 100 mg/kg and its metabolism and disposition were investigated . CO(2) was confirmed as the main metabolite of urethane . Significant inhibition of urethane metabolism to CO(2) occurred in CYP2E1-null versus wild-type mice . Pharmacokinetic modeling of (14)CO(2) exhalation data revealed that CYP2E1 is responsible for approximately 96% of urethane metabolism to CO(2) in wild-type mice . The contributions of other enzymes to urethane metabolism merely account for the remaining 4% . The half-life of urethane in wild-type and CYP2E1-null mice was estimated at 0.8 and 22 h, respectively . Additionally, the concentration of urethane-derived radioactivity in blood and tissues was dose-dependent and significantly higher in CYP2E1-null mice . High-performance liquid chromatography analysis showed only urethane in the plasma and liver extracts of CYP2E1-null mice . Because the lack of CYP2E1 did not completely inhibit urethane metabolism, the disposition of 10 mg/kg urethane was compared in mice pretreated with the P450 inhibitor, 1-aminobenzotriazole or the esterase inhibitor, paraoxon . Unlike paraoxon, 1-aminobenzotriazole resulted in significant inhibition of urethane metabolism to CO(2) in both genotypes . In conclusion, this work demonstrated that CYP2E1, not esterase, is the principal enzyme responsible for urethane metabolism.

J Dairy Sci, 2003 Mar, 86(3), 926 - 33
Effect of storage method on fermentation characteristics, aerobic stability, and forage intake of tropical grasses ensiled in round bales; Gonzalez G et al.; The effect of storage method and length of the fermentation period were evaluated to determine fermentation characteristics, aerobic stability, and forage intake of native tropical grasses ensiled in round bales . Bales were stored under shade or under direct sunlight and sampled after seven periods of ensiling (0, 1, 4, 11, 25, 53, and 111 d) . For the aerobic stability and nutritive value studies, three bales were opened from each storage method after 53 and 111 d, exposed to air, and fed to heifers for voluntary consumption . The storage method did not affect fermentation characteristics, however, during aerobic exposure, silage storage under shade resulted in less deterioration as evidenced by lower pH, temperature, and microbial populations . Also, the shorter length of fermentation (53 d) resulted in lower pH and temperature and enhanced aerobic stability . The combination of silage storage under shade and short length of fermentation resulted in the most stable silages and did not improve the nutritive value of native tropical grasses ensiled in large round bales as evidenced by similar forage intake.

J Dairy Sci, 2003 Mar, 86(3), 916 - 25
Effects of transition diets varying in dietary energy density on lactation performance and ruminal parameters of dairy cows; Rabelo E et al.; Forty cows and twenty heifers were used to study the effects of dietary energy density during late gestation and early lactation on lactation performance and ruminal parameters . A 2 x 2 factorial arrangement of treatments was used . During prepartum (-28 d to calving), animals were fed a low energy density diet {DL; 1.58 Mcal of net energy for lactation (NE(L))/kg, 40% neutral detergent fiber (NDF) and 38% nonfiber carbohydrate (NFC)} or a high energy diet (DH; 1.70 Mcal NE(L)/kg, 32% NDF and 44% NFC) . After calving, half of the cows from each prepartum treatment group were assigned to a low energy density diet (L; 1.57 Mcal NE(L)/kg, 30% NDF and 41% NFC) or a high energy density diet (H; 1.63 Mcal NE(L)/kg, 25% NDF and 47% NFC) until d 20 postpartum . After d 20, all cows were fed H until d 70 . Animals fed DH had 19.8% greater dry matter intake (DMI; % of body weight) and 21.5% greater energy intake than animals fed DL prepartum and the response was greater for cows compared to heifers . Animals fed DH had lower ruminal pH compared to animals fed DL, but no major changes in volatile fatty acid concentrations were observed . Effects of dietary energy density during prepartum on postpartum production responses were dependent on parity . Primiparous cows fed DL had higher 3.5% fat-corrected milk yield and milk fat production and percentage during the first 10 wk of lactation than those fed DH . Prepartum diet did not affect lactation performance of multiparous cows . Cows fed H had higher DMI and energy intake for the first 20 d of lactation compared to cows fed L . Diets did not affect DMI after the third wk of lactation . Milk production increased faster for cows fed H compared to cows fed L . Animals fed DL-L sequence of treatments tended to have the lowest energy intake during the first 10 wk of lactation . Prepartum treatments did not affect ruminal fermentation characteristics postpartum . Cows fed H had lower ruminal pH and higher propionate concentrations than cows fed L . No prepartum x postpartum interactions were observed for ruminal fermentation parameters . The effects of DH on prepartum DMI did not carry over to the postpartum period or influence early postpartum production . Increasing concentrate content of the diet immediately postpartum instead of delaying the increase until d 21 postpartum is associated with a higher rate of increase.in milk production and higher DMI.

FEMS Yeast Res, 2001 Apr, 1(1), 73 - 8
Kinetics of active alpha-glucoside transport in Saccharomyces cerevisiae; Stambuk BU et al.; alpha-Glucosides are the most abundant fermentable sugars in the industrial applications of Saccharomyces cerevisiae, and the active transport across the plasma membrane is the rate-limiting step for their metabolism . In this report we performed a detailed kinetic analysis of the active alpha-glucoside transport system(s) present in a wild-type strain, and in strains with defined alpha-glucoside permeases . Our results indicate that the wild-type strain harbors active transporters with high and low affinity for maltose and trehalose, and low-affinity transport systems for maltotriose and alpha-methylglucoside . The maltose permease encoded by the MAL21 gene showed a high affinity (K(m) approximately 5 mM) for maltose, and a low affinity (K(m) approximately 90 mM) for trehalose . On the other hand, the alpha-glucoside permease encoded by the AGT1 gene had a high affinity (K(m) approximately 7 mM) for trehalose, a low affinity (K(m) approximately 18 mM) for maltose and maltotriose, and a very low affinity (K(m) approximately 35 mM) for alpha-methylglucoside.

FEMS Yeast Res, 2003 Apr, 3(2), 223 - 32
The ALEU2 gene--a new component for an Arxula adeninivorans-based expression platform; Wartmann T et al.; The ALEU2 gene, encoding beta-isopropylmalate dehydrogenase, was isolated from the non-conventional yeast Arxula adeninivorans . The isolated gene harbours an open reading frame of 1086 bp, encoding a putative protein of 362 amino acids . The derived protein sequence shares a high degree of homology with other fungal beta-isopropylmalate dehydrogenases thus confirming the identity of the gene . The isolated ALEU2 gene was tested for its suitability to complement the auxotrophy of an A . adeninivorans aleu2 host . For this purpose the plasmid pAL-ALEU2m which contains the ALEU2 gene as a selection marker and the 25S rDNA for targeting was employed in transformation experiments . Transformants harboured a single copy of the heterologous DNA and were found to be mitotically stable . For assessment of heterologous gene expression, two model genes were incorporated into the vector: the GFP gene, encoding intracellular green fluorescent protein, and the HSA gene, encoding the secreted human serum albumin . For expression control, both gene sequences were fused to the constitutive A . adeninivorans-derived TEF1 promoter and the Saccharomyces cerevisiae-derived PHO5 terminator . In the respective recombinant strains the GFP was localised in the cytoplasm, whereas more than 95% of the HSA accumulated in the culture medium . In initial fermentation trials using a 200-ml shake flask, maximal HSA product levels were observed after 96 h of cultivation.

FEMS Yeast Res, 2003 Apr, 3(2), 185 - 9
Production of ethanol from L-arabinose by Saccharomyces cerevisiae containing a fungal L-arabinose pathway; Richard P et al.; The fungal pathway for L-arabinose catabolism converts L-arabinose to D-xylulose 5-phosphate in five steps . The intermediates are, in this order: L-arabinitol, L-xylulose, xylitol and D-xylulose . Only some of the genes for the corresponding enzymes were known . We have recently identified the two missing genes for L-arabinitol 4-dehydrogenase and L-xylulose reductase and shown that overexpression of all the genes of the pathway in Saccharomyces cerevisiae enables growth on L-arabinose . Under anaerobic conditions ethanol is produced from L-arabinose, but at a very low rate . The reasons for the low rate of L-arabinose fermentation are discussed.

FEMS Yeast Res, 2003 Apr, 3(2), 149 - 57
The impact of brewing yeast cell age on fermentation performance, attenuation and flocculation; Powell CD et al.; Individual cells of the yeast Saccharomyces cerevisiae exhibit a finite replicative lifespan, which is widely believed to be a function of the number of divisions undertaken . As a consequence of ageing, yeast cells undergo constant modifications in terms of physiology, morphology and gene expression . Such characteristics play an important role in the performance of yeast during alcoholic beverage production, influencing sugar uptake, alcohol and flavour production and also the flocculation properties of the yeast strain . However, although yeast fermentation performance is strongly influenced by the condition of the yeast culture employed, until recently cell age has not been considered to be important to the process . In order to ascertain the effect of replicative cell age on fermentation performance, age synchronised populations of a lager strain were prepared using sedimentation through sucrose gradients . Each age fraction was analysed for the ability to utilise fermentable sugars and the capacity to flocculate . In addition cell wall properties associated with flocculation were determined for cells within each age fraction . Aged cells were observed to ferment more efficiently and at a higher rate than mixed aged or virgin cell cultures . Additionally, the flocculation potential and cell surface hydrophobicity of cells was observed to increase in conjunction with cell age . The mechanism of ageing and senescence in brewing yeast is a complex process, however here we demonstrate the impact of yeast cell ageing on fermentation performance.

FEMS Yeast Res, 2001 Jul, 1(2), 139 - 49
Modulating the distribution of fluxes among respiration and fermentation by overexpression of HAP4 in Saccharomyces cerevisiae; van Maris AJ et al.; The tendency of Saccharomyces cerevisiae to favor alcoholic fermentation over respiration is a complication in aerobic, biomass-directed applications of this yeast . Overproduction of Hap4p, a positive transcriptional regulator of genes involved in respiratory metabolism, has been reported to positively affect the balance between respiration and fermentation in aerobic glucose-grown batch cultures . In this study, the effects of HAP4 overexpression have been quantified in the prototrophic S . cerevisiae strain CEN.PK 113-7D under a variety of growth conditions . In aerobic glucose-limited chemostat cultures, overexpression of HAP4 increased the specific growth rate at which aerobic fermentation set in by about 10% relative to the isogenic wild-type . Upon relief of glucose-limited conditions, the HAP4-overexpressing strain produced slightly less ethanol than the wild-type strain . The effect of Hap4p overproduction was most drastic in aerobic, glucose-grown chemostat cultures in which ammonium was limiting . In such cultures, the biomass yield on glucose was double that of the wild-type.

FEMS Yeast Res, 2001 Jul, 1(2), 111 - 25
Functional genomic analysis of a commercial wine strain of Saccharomyces cerevisiae under differing nitrogen conditions; Backhus LE et al.; DNA microarray analysis was used to profile gene expression in a commercial isolate of Saccharomyces cerevisiae grown in a synthetic grape juice medium under conditions mimicking a natural environment for yeast: High-sugar and variable nitrogen conditions . The high nitrogen condition displayed elevated levels of expression of genes involved in biosynthesis of macromolecular precursors across the time course as compared to low-nitrogen . In contrast, expression of genes involved in translation and oxidative carbon metabolism were increased in the low-nitrogen condition, suggesting that respiration is more nitrogen-conserving than fermentation . Several genes under glucose repression control were induced in low-nitrogen in spite of very high (17%) external glucose concentrations, but there was no general relief of glucose repression . Expression of many stress response genes was elevated in stationary phase . Some of these genes were expressed regardless of the nitrogen concentration while others were found at higher levels only under high nitrogen conditions . A few genes, FSP2, RGS2, AQY1, YFL030W, were expressed more strongly with nitrogen limitation as compared to other conditions.

FEMS Yeast Res, 2002 May, 2(2), 245 - 8
Aspergillus oryzae in solid-state and submerged fermentations . Progress report on a multi-disciplinary project; te Biesebeke R et al.; We report the progress of a multi-disciplinary research project on solid-state fermentation (SSF) of the filamentous fungus Aspergillus oryzae . The molecular and physiological aspects of the fungus in submerged fermentation (SmF) and SSF are compared and we observe a number of differences correlated with the different growth conditions . First, the aerial hyphae which occur only in SSFs are mainly responsible for oxygen uptake . Second, SSF is characterised by gradients in temperature, water activity and nutrient concentration, and inside the hyphae different polyols are accumulating . Third, pelleted growth in SmF and mycelial growth in SSF show different gene expression and protein secretion patterns . With this approach we aim to expand our knowledge of mechanisms of fungal growth on solid substrates and to exploit the biotechnological applications.

FEMS Yeast Res, 2002 May, 2(2), 225 - 32
Genetic engineering of brewing yeast to reduce the content of ethanol in beer; Nevoigt E et al.; The GPD1 gene encoding the glycerol-3-phosphate dehydrogenase was overexpressed in an industrial lager brewing yeast (Saccharomyces cerevisiae ssp . carlsbergensis) to reduce the content of ethanol in beer . The amount of glycerol produced by the GPD1-overexpressing yeast in fermentation experiments simulating brewing conditions was increased 5.6 times and ethanol was decreased by 18% when compared to the wild-type . Overexpression of GPD1 does not affect the consumption of wort sugars . Only minor changes in the concentration of higher alcohols, esters and fatty acids could be observed in beer produced by the GPD1-overexpressing brewing yeast . However, the concentrations of several other by-products, particularly acetoin, diacetyl and acetaldehyde, were considerably increased.

FEMS Yeast Res, 2002 May, 2(2), 137 - 49
The level of MXR1 gene expression in brewing yeast during beer fermentation is a major determinant for the concentration of dimethyl sulfide in beer; Hansen J et al.; DMS (dimethyl sulfide) is an important beer flavor compound which is derived either from the beer wort production process or via the brewing yeast metabolism . We investigated the contribution of yeast MXR1 gene activity to the final beer DMS content . The MXR1-CA gene from Saccharomyces carlsbergensis (synonym of Saccharomyces pastorianus) lager brewing yeast was isolated and sequenced, and found to be 88% identical with Saccharomyces cerevisiae MXR1 . Inactive deletion alleles of both genes were substituted for their functional counterparts in S . carlsbergensis . Such yeasts fermented well and did not form DMS from dimethyl sulfoxide . Overexpression in brewing yeast of MXR1 from non-native promoters with various strengths and transcription profiles resulted in an enhanced and correlated DMS production . The promoters of MXR1 and MXR1-CA contain conserved Met31p/Met32p binding sites, and in accordance with this were found to be co-regulated with the genes of the sulfur assimilation pathway . In addition, conserved YRE-like DNA sequences are present in these promoters, indicating that Yap1p may also take part in the control of these genes.

FEMS Yeast Res, 2002 May, 2(2), 93 - 102
Evidence for involvement of Saccharomyces cerevisiae protein kinase C in glucose induction of HXT genes and derepression of SUC2; Brandao RL et al.; The PKC1 gene in the yeast Saccharomyces cerevisiae encodes protein kinase C that is known to control a mitogen-activated protein (MAP) kinase cascade consisting of Bck1, Mkk1 and Mkk2, and Mpk1 . This cascade affects the cell wall integrity but the phenotype of Pkc1 mutants suggests additional targets which have not yet been identified . We show that a pkc1Delta mutant, as opposed to mutants in the MAP kinase cascade, displays two major defects in the control of carbon metabolism . It shows a delay in the initiation of fermentation upon addition of glucose and a defect in derepression of SUC2 gene after exhaustion of glucose from the medium . After addition of glucose the production of both ethanol and glycerol started very slowly . The V(max) of glucose transport dropped considerably and Northern blot analysis showed that induction of the HXT1, HXT2 and HXT4 genes was strongly reduced . Growth of the pkc1Delta mutant was absent on glycerol and poor on galactose and raffinose . Oxygen uptake was barely present . Derepression of invertase activity and SUC2 transcription upon transfer of cells from glucose to raffinose was deficient in the pkc1Delta mutant as opposed to the wild-type . Our results suggest an involvement of Pkc1p in the control of carbon metabolism which is not shared by the downstream MAP kinase cascade.

FEMS Yeast Res, 2002 Aug, 2(3), 403 - 7
Oenological properties of Hanseniaspora osmophila and Kloeckera corticis from wines produced by spontaneous fermentations of normal and dried grapes; Granchi L et al.; Strains of Hanseniaspora osmophila and Kloeckera corticis, isolated from wines produced by spontaneous fermentations of normal and dried grapes, were characterized for their fermentation behavior with and without SO(2) at 25 degrees C . All isolates behaved as glucophilic yeasts and yielded ethanol at concentrations of about 9% (v/v); acetic acid, acetaldehyde, ethyl acetate and acetoin were always produced to high concentrations . SO(2) addition had no significant effect on growth yield and fermentation rate . These metabolic features were maintained in the presence of 400 g l(-1) of sugars and at 15 degrees C, and were quite similar to those shown by Saccharomycodes ludwigii . Therefore, H . osmophila and K . corticis should be considered detrimental yeast species, particularly in fermentations of musts from dried grapes.

FEMS Yeast Res, 2002 Aug, 2(3), 363 - 9
High-level production and secretion of recombinant proteins by the dimorphic yeast Arxula adeninivorans; Wartmann T et al.; The non-conventional dimorphic thermo- and salt-resistant yeast Arxula adeninivorans has been developed as a host for heterologous gene expression . For assessment of the system two model genes have been selected: the GFP gene encoding the intracellular green fluorescent protein, and the HSA gene encoding the secreted human serum albumin . The expression system includes two host strains, namely A . adeninivorans LS3, which forms budding cells at 30 degrees C and mycelia at >42 degrees C, and the strain A . adeninivorans 135, which forms mycelia at temperatures as low as 30 degrees C . For expression control the constitutive A . adeninivorans-derived TEF1-promoter and S . cerevisiae-derived PHO5-terminator were selected . The basic A . adeninivorans transformation/expression vector pAL-HPH1 is further equipped with the Escherichia coli-derived hph gene conferring hygromycin B resistance and the 25S rDNA from A . adeninivorans for rDNA targeting . Transformants were obtained for both budding cells and mycelia . In both cell types similar expression levels were achieved and the GFP was localised in the cytoplasm while more than 95% of the HSA accumulated in the culture medium . In initial fermentation trials on a 200-ml shake flask scale maximal HSA product levels were observed after 96 h of cultivation.

FEMS Yeast Res, 2002 Aug, 2(3), 277 - 82
The non-oxidative pentose phosphate pathway controls the fermentation rate of xylulose but not of xylose in Saccharomyces cerevisiae TMB3001; Johansson B et al.; Saccharomyces cerevisiae is able to ferment xylose, when engineered with the enzymes xylose reductase (XYL1) and xylitol dehydrogenase (XYL2) . However, xylose fermentation is one to two orders of magnitude slower than glucose fermentation . S . cerevisiae has been proposed to have an insufficient capacity of the non-oxidative pentose phosphate pathway (PPP) for rapid xylose fermentation . Strains overproducing the non-oxidative PPP enzymes ribulose 5-phosphate epimerase (EC 5.1.3.1), ribose 5-phosphate ketol isomerase (EC 5.3.1.6), transaldolase (EC 2.2.1.2) and transketolase (EC 2.2.1.1), as well as all four enzymes simultaneously, were compared with respect to xylose and xylulose fermentation with their xylose-fermenting predecessor S . cerevisiae TMB3001, expressing XYL1, XYL2 and only overexpressing XKS1 (xylulokinase) . The level of overproduction in S . cerevisiae TMB3026, overproducing all four non-oxidative PPP enzymes, ranged between 4 and 23 times the level in TMB3001 . Overproduction of the non-oxidative PPP enzymes did not influence the xylose fermentation rate in either batch cultures of 50 g l(-1) xylose or chemostat cultures of 20 g l(-1) glucose and 20 g l(-1) xylose . The low specific growth rate on xylose was also unaffected . The results suggest that neither of the non-oxidative PPP enzymes has any significant control of the xylose fermentation rate in S . cerevisiae TMB3001 . However, the specific growth rate on xylulose increased from 0.02-0.03 for TMB3001 to 0.12 for the strain overproducing only transaldolase (TAL1) and to 0.23 for TMB3026, suggesting that overproducing all four enzymes has a synergistic effect . TMB3026 consumed xylulose about two times faster than TMB30001 in batch culture of 50 g l(-1) xylulose . The results indicate that growth on xylulose and the xylulose fermentation rate are partly controlled by the non-oxidative PPP, whereas control of the xylose fermentation rate is situated upstream of xylulokinase, in xylose transport, in xylose reductase, and/or in the xylitol dehydrogenase.

FEMS Yeast Res, 2002 Dec, 2(4), 563 - 73
The dynamics of the Saccharomyces carlsbergensis brewing yeast transcriptome during a production-scale lager beer fermentation; Olesen K et al.; The transcriptome of a lager brewing yeast (Saccharomyces carlsbergensis, syn . of S . pastorianus), was analysed at 12 different time points spanning a production-scale lager beer fermentation . Generally, the average expression rapidly increased and had a maximum value on day 2, then decreased as the sugar got consumed . Especially genes involved in protein and lipid biosynthesis or glycolysis were highly expressed during the beginning of the fermentation . Similarities as well as significant differences in expression profiles could be observed when comparing to a previous transcriptome analysis of a laboratory yeast grown in YPD . The regional distribution of various expression levels on the chromosomes appeared to be random or near-random and no reduction in expression near telomeres was observed.

FEMS Yeast Res, 2003 Mar, 3(1), 11 - 6
A flow-cytometric method for determination of yeast viability and cell number in a brewery; Boyd AR et al.; A flow-cytometric assay, using the fluorescent dye, oxonol, for the simultaneous determination of yeast cell viability and cell number is described . The assay was optimised, and trialed at a brewery for 6 months . The flow-cytometry assay offered a substantially reduced error in viability determination, compared to methylene blue which is the industry standard for measuring viability . Further, by calculating yeast cell number at the same time, this assay provides a reliable method for determining pitching rate, allowing increased quality control of subsequent fermentations.

Hiroshima J Med Sci, 2003 Mar, 52(1), 9 - 13
Inhibition by long-term fermented miso of induction of pulmonary adenocarcinoma by diisopropanolnitrosamine in Wistar rats; Shiraki K et al.; The present study was designed to investigate the effects of fermented miso in the diet on the development of lung tumors initiated by diisopropanolnitrosamine (BHP) in male Slc:Wistar rats . A total of 63 animals, 6 weeks of age, were divided into 4 groups and given BHP (2000 ppm) in their drinking water for 10 weeks . After the carcinogen treatment the rats were fed a normal control MF solid diet, or the same diet containing 10% long-term or short-term fermented miso for 12 weeks . The long-term fermented miso significantly reduced the number of lung tumors, adenocarcinomas and PCNA strongly positive tumors as compared with the short-term fermented miso . The present results thus indicate that dietary supplementation with long-term fermented miso could exert chemopreventive effects on lung carcinogenesis.

J Epidemiol, 2003 Jan, 13(1 Suppl), S13 - 22
Self-administered food frequency questionnaire used in the 5-year follow-up survey of the JPHC Study: questionnaire structure, computation algorithms, and area-based mean intake; Sasaki S et al.; In this section we described the structure of the self-administered semiquantitative food frequency questionnaire used in the 5-year follow-up survey of the JPHC study, the computation algorithms, and the area-based mean intakes of nutrients and food groups in the subjects of the validation study . The FFQ consists of five sections: 1) semiquantitative frequency questions for rice and miso (fermented soybean paste)-soup, 2) those for alcoholic beverages, 3) those for vitamin supplements, 4) those for foods and beverages, and 5) questions on dietary and cooking behaviors . From the questions, intakes of nutrients and foods by food groups were computed . Although most of them were computed from the frequency and relative portion size indicated in the replies, together with the fixed portion size, a seasonal coefficient was added in the computation of vegetables and fruits . Only frequency of intake and fixed portion size were used for computation of beverages . Sugar and cream added in coffee and tea were computed from the frequency of coffee and tea intake . The intakes of cooking oil, cooking salt (sodium), and salt in noodle-soup were estimated from the questions of relative preference of oil, salt, and noodle-soup.

Dent Clin North Am, 2003 Apr, 47(2), 319 - 36
Nutrition and dental caries; Mobley CC; Promotion of sound dietary practices is an essential component of caries management, along with fluoride exposure and oral hygiene practices . Scientific discoveries have lead to better understanding of the caries process, the ever-expanding food supply, and the interaction between the two . Fermentable carbohydrates interact dynamically with oral bacteria and saliva, and these foods will continue to be a major part of a healthful diet . Dental health professionals can serve their patients and the public by providing comprehensive oral health care and by promoting lifestyle behaviors to improve oral and general health within the time constraints of their practice . Dietary advice given should not contradict general health principles when providing practical guidance to reduce caries risk . The following principles should guide messages: * Encourage balanced diets based on moderation and variety as depicted by the Food Guide Pyramid and the Dietary Guidelines for Americans to provide a sound approach . Avoid references to "bad" foods and focus on "good" diets that include a variety of foods . * Give examples of how combining and sequencing foods can enhance mastication, saliva production, and oral clearance at each eating occasion . Combining dairy foods with sugary foods, raw foods with cooked, and protein-rich foods with acidogenic foods are all good examples . Suggest that eating and drinking be followed by cariostatic foods such as xylitol chewing gum . * Drink water to satisfy thirst and hydration needs as often as possible . Restrict consumption of sweetened beverages to meal and snack times when they can be combined with other cariostatic foods . * When a patient reports excessive dietary intake of a fermentable carbohydrate to the point of displacing other important foods in the diet, identify alternatives that will help the patient maintain or achieve a healthy body weight, oral health status, and a nutrient-dense intake.

J Ind Microbiol Biotechnol, 2003 May, 30(5), 279 - 91 Epub 2003 Apr 16.
Hemicellulose bioconversion; Saha BC; Various agricultural residues, such as corn fiber, corn stover, wheat straw, rice straw, and sugarcane bagasse, contain about 20-40% hemicellulose, the second most abundant polysaccharide in nature . The conversion of hemicellulose to fuels and chemicals is problematic . In this paper, various pretreatment options as well as enzymatic saccharification of lignocellulosic biomass to fermentable sugars is reviewed . Our research dealing with the pretreatment and enzymatic saccharification of corn fiber and development of novel and improved enzymes such as endo-xylanase, beta-xylosidase, and alpha- l-arabinofuranosidase for hemicellulose bioconversion is described . The barriers, progress, and prospects of developing an environmentally benign bioprocess for large-scale conversion of hemicellulose to fuel ethanol, xylitol, 2,3-butanediol, and other value-added fermentation products are highlighted.

Appl Microbiol Biotechnol, 2003 May, 61(3), 257 - 60 Epub 2003 Mar 05.
Phosphate feeding strategy during production phase improves poly(3-hydroxybutyrate-co-3-hydroxyvalerate) storage by Ralstonia eutropha; Squio CR et al.; The effect of a phosphate feeding strategy and the optimal rate of biomass production ( r(x)) during the production phase of P(3HB-co-3HV) in a 6-l fermentor were determined in cultures of Ralstonia eutropha with the goal of enhancing polymer productivity . Rates of biomass production ( r(x)) between 0.00 and 0.20 gx r l(-1) h(-1) were monitored during the production phase . When a low rate of cell growth was maintained ( r(x) of 0.02 gx r l(-1) h(-1)), polymer production improved, resulting in a final cell mass, P(3HB-co-3HV) mass, and P(3HB-co-3HV) content of 98.2 g, 62.0 g and 63.1 wt%, respectively, after 27.3 h . The maximum polymer productivity obtained during the production phase was 1.36 g l(-1 )h(-1).

Appl Microbiol Biotechnol, 2003 May, 61(3), 197 - 205 Epub 2003 Jan 25.
Yeast flocculation: what brewers should know; Verstrepen KJ et al.; For many industrial applications in which the yeast Saccharomyces cerevisiae is used, e.g . beer, wine and alcohol production, appropriate flocculation behaviour is certainly one of the most important characteristics of a good production strain . Yeast flocculation is a very complex process that depends on the expression of specific flocculation genes such as FLO1, FLO5, FLO8 and FLO11 . The transcriptional activity of the flocculation genes is influenced by the nutritional status of the yeast cells as well as other stress factors . Flocculation is also controlled by factors that affect cell wall composition or morphology . This implies that, during industrial fermentation processes, flocculation is affected by numerous parameters such as nutrient conditions, dissolved oxygen, pH, fermentation temperature, and yeast handling and storage conditions . Theoretically, rational use of these parameters offers the possibility of gaining control over the flocculation process . However, flocculation is a very strain-specific phenomenon, making it difficult to predict specific responses . In addition, certain genes involved in flocculation are extremely variable, causing frequent changes in the flocculation profile of some strains . Therefore, both a profound knowledge of flocculation theory as well as close monitoring and characterisation of the production strain are essential in order to gain maximal control over flocculation . In this review, the various parameters that influence flocculation in real-scale brewing are critically discussed . However, many of the conclusions will also be useful in various other industrial processes where control over yeast flocculation is desirable.

Appl Microbiol Biotechnol, 2003 May, 61(3), 189 - 96 Epub 2003 Jan 14.
Aspergillus niger citric acid accumulation: do we understand this well working black box?
Karaffa L, Kubicek CP.
This Mini-Review summarizes the current knowledge on the biochemical and physiological events leading to massive citric acid accumulation by Aspergillus niger under industrially comparable conditions, thereby particularly emphasizing the roles of glycolytic flux and its control, excretion of citric acid from the mitochondria and the cytosol, and the critical fermentation variables . The potential of novel techniques for metabolic analysis and genomic approaches in understanding this fermentation is also discussed.

J Biotechnol, 2003 Apr 24, 102(2), 135 - 41
Enhanced conversion of soluble starch to trehalose by a mutant of Saccharomycopsis fibuligera sdu; Chi Z et al.; In our previous studies, it was found that Saccharomycopsis fibuligera sdu cells could accumulate 18.0% (gg(-1)) trehalose from soluble starch in SSY medium . However, the yeast strain contained high activities of acid and neutral trehalases, which were reported to mobilize trehalose accumulated by the cells during fermentation . In order to enhance the yield of trehalose, it is necessary to remove the trehalase activities from the cells . By mutagenesis of ethylmethanesulfonate, one mutant that assimilated trehalose very slowly, but grew on other carbon sources as fast as its parent strain, was isolated . In Biostat B2 2-1 fermentation, trehalose accumulation of the mutant was much higher than that of the wild type when grown in YPS medium containing starch . The activities of acid and neutral trehalases of this mutant were much lower than those of the wild type, respectively . We think the reduction of acid and neutral trehalase activities is considered to be responsible for the increased yield of trehalose accumulated by the mutant.

Chemosphere, 2003 Jun, 51(9), 893 - 900
Chemical oxidation of wastewater from molasses fermentation with ozone; Pena M et al.; Color removal from biologically pre-treated molasses wastewater by means of chemical oxidation with ozone has been investigated . Batch experiments have been performed in order to analyze the influence of ozone dosage and reaction time on color removal, molecular weight distribution and decolorization kinetics . Depending on the applied ozone dosage, color removal from 71% to 93% and COD reduction from 15% to 25% were reached after 30 min reaction time . TOC values remained constant throughout ozonation . Gel permeation chromatography corroborated that high molecular weight compounds, responsible for the brown color, were present in raw wastewater . UV spectral studies confirmed that these colored compounds were melanoidins . As a result of ozonation the concentration of chromophore groups decreased . Ozonation of synthetic melanoidin under the same experimental conditions provided similar color removal efficiencies . Pseudo-first order kinetics with respect to colored compounds were found.

Chemosphere, 2003 Jun, 51(9), 887 - 91
Detoxification of semisolid olive-mill wastes and pine-chip mixtures using Phanerochaete flavido-alba; Linares A et al.; Semisolid olive-mill residues, pine chips, and mixtures of both residues contain phytotoxic components capable of inhibiting germination and vegetative growth in plants . Solid-state cultures of Phanerochate flavido-alba on pine chips or mixtures of both residues reduce these phytotoxic effects in fermented substrates . The phenol and lipid contents in cultures detoxified by this fungus also decreases.

J Agric Food Chem, 2003 Apr 23, 51(9), 2727 - 32
Immunodetection of proteins from grapes and yeast in a white wine; Dambrouck T et al.; The objective of this study was to analyze the origin of proteins of a Chardonnay wine . Three various polyclonal antibodies raised against must, yeast, and bacteria proteins were produced . For microorganisms, only the secreted macromolecules were used . To this end, yeast and bacteria were cultured in a model medium under conditions close to those of winemaking . Results obtained using these specific antibodies indicate that most of the wine proteins came from grapes and many of them were glycoproteins . Some proteins of this Chardonnay wine came from the yeast; they were released during the alcoholic fermentation and consisted of high molecular weight mannoproteins . In contrast, no bacteria proteins were detected in this Chardonnay wine.

J Agric Food Chem, 2003 Apr 23, 51(9), 2625 - 31
Stable carbon isotopic composition of the wine and CO2 bubbles of sparkling wines: detecting C4 sugar additions; Martinelli LA et al.; Sparkling wines have become a popular beverage in recent years, and the production of these wines is subject to adulteration during fermentation . This study investigated the stable carbon isotopic composition (expressed as delta(13)C) of the wine and of the CO(2) bubbles produced during the second fermentation for a number of sparkling wines produced in different countries around the world . Carbon isotope ratio analyses were used to estimate the addition of sugar obtained from C(4) plants (sugar cane or corn) . The average delta(13)C values of the Brazilian brut, demi-sec, and doux sparkling wines were -20.5 +/- 1.2 per thousand (n = 18), -18.1 +/- 1.3 per thousand (n = 9), and -15.8 per thousand (n = 1), respectively . These values were statistically heavier (more positive carbon isotope ratio values) than the average delta(13)C of sparkling wines produced in other parts of South America (Argentina and Chile, -26.1 +/- 1.6 per thousand, n = 5) and Europe (France, Germany, Italy, Portugal, and Spain, -25.5 +/- 1.2 per thousand, n = 12), but not statistically different from sparkling wines produced in the United States or Australia . The most likely explanation for differences in the carbon isotope ratios of wines from these different regions is the addition of C(4) sugar during the production of some sparkling wines from Australia, Brazil, and the United States . The isotopic composition of the CO(2) bubbles (delta(13)C-CO(2)) followed similar trends . The average delta(13)C-CO(2) of most of the Brazilian and Argentine sparkling wines was -10.8 +/- 1.2 per thousand (n = 23), indicating that the likely source of carbon for the second fermentation was sugar cane . Conversely, the average delta(13)C-CO(2) of most of the sparkling wines produced in Chile and Europe was -22.0 +/- 1.2 per thousand (n = 13), suggesting that a different sugar (most likely sugar beet) was most used in the second fermentation . It was concluded that in many cases, the carbon isotope ratios of sparkling wine and CO(2) bubbles can provide valuable information about the sugar sources.

J Agric Food Chem, 2003 Apr 23, 51(9), 2539 - 43
Caffeoylsophorose in a red vinegar produced through fermentation with purple sweetpotato; Terahara N et al.; Recently, a new red vinegar has been developed via fermentation with the storage root of purple-fleshed sweetpotato, Ipomoea batatas L . cv . Ayamurasaki . The red vinegar had a higher antioxidative activity than white or black vinegars . The red vinegar contained some new components possibly derived from the original purple sweetpotato . A major component was isolated using preparative HPLC, and the chemical structure was determined to be 6-O-(E)-caffeoyl-(2-O-beta-d-glucopyranosyl)-alpha-d-glucopyranose (caffeoylsophorose) by MS and NMR . Because the caffeoylsophorose showed a high antioxidative activity, it plays an important functional role in red vinegar as do anthocyanins and other components . Examination of the mechanism of formation is now in progress.

Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet, 2002, 67(3), 611 - 6
The effects of spinosad, a naturally derived insect control agent, to the honeybee (Apis melifera); Miles M et al.; Spinosad is a novel insect control agent derived by fermentation of the Actinomycete bacterium, Saccharopolyspora spinosa . Spinosad controls many caterpillar pests in vines, pome fruit and vegetables (including tomatoes and peppers), thrips in tomatoes, peppers and ornamental cultivation and dipterous leafminers in vegetables and ornamentals . Application rates vary between 25 to 100 g of active substance per hectare (g as/ha) and 4.8 to 36 g of active substance per hectolitre (g as/hL) depending on the crop and target pest . It is important that plant protection products are authorized for use only in ways that do not pose an unacceptable risk of harm to honeybees . For this purpose testing was performed to enable the safety of spinosad to be evaluated . The effects of spinosad to honeybees have been extensively researched . Testing has been performed under a variety of conditions in a range of countries globally . Studies to determine the acute toxicity of spinosad under laboratory conditions were conducted to generate LD50 or LC50 values for oral and contact routes of administration . These demonstrated that spinosad was highly toxic to worker honeybees under worst case laboratory conditions and that the oral route of exposure provided the greater risk . Residue tests conducted under laboratory, semi-field and field conditions on worker honeybees foraging on treated foliage indicated that dry product residues were harmless . Therefore the effects seen in the laboratory acute toxicity tests did not translate to a more realistic exposure scenario indicating that safe use patterns for the product can be developed . Semi-field cage studies have also demonstrated that spinosad was safe to bees when applied to flowering crops during periods of bee activity . The majority of studies conducted have indicated that spinosad does not adversely affect honeybee behaviour, brood or queen . It can be concluded that spinosad when used according to the approved product label recommendations, would be safe to foraging worker bees, queen and brood . Additional levels of safety could be achieved by avoiding situations where bees would forage primarily on aphid honey dew.

Vet Microbiol, 2003 May 29, 93(3), 179 - 83
Shiga toxin-producing Escherichia coli (STEC) isolated from healthy dairy cattle in southern Brazil; Moreira CN et al.; Over a period of 1 year, the production of verotoxin was investigated in 1127 Escherichia coli isolated from 243 dairy cattle from 60 small farms in southern Brazil . Vero cell assay was used to detect toxins in culture supernatants from E . coli isolated from bovine feces . Shiga toxin-producing E . coli (STEC) detection rates were 95% (57 of 60) for farms and 49% (119 of 243) for cattle . Prevalence of STEC-positive cattle in the farms ranged from 0 to 100% . Ninety-six percent (315 of 327) of the STEC isolates did not react in the panel of sera used for typing . Twelve isolates, all non-motile, belonged to serogroups previously associated with human diseases, and 67% (8 of 12) were of only two serotypes (O91:H- and sorbitol-fermenting O157:H-) . These results indicate that dairy cattle from the region surveyed may be a source of STEC potentially pathogenic for humans.

Int J Pharm, 2003 Apr 30, 256(1-2), 199 - 206
Process for producing the potential food ingredient DFA III from inulin: screening, genetic engineering, fermentation and immobilisation of inulase II; Jahnz U et al.; Difructose anhydride (DFA III) is a new potential sweet food additive . A screening was undertaken to isolate bacterial strains for conversion of inulin to DFA . Of special interest were thermotolerant enzymes . Some 400 strains were investigated, among four of them produce DFA and strain Buo141 expresses an extracellular enzyme which is stable at elevated temperatures . Based on metabolic data and 16S-rRNA-sequencing, the strain was identified as a new Arthrobacter species . For increased enzyme production, the inulase gene was cloned into E . coli XL1-blue, inulase II was expressed and its activity detected . After identifying the cleavage site, the sequence coding for a signal-peptide was eliminated from the plasmid and a beneficial amino acid exchange introduced by error-prone PCR . The recombinant E . coli was fermented to 10.5 g/l and after disruption an activity of 1.76 MioU/l was observed . The enzyme was flocculated from supernatant and entrapped in calcium alginate hydrogels . To enable production of uniform and small beads JetCutter technology was used with a production rate of 5600 beads/(snozzle) . The influence of bead diameter on activity was investigated . An activity of 196 U/g was measured for 600-microm beads.

Eur J Biochem, 2003 Apr, 270(8), 1863 - 74
Complex II from phototrophic purple bacterium Rhodoferax fermentans displays rhodoquinol-fumarate reductase activity; Miyadera H et al.; It has long been accepted that bacterial quinol-fumarate reductase (QFR) generally uses a low-redox-potential naphthoquinone, menaquinone (MK), as the electron donor, whereas mitochondrial QFR from facultative and anaerobic eukaryotes uses a low-redox-potential benzoquinone, rhodoquinone (RQ), as the substrate . In the present study, we purified novel complex II from the RQ-containing phototrophic purple bacterium, Rhodoferax fermentans that exhibited high rhodoquinol-fumarate reductase activity in addition to succinate-ubiquinone reductase activity . SDS/PAGE indicated that the purified R . fermentans complex II comprises four subunits of 64.0, 28.6, 18.7 and 17.5 kDa and contains 1.3 nmol heme per mg protein . Phylogenetic analysis and comparison of the deduced amino acid sequences of R . fermentans complex II with pro/eukaryotic complex II indicate that the structure and the evolutional origins of R . fermentans complex II are closer to bacterial SQR than to mitochondrial rhodoquinol-fumarate reductase . The results strongly indicate that R . fermentans complex II and mitochondrial QFR might have evolved independently, although they both utilize RQ for fumarate reduction.

Appl Microbiol Biotechnol, 2003 Jun, 61(5-6), 435 - 40 Epub 2003 Apr 12.
Biotechnological solubilization of rock phosphate on media containing agro-industrial wastes; Vassilev N et al.; Rock phosphate (RP) is an important natural material traditionally used for the production of phosphorus (P) fertilizers . Compared with chemical treatment, microbial solubilization of RP is an alternative environmentally mild approach . An overview of biotechnological techniques, mainly based on solubilization processes involving agro-industrial residues, is presented . Potential advantages of composting, solid-state fermentation, and liquid submerged fermentation employing free and immobilized microorganisms that produce organic acids and simultaneously solubilize RP are discussed . Subsequent introduction of the final fermented products into soil-plant systems promotes plant growth and P acquisition.

J Dermatol, 2003 Feb, 30(2), 91 - 7
Role of foods in irregular aggravation of atopic dermatitis; Uenishi T et al.; Although it is well known that patients with atopic dermatitis often show unpredictable, irregular aggravation of skin lesions, there are no previously published studies examining trigger factors for such unpredictable aggravation . We investigated whether foods play a role in the unpredictable, irregular worsening of atopic dermatitis . The patient group included 195 Japanese adult patients with atopic dermatitis who showed unpredictable, irregular aggravation of skin lesions . They were hospitalized and openly challenged with suspected foods . Photographs of representative skin lesion sites were taken at baseline and before and after the challenge . Challenge-positive foods were determined by evaluating the comparable before-after challenge photographs . One to three (average: 1.7) challenge-positive foods were confirmed in 86 (44%) of the 195 patient examined . Predominant offending foods were chocolate, cheese, coffee, yogurt and some Japanese foods such as glutinous rice cake, soy sauce and fermented soybeans . Specific IgE values to the offending foods were mostly negative . Patients were asked to exclude challenge-positive foods from their diets . They were then discharged and followed up for 3 months at our outpatient clinic . Exclusion of the offending foods for 3 months brought about a progressive improvement of the disease . These results suggest that foods play an important role in unpredictable, irregular aggravation of skin lesions in patients with atopic dermatitis.

Appl Microbiol Biotechnol, 2004 Jan, 63(4), 431 - 7 Epub 2003 Apr 11.
Characterization of Aspergillus oryzae fermentation extract effects on the rumen fungus Neocallimastix frontalis, EB 188 . Part 2 . Carbon source utilization and effects on zoospore production; Schmidt JA et al.; The effect of a commercial Aspergillus oryzae fermentation extract on the utilization of carbon source and zoospore production by the rumen fungus Neocallimastix frontalis EB 188 was determined . In addition, the composition of a soluble extract prepared from the commercial product was analyzed . This extract was added to N . frontalis EB 188 cultures grown on a variety of substrates and periodically assayed for protein, enzymes, zoospore production, and carbon source utilization . The powdered product contained 93% dry matter, more than 3,000 A . oryzaespores per gram, and did not contain strong buffers or high concentrations of salt . Measurable concentrations of DNA, protein, carbohydrate and several enzymes including cellulase and amylase were also found . Soluble extract increased fungal physiology and treated cultures produced significantly higher levels of supernatant protein and enzymes including amylase, cellulase and beta-glucosidase . The fungal response depended on culture carbon source . However, culture zoospore production was increased regardless of substrate provided . Culture utilization of glucose was more rapid in treated cultures, yet high levels of the extract greatly inhibited glucose utilization.

Appl Microbiol Biotechnol, 2004 Jan, 63(4), 422 - 30 Epub 2003 Apr 11.
Characterization of Aspergillus oryzae fermentation extract effects on the rumen fungus Neocallimastix frontalis, EB 188 . Part 1 . Zoospore development and physiology; Schmidt JA et al.; Experiments were performed to determine the effect of Aspergillus oryzae (AO) fermentation extract on zoospore development in the rumen fungus Neocallimastix frontalis EB 188 . Powdered product, or liquid extract prepared from such powder, was added at the recommended value for supplementation in dairy cattle . Stationary and stirred cultures were periodically sampled and assayed for extracellular and intracellular protein and enzymes, gas production, zoospore production and maturation, and carbon source utilization . Soluble extract increased fungal physiology when grown in stirred vessels or stationary cultures . Treated cultures produced higher levels of enzymes (nearly double) . Mobile zoospores matured into germination entities more rapidly in treated cultures, and when powdered product was used, nearly 3 times more motile zoospores were produced at 56 h of fungal growth . Levels of the intracellular enzyme malate dehydrogenase increased by 6-fold in the presence of powdered product . Product wheat bran carrier used as soluble extract or powder had very little effect on fungal cultures . Medium cellulose was completely hydrolyzed in all cultures but this occurred earlier in those containing AO treatment.

FEMS Yeast Res, 2003 May, 3(3), 269 - 87
Transcriptional profiling of wine yeast in fermenting grape juice: regulatory effect of diammonium phosphate; Marks VD et al.; The nitrogen composition of grape musts affects fermentation kinetics and production of aroma and spoilage compounds in wine . It is common practice in wineries to supplement grape musts with diammonium phosphate (DAP) to prevent nitrogen-related fermentation problems . Laboratory strains of Saccharomyces cerevisiae preferentially use rich nitrogen sources, such as ammonia, over poor nitrogen sources . We used global gene expression analysis to monitor the effect of DAP addition on gene expression patterns in wine yeast in fermenting Riesling grape must . The expression of 350 genes in the commercial wine yeast strain VIN13 was affected; 185 genes were down-regulated and 165 genes were up-regulated in response to DAP . Genes that were down-regulated encode small molecule transporters and nitrogen catabolic enzymes, including those linked to the production of urea, a precursor of ethyl carbamate in wine . Genes involved in amino acid metabolism, assimilation of sulfate, de novo purine biosynthesis, tetrahydrofolate one-carbon metabolism, and protein synthesis were up-regulated . The expression level of 86 orphan genes was also affected by DAP.

FEMS Yeast Res, 2003 May, 3(3), 239 - 45
Cyanide-resistant respiration, a very frequent metabolic pathway in yeasts; Veiga A et al.; It has recently been shown that cyanide-resistant respiration (CRR) is very common in Crabtree-negative yeasts (incapable of aerobic fermentation) and in non-fermentative yeasts . It is conferred by a salicylhydroxamic acid-sensitive alternative oxidase that transfers electrons from ubiquinol to oxygen, bypassing the cytochrome chain . An interesting finding is that, in general, whenever CRR is present, complex I is also present . In this article we briefly review the occurrence of CRR, the biochemistry and molecular biology of the alternative oxidase, and summarise the putative functions that have been attributed to this ubiquitous metabolic pathway, whose usefulness for the yeast cells still remains obscure.

Biodegradation, 2002, 13(5), 329 - 42
Reduction of Np(V) and precipitation of Np(IV) by an anaerobic microbial consortium; Rittmann BE et al.; A combination of experimental, analytical, and modeling investigations shows that an anaerobic, sulfate-reducing consortium reduced Np(V) to Np(IV), with subsequent precipitation of a Np(IV) solid . Precipitation of Np(IV) during growth on pyruvate occurred before sulfate reduction began . H2 stimulated precipitation of Np(IV) when added alone to growing cells, but it slowed precipitation when added along with pyruvate . Increasing concentrations of pyruvate also retarded precipitation . Accumulation of an intermediate pyruvate-fermentation product--probably succinate--played a key role in retarding Np(IV) precipitation by complexing the Np(IV) . Hydrogen appears to have two roles in controlling Np precipitation: donating electrons for Np(V) reduction and modulating intermediate levels . That Np(V) is microbially reduced and subsequently precipitated under anaerobic conditions has likely beneficial implications for the containment of Np on lands contaminated by radionuclides, but complexation by fermentation intermediates can prevent immobilization by precipitation.

Bioresour Technol, 2003 Feb, 86(3), 227 - 33
Stimulatory effect of alcohols (methanol and ethanol) on citric acid productivity by a 2-deoxy D-glucose resistant culture of aspergillus niger GCB-47; Haq IU et al.; The present study describes citric acid fermentation by Aspergillus niger GCB-47 in a 15-1 stainless steel stirred fermentor . Among the alcohols tested as stimulating agents, 1.0% (v/v) methanol was found to give maximum amount of anhydrous citric acid (90.02 +/- 2.2 g/l), 24 h after inoculation . This yield of citric acid was 1.96 fold higher than the control . Methanol has a direct effect on mycelial morphology and it promotes pellet formation . It also increases the cell membrane permeability to provoke more citric acid excretion from the mycelial cells . The sugar consumed and % citric acid was 108 +/- 3.8 g/l and 80.39 +/- 4.5%, respectively . The desirable mycelial morphology was in the form of small round pellets having dry cell mass 14.5 +/- 0.8 g/l . Addition of ethanol, however, did not found to enhance citric acid production, significantly . The maximum value of Yp/x (i.e., 5.825 +/- 0.25 g/g) was observed when methanol was used as a stimulating agent . The best results of anhydrous citric acid were observed, 6 days after inoculation when the initial pH of fermentation medium was kept at 6.0.

J Gen Appl Microbiol, 2002 Dec, 48(6), 329 - 34
Improvement of industry-applied rifamycin B-producing strain, Amycolatopsis mediterranei, by rational screening; Jin ZH et al.; An industrially applied rifamycin B-producing strain, Amycolatopsis mediterranei XC 1-02, was used for further screening . A special mutation and screening procedure was adopted to select a strain, which can alleviate the inhibition caused by both aromatic amino acid and p-hydroxybenzoic acid in the pathway of rifamycin B biosynthesis as well as enhance the production of propionate, one of the precursors of rifamycin B biosynthesis . By the above methods, a strain A . mediterranei XC 9-25 was obtained, and its rifamycin B productivity in shaking flask reaches 10 g/L, which is 2.38 times higher than that of the ancestral strain XC 1-02 . The productivity of rifamycin B fed-batch fermentation in 60000 L fermentor with A . mediterranei XC 9-25 reached 19.11 g/L.

Lett Appl Microbiol, 2003, 36(5), 302 - 6
Effect of volume of culture medium on enhanced citric acid productivity by a mutant culture of Aspergillus niger in stirred fermentor; Haq IU et al.; AIMS: The present study deals with the effect of volume of culture medium on enhanced citric acid productivity by a mutant culture of Aspergillus niger . METHODS AND RESULTS: A laboratory scale stirred fermentor of 15-l capacity was employed for all microbial cultivations . Blackstrap molasses, a by-product of sugar industries is easily and abundantly available for its exploitation as a carbon source in fermentation processes . The parental culture of A . niger was improved by mutation using ultraviolet radiations and N-methyl N-nitro N-nitroso guanidine, i.e . mutagen MNNG . Six MUV and eight MNNG-treated mutant strains were isolated after extensive screening and optimization . Mutant strain of A . niger MNNG-2 showed enhanced citric productivity (87.60 g l-1) over the parental strain BTL-45 (19.53 g l-1) and other mutant derivatives (49.85 g l-1 citric acid in case of mutant MUV-5 and 76.82 g l-1 in case of mutant MNNG-7) . The optimal sugar level was found to be 150 g l-1 (optimum volume of the medium, 60%) after 6 days of inoculation, which is economically significant . Specific productivity of the mutant culture MNNG-2 (qp = 0.057 g/g cells h-1) was several folds higher than other strains . SIGNIFICANCE AND IMPACT OF THE STUDY: The results of the present study are of commercial level . All kinetic parameters including yield coefficients and volumetric rates revealed the hyper-producibility of citric acid by mutant MNNG-2 using blackstrap molasses as the basal medium in stirred fermentor.

Lett Appl Microbiol, 2003, 36(5), 268 - 71
The importance of amino acids as carbon sources for Micromonospora echinospora (ATCC 15837); Hoskisson PA et al.; AIMS: This study set out to investigate the effect of amino acids on the uptake of glucose by Micromonospora eichinospora (ATCC 15837) . METHODS AND RESULTS: The specific rate of glucose uptake was found to be reduced when organic nitrogen components were present in the medium . Radioactive uptake studies revealed that the Km for glucose in this organism was 53 mm, indicating a low affinity for uptake compared with other actinomycete sugar transport systems . Individual amino acids negatively influenced the rate of glucose transport, suggesting a relationship between amino acid metabolism and glucose uptake in this organism . The sugar transport system was found to be an active process being inhibited by ionophores and KCN . CONCLUSIONS: The data suggest a direct link between amino acid metabolism and glucose uptake at the level of sugar transport . SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that the uptake of glucose, a major carbon source for many antibiotic fermentations, is significantly reduced in the presence of amino acids . This fact should inform the medium design and feeding regimes of fermentations involving similar actinomycetes.

J Biotechnol, 1999 Jun 11, 72(1-2), 213 - 28
Derepression of galactose metabolism in melibiase producing bakers' and distillers' yeast; Ronnow B et al.; Beet molasses is widely used as a growth substrate for bakers' and distillers' yeast in the production of biomass and ethanol . Most commercial yeasts do not fully utilise the carbohydrates in molasses since they are incapable of hydrolysing the disaccharide melibiose to glucose and galactose . Also, expression of genes encoding enzymes for the utilisation of carbon sources that are alternatives to glucose is tightly regulated, sometimes rates of yeast growth and/or ethanol production . The GAL genes are regulated by specific induction by galactose and repression during growth on glucose . In an industrial distillers' yeast, two genes interacting synergistically in glucose repression of galactose utilization, MIG1 and GAL80, have been disrupted with MEL1, encoding melibiase . The physiology of the wild-type strain and the recombinant strains was investigated on mixtures of glucose and galactose and on molasses . The recombinant strain started to ferment galactose when 9.7 g 1(-1) glucose was still present during a batch fermentation, whereas the wild-type strain did not consume any galactose in the presence of glucose . The ethanol yield in the recombinant strain was 0.50 g ethanol g sugar (-1) in an ethanol fermentation on molasses, compared with 0.48 g ethanol g sugar (-1) for the wild-type strain . The increased ethanol yield was due to utilization of melibiose in the molasses.

J Ind Microbiol Biotechnol, 2003 Apr, 30(4), 220 - 4 Epub 2003 Apr 08.
Isolation and insecticidal activity of mellamide from Aspergillus melleus; Ondeyka JG et al.; Mellamide, a novel indole amide, was isolated from a fermentation of Aspergillus melleus using silica gel and high-performance liquid chromatographic methods . This allowed its separation from three known antiparasitic compounds (ochratoxin A, viomellin and xanthomegnin) also present in the potent extract . The structure was elucidated by (1)H, (13)C, COSY, DEPT, HMQC and HMBC NMR experiments . HR-FTMS aided in the molecular weight and formula determination . Mellamide showed in vitro insecticidal activity in bioassays against larvae of Lucilia sericata and Aedes egypti with LD(90) of 1,000 and 50 micro g/ml, respectively.

J Ind Microbiol Biotechnol, 2003 Aug, 30(8), 472 - 9 Epub 2003 Apr 05.
Dereplication of Streptomyces soil isolates and detection of specific biosynthetic genes using an automated ribotyping instrument; Ritacco FV et al.; The discrimination of distinct cultures among morphologically similar Streptomyces soil isolates (dereplication) and the detection of specific biosynthetic pathways in these strains are important steps in the selection of microorganisms to include in a natural products library . We have developed methods for analysis of actinomycetes using the RiboPrinter microbial characterization system, an automated instrument that performs ribotyping on bacterial samples . To evaluate our dereplication method, 26 Streptomyces isolates, obtained from soil samples collected in Maui, Hawaii, were ribotyped and compared with each other, using the RiboPrinter . The strains were also compared by 16S rDNA sequence analysis, MIDI fatty acid analysis, and LC-MS profiling of fermentation extracts . The RiboPrinter was able to identify closely related isolates and to discriminate between morphologically similar isolates with unique genetic, fatty acid and fermentation profiles . For the detection of biosynthetic genes, a 1,006-bp probe containing a portion of an adenylation domain of a non-ribosomal peptide synthetase (NRPS) was employed . Using this alternate probe in place of the standard ribosomal probe, the RiboPrinter was able to detect NRPS genes in several strains of Streptomyces . These results demonstrate that the RiboPrinter has multiple applications in a natural products research program.

J Ind Microbiol Biotechnol, 2003 Mar, 30(3), 150 - 6 Epub 2003 Mar 25.
Production of the glycopeptide antibiotic A40926 by Nonomuraea sp . ATCC 39727: influence of medium composition in batch fermentation; Gunnarsson N et al.; Nonomuraea sp . ATCC 39727 is a novel actinomycete species and the producer of A40926, a glycopeptide antibiotic structurally similar to teichoplanin . In the present study, a defined minimal medium was designed for Nonomuraea fermentation . The influence of initial phosphate, glucose and ammonium concentrations on antibiotic productivity was investigated in batch fermentation and the effect of glucose limitation was studied in fed-batch fermentation . It was found that low initial concentrations of phosphate and ammonium are beneficial for A40926 production and that productivity is not enhanced during glucose limitation . Furthermore, the initiation of A40926 production was not governed by residual ammonium and phosphate concentrations, although the level of these nutrients strongly influenced A40926 production rates and final titers.

Eukaryot Cell, 2003 Apr, 2(2), 341 - 50
Histone H1 Is required for proper regulation of pyruvate decarboxylase gene expression in Neurospora crassa; Folco HD et al.; We show that Neurospora crassa has a single histone H1 gene, hH1, which encodes a typical linker histone with highly basic N- and C-terminal tails and a central globular domain . A green fluorescent protein-tagged histone H1 chimeric protein was localized exclusively to nuclei . Mutation of hH1 by repeat-induced point mutation (RIP) did not result in detectable defects in morphology, DNA methylation, mutagen sensitivity, DNA repair, fertility, RIP, chromosome pairing, or chromosome segregation . Nevertheless, hH1 mutants had mycelial elongation rates that were lower than normal on all tested carbon sources . This slow linear growth phenotype, however, was less evident on medium containing ethanol . The pyruvate decarboxylase gene, cfp, was abnormally derepressed in hH1 mutants on ethanol-containing medium . This derepression was also found when an ectopically integrated fusion of the cfp gene promoter to the reporter gene hph was analyzed . Thus, Neurospora histone H1 is required for the proper regulation of cfp, a gene with a key role in the respiratory-fermentative pathway.

Biotechnol Appl Biochem, 2003 Jun, 37(Pt 3), 207 - 18
Scaleable processes for the manufacture of therapeutic quantities of plasmid DNA; Shamlou PA; The need for scaleable processes to manufacture therapeutic plasmid DNA (pDNA) is easy to overlook when attention is focused primarily on vector design and establishment of early clinical results . pDNA is a large molecule and has properties that are similar to those of the contaminating chromosomal DNA . These, combined with the low initial concentration of plasmids in the host cell, provide unique process challenges that require significant upfront design to establish robust manufacturing processes that can also comply with current Good Manufacturing Practice ('cGMP') and produce milligram-to-kilogram quantities of pDNA product . This review describes promising scaleable processes that are currently being assessed for production of therapeutic supercoiled pDNA . Fermentation strategies for improving supercoiled plasmid yield and reducing contaminant concentrations are reviewed, and downstream processes are assessed for their ability to efficiently remove cellular contaminants, separate the supercoiled form of the pDNA from its open circular and linear forms, and prepare the purified drug substance for formulation . Current strategies are presented for developing stable delivery systems, and approaches to quality assurance and quality control are discussed.

Appl Microbiol Biotechnol, 2003 Oct, 62(5-6), 564 - 8 Epub 2003 Apr 03.
Characterization of the metabolic shift of Saccharomyces bayanus var . uvarum by continuous aerobic culture; Serra A et al.; Saccharomyces bayanus, being of interest for wine-making, is not as well known as S . cerevisiae and, due to many changes in the yeast classification, accurate data concerning its metabolic activity are difficult to find . In order to produce this yeast as an active dry yeast to be used as a starter in wine-making, its sensitivity to glucose was determined as the objective of our work . Using the pulse technique in continuous culture, it was found that growth in a synthetic medium was not limited by vitamins or mineral salts . We determined the critical dilution rate of a continuous culture and performed an aerobic continuous culture, measuring the respiratory quotient on-line in order to observe the metabolic shift from respiratory to fermentative metabolism . The S . bayanus var . uvarum strain studied was Crabtree-positive (glucose-sensitive) but had a weaker respiratory capacity than S . cerevisiae since the dilution rate of the metabolic shift was only 0.15 h(-1) . These new data provide essential information for the biomass production of this yeast strain for wine-making.

Appl Microbiol Biotechnol, 2003 Aug, 62(2-3), 156 - 62 Epub 2003 Apr 03.
Efficient cellulase production by Trichoderma reesei in continuous cultivation on lactose medium with a computer-controlled feeding strategy; Bailey MJ et al.; A low-foaming hydrophobin II deletant of the Trichoderma reesei strain Rut-C30 was used for production of cellulases by continuous cultivation on lactose medium in a laboratory fermenter . The control paradigm of the addition of new medium to the continuous process was based on the growth dynamics of the fungus . A decrease in the rate of base addition to the cultivation for pH-minimum control was used as an indicator of imminent exhaustion of carbon source for growth and enzyme induction . When the amount of base added per 5 min computation cycle decreased below a given value, new medium was added to the fermenter . When base addition for pH control thereafter increased above the criterion value, due to increased growth, the medium feed was discontinued or decreased . The medium feeding protocol employed was successful in locking the fungus in the stage of imminent, but not actual, exhaustion of carbon source . According to the results of a batch cultivation of the same strain on the same medium, this is the phase of maximal enzyme productivity . The medium addition protocol used in this work resulted in a very stable continuous process, in which cellulase productivity was maintained for several hundred hours at the maximum level observed in a batch cultivation for only about 10 h . Despite a major technical disturbance after about 420 h, the process was restored to stability . When the cultivation was terminated after 650 h, the level of enzyme production was still maximal, with no signs of instability of the process.

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