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 an