Mead Production by Saccharomyces cerevisiae Safbrew T-58 and Saccharomyces bayanus (Premier Blanc and Premier Cuvée): Effect of Cowpea (Vigna unguiculata L. Walp) Extract Concentration.

This work evaluated the effect of the cowpea (Vigna unguiculata L. Walp) extract concentration on mead production. Fermentations were carried out in 500-mL Erlenmeyer flasks containing 250 mL of honey wort (30 °Brix), supplemented with 1 g L-1 of ammonium sulfate and 0.1 g L-1 of magnesium chloride and the cowpea extract (5 and 30 g L-1), inoculated with 106 cells mL-1, and incubated at 30 °C for 240 h. Higher cell growth ((cells mL-1): 11.1 × 107, 11.3 × 107, and 19.6 × 107; substrate consumption (%): 86.0, 90.0, and 85.0) and ethanol production ((v v-1 %): 15.0, 15.5, and 14.1) for yeasts Safbrew T-58, Premier Blanc, and Premier Cuvée, respectively, were obtained with 30 g L-1 of bean extract. S. bayanus Premier Blanc had the best metabolic activity with lowest glycerol production (8.5 g L-1) and highest ethanol volumetric yields (0.51-1.52 h-1) after 48 h of fermentation.

Amino Acid Supplementations Enhance the Stress Resistance and Fermentation Performance of Lager Yeast During High Gravity Fermentation.

The effects of different wort gravity or ethanol concentration in initial wort on the fermentation performance of lager yeast and assimilation of free amino acids (FAAs) were studied. Results showed that compared with high wort gravity (24°P), high ethanol concentration (10%, v/v) decreased yeast growth, cell viability, and wort fermentability significantly. The assimilation of FAAs was changed dramatically by high ethanol toxicity, and positive correlations between the assimilation amounts of 10 FAAs (Asp, Ser, Gly, Arg, Tyr, Val, Met, Lys, Ile, and Leu) and fermentation performance (cell viability, fermentability, and ethanol production) were identified, especially for Arg and Lys exhibiting extremely significant positive correlations. Furthermore, confirmatory testing was carried out by supplementing 24°P worts with 10 FAAs of 0.5, 1, and 2 times of their standard concentrations, respectively. Results exhibited that 10 FAA supplementations improved physiological characteristics and fermentation performance of lager yeast significantly, especially for 1 times FAA supplementation increasing wort fermentability and ethanol yield by 6 and 17%, respectively, and upregulated the expression level of HSP12 and increased more intracellular trehalose accumulation in yeast cells, indicating that stronger protective function was stimulated in yeast cells. Therefore, it was suggested that these 10 FAAs could regulate yeast cells to adapt to high gravity environmental stresses.

Functional anatomy of the colonic bioreactor: Impact of antibiotics and Saccharomyces boulardii on bacterial composition in human fecal cylinders.

Sections of fecal cylinders were analyzed using fluorescence in situ hybridization targeting 180 bacterial groups. Samples were collected from three groups of women (N=20 each) treated for bacterial vaginosis with ciprofloxacin+metronidazole. Group A only received the combined antibiotic regimen, whereas the A/Sb group received concomitant Saccharomyces boulardii CNCM I-745 treatment, and the A_Sb group received S. boulardii prophylaxis following the 14-day antibiotic course. The number of stool cylinders analyzed was 188 out of 228 in group A, 170 out of 228 in group A/Sb, and 172 out of 216 in group A_Sb. The colonic biomass was organized into a separate mucus layer with no bacteria, a 10-30µm broad unstirred transitional layer enriched with bacteria, and a patchy fermentative area that mixed digestive leftovers with bacteria. The antibiotics suppressed bacteria mainly in the fermentative area, whereas abundant bacterial clades retreated to the transitional mucus and survived. As a result, the total concentration of bacteria decreased only by one order. These effects were lasting, since the overall recovery of the microbial mass, bacterial diversity and concentrations were still below pre-antibiotic values 4 months after the end of antibiotic treatment. Sb-prophylaxis markedly reduced antibiotic effects and improved the recovery rates. Since the colon is a sophisticated bioreactor, the study indicated that the spatial anatomy of its biomass was crucial for its function.

Effects of Lys and His supplementations on the regulation of nitrogen metabolism in lager yeast.

Significant positive correlations between wort fermentability and the assimilation of Lys and His under normal-gravity and high-gravity conditions indicated that Lys and His were the key amino acids for lager yeast during beer brewing. In order to obtain insight into the roles of Lys and His in nitrogen regulation, the influences of Lys, His and their mixture supplementations on the fermentation performance and nitrogen metabolism in lager yeast during high-gravity fermentation were further investigated in the present study. Results showed that Lys and His supplementations improved yeast growth, wort fermentability, ethanol yield and the formation of flavor volatiles. Lys supplementation up-regulated Ssy1p-Ptr3p-Ssy5p (SPS)-regulated genes (LYP1, HIP1, BAP2 and AGP1) dramatically compared to nitrogen catabolite repression (NCR)-sensitive genes (GAP1 and MEP2), whereas His supplementation activated SPS-regulated genes slightly in exponential phase, and repressed NCR-sensitive genes significantly throughout the fermentation. Lys and His supplementations increased the consumption of Glu and Phe, and decreased the consumption of Ser, Trp and Arg. Moreover, Lys and His supplementations exhibited similar effects on the fermentation performance, and were more effective than their mixture supplementation when the same dose was kept. These results demonstrate that both Lys and His are important amino acids for yeast nitrogen metabolism and fermentation performance.

Saccharomyces boulardii does not prevent relapse of Crohn's disease.

BACKGROUND & AIMS: Saccharomyces boulardii is a probiotic yeast that has been shown to have beneficial effects on the intestinal epithelial barrier and digestive immune system. There is preliminary evidence that S boulardii could be used to treat patients with Crohn's disease (CD). We performed a randomized, placebo-controlled trial to evaluate the effects of S boulardii in patients with CD who underwent remission during therapy with steroids or aminosalicylates. METHODS: We performed a prospective study of 165 patients who achieved remission after treatment with steroids or salicylates; they were randomly assigned to groups given S boulardii (1 g/day) or placebo for 52 weeks. The primary end point was the percentage of patients in remission at week 52. Time to relapse, Crohn's disease activity index scores, and changes in parameters of inflammation were secondary end points. RESULTS: CD relapsed in 80 patients, 38 in the S boulardii group (47.5%) and 42 in the placebo group (53.2%, a nonsignificant difference). The median time to relapse did not differ significantly between patients given S boulardii (40.7 weeks) vs placebo (39.0 weeks). There were no significant differences between groups in mean Crohn's disease activity index scores or erythrocyte sedimentation rates or in median levels of C-reactive protein. In a post hoc analysis, nonsmokers given S boulardii were less likely to experience a relapse of CD than nonsmokers given placebo, but this finding requires confirmation. CONCLUSIONS: Although the probiotic yeast S boulardii is safe and well tolerated, it does not appear to have any beneficial effects for patients with CD in remission after steroid or salicylate therapies.

The effect of probiotics and mucoprotective agents on PPI-based triple therapy for eradication of Helicobacter pylori.

BACKGROUND/AIMS: Recent studies have found that probiotics have anti-Helicobacter pylori (HP) properties. We evaluated the additive effects of (i) Saccharomyces boulardii combined with proton pump inhibitor (PPI)-based triple therapy and (ii) S. boulardii and a mucoprotective agent (DA-9601) coupled with PPI-based triple therapy for HP eradication. METHODS: We recruited 991 HP infected patients and randomized them into one of three groups, (A) PPI-based 7-day triple therapy, (B) the same triple therapy plus S. boulardii for 4 weeks, and (C) the same 7-day triple therapy plus S. boulardii and mucoprotective agent for 4 weeks. All patients in the three groups were tested via (13)C-urea breath test 4 weeks after the completion of the therapy. RESULTS: According to the results of an intention-to-treat analysis, HP eradication rates for the groups A, B, and C were 71.6% (237/331), 80.0% (264/330), and 82.1% (271/330), respectively (p = .003). According to the results of a per protocol analysis, the eradication rates were 80.0% (237/296), 85.4% (264/309) and, 84.9% (271/319), respectively (p = .144). The frequency of side effects in group B (48/330) and C (30/330) was lower than that in group A (63/331) (p < .05). CONCLUSION: This study suggests that supplementation with S. boulardii could be effective for improving HP eradication rates by reducing side effects thus helping completion of eradication therapy. However, there were no significant effects on HP eradication rates associated with the addition of mucoprotective agents to probiotics and triple therapy.

Growth and biosurfactant synthesis by Nigerian hydrocarbon-degrading estuarine bacteria

The ability of microorganisms to degrade petroleum hydrocarbons is important for finding an environmentally-friendly method to restoring contaminated environmental matrices. Screening of hydrocarbon-utilizing and biosurfactant-producing abilities of organisms from an estuarine ecosystem in Nigeria, Africa, resulted in the isolation of five microbial strains identified as Corynebacterium sp. DDv1, Flavobacterium sp. DDv2, Micrococcus roseus DDv3, Pseudomonas aeruginosa DDv4 and Saccharomyces cerevisae DDv5. These isolates grew readily on several hydrocarbons including hexadecane, dodecane, crude oil and petroleum fractions. Axenic cultures of the organisms utilized diesel oil (1.0 % v/v) with generation times that ranged significantly (t-test, P < 0.05) between 3.25 and 3.88 day, with concomitant production of biosurfactants. Kinetics of growth indicates that biosurfactant synthesis occurred predominantly during exponential growth phase, suggesting that the bioactive molecules are primary metabolites. Strains DDv1 and DDv4 were evidently the most metabolically active in terms of substrate utilization and biosurfactant synthesis compared to other strains with respective emulsification index of 63 and 78 %. Preliminary biochemical characterization indicates that the biosurfactants are heteropolymers consisting of lipid, protein and carbohydrate moieties. The hydrocarbon catabolic properties coupled with biosurfactant-producing capabilities is an asset that could be exploited for cleanup of oil-contaminated matrices and also in food and cosmetic industries. Rev. Biol. Trop. 56 (4): 16031611. Epub 2008 December 30.

La capacidad de los microorganismos para degradar hidrocarburos del petróleo es de gran importancia para hallar un método aceptable y ambientalmente amigable para la restauración de terrenos ambientalmente contaminados. Al investigar las capacidades de los organismos de un ecosistema de estuario que utilizan hidrocarburos y producen biosurfactantes, se produjo como resultado el aislamiento de cinco cepas microbianas identificadas como Corynebacterium sp. DDv1, Flavobacterium sp. DDv2, Micrococcus roseus DDv3, Pseudomonas aeruginosa y DDv4 Saccharomyces cerevisiae DDv5. Estas cepas crecieron fácilmente en varios hidrocarburos incluyendo hexadecanos, dodecanos, petróleo crudo y fracciones de petróleo. Los cultivos axénicos de organismos utilizaron diesel (1.0% v/v) con períodos por generación con ámbitos significativos (t-test, P <0.05) de entre 3.25 y 3.88 días, con la consiguiente producción de bio-surfactantes. La cinética del crecimiento indica que la síntesis de bio-surfactante se produjo principalmente durante la fase de crecimiento exponencial, lo que sugiere que las moléculas bioactivas son metabolitos primarios. Las cepas DDv1 y DDv4 fueron evidentemente las más metabólicamente activas en términos de utilización del sustrato y la síntesis de bio-surfactantes en comparación con otras cepas con índices respectivos de emulsificación de 63 y 78%. La caracterización bioquímica preliminar indica que los bio-surfactantes son heteropolímeros constituidos de fracciones de lípidos, proteínas y carbohidratos. Las propiedades catabólicas de los hidrocarburos, junto con las capacidades de producción de bio-surfactantes, es una ventaja que puede ser aprovechada para la limpieza de terrenos contaminados con petróleo y también en la industria alimentaria y cosmética.

Growth and biosurfactant synthesis by Nigerian hydrocarbon-degrading estuarine bacteria.

The ability of microorganisms to degrade petroleum hydrocarbons is important for finding an environmentally-friendly method to restoring contaminated environmental matrices. Screening of hydrocarbon-utilizing and biosurfactant-producing abilities of organisms from an estuarine ecosystem in Nigeria, Africa, resulted in the isolation of five microbial strains identified as Corynebacterium sp. DDV1, Flavobacterium sp. DDV2, Micrococcus roseus DDV3, Pseudomonas aeruginosa DDV4 and Saccharomyces cerevisae DDV5. These isolates grew readily on several hydrocarbons including hexadecane, dodecane, crude oil and petroleum fractions. Axenic cultures of the organisms utilized diesel oil (1.0% v/v) with generation times that ranged significantly (t-test, P < 0.05) between 3.25 and 3.88 day, with concomitant production of biosurfactants. Kinetics of growth indicates that biosurfactant synthesis occurred predominantly during exponential growth phase, suggesting that the bioactive molecules are primary metabolites. Strains DDV1 and DDV4 were evidently the most metabolically active in terms of substrate utilization and biosurfactant synthesis compared to other strains with respective emulsification index of 63 and 78%. Preliminary biochemical characterization indicates that the biosurfactants are heteropolymers consisting of lipid, protein and carbohydrate moieties. The hydrocarbon catabolic properties coupled with biosurfactant-producing capabilities is an asset that could be exploited for cleanup of oil-contaminated matrices and also in food and cosmetic industries.

Probiotics and zinc in acute infectious gastroenteritis in children: are they effective?

OBJECTIVE: Several studies in recent decades have assessed the effects of different probiotics in acute gastroenteritis, showing that specific strains of Lactobacilli (mainly Lactobacillus casei GG) and Saccharomyces boulardii may exert some beneficial therapeutic actions, mainly when used in rotavirus gastroenteritis, at a high dose, and in the early phase. The mechanisms of action of probiotics are not completely elucidated but seem to involve a complex interaction among epithelial, molecular, metabolic, and immune responses. Data on the prevention of community-acquired, nosocomial, and travelers' diarrhea are currently conflicting. Because each micro-organism has different properties, an accurate selection of the strain, dose, and patient should be cautiously considered. METHODS AND RESULTS: Several reports from developing countries have demonstrated that supplements of zinc also provide significant reduction in stool output and duration, persistency, and severity of diarrhea. In view of the published data and of the different actions of zinc (such as improvement of the immune status, intestinal permeability, epithelial and enzymatic function, and electrolyte transport), the use of zinc as adjunctive therapy to oral rehydration solution has the potential to improve the management of diarrhea and decrease complications in children worldwide. In contrast to probiotics, which most trials in the developed world have used, there has been no trial with zinc performed in developed countries. CONCLUSION: Data on the effect of a combined administration of zinc and probiotics in acute gastroenteritis are urgently needed.

Selenium enrichment and anti-oxidant status in baker’s yeast, Saccharomyces cerevisiae at different sodium selenite concentrations / Enriquecimiento con selenio y estado anti-oxidante de la levadura de harinas Saccharomyces Cerevisiae con diferentes concentraciones de selenito sódico

The use of selenized yeast as enriched selenium supplements in human nutrition has become a topic of increasing interest over the last decade. The present study was designed with the aim to achieve a balance between selenium (Se) incorporation and optimal growth of yeast cells along with effect of Se enrichment on antioxidant defense status of yeast cells. Since oxidative stress has been known to play a role in the life span of all types of cells, so in the present studies anti-oxidant defense status was evaluated in the Se- enriched baker’s yeast cell culture model. Upon Se supplementation as sodium selenite at various concentrations in the growth medium, a continuous increase in glutathione peroxidase (GSH-Px) activity and Se content was observed. In case of reduced glutathione (GSH) decreasing trend were observed with increasing Se concentrations An increasing trend in total glutathione as well as glutathione-s-transferase activity was observed at increasing Se concentrations. Thus, Se supplementation significantly enhanced GSH-Px levels along with alterations in other anti-oxidant enzymes, suggesting the role of Se in the enzyme defense system of yeast against oxidative damage. Further, as Se exerts growth inhibitory effect on cells, the growth inhibition study was carried out and decrease in biomass was observed with increasing concentrations of Se. Due to nutritional benefits, Se-enriched yeast may be considered a safe source of Se supplementation (AU)

El uso de levaduras “selenizadas” como suplementos enriquecidos con selenio en nutrición humana se ha convertido en un tema de interés creciente en la última década. Este estudio se diseño con el objetivo de conseguir un equilibrio entre la incorporación de selenio (Se) y el crecimiento óptimo de las células levaduriformes, junto con el efecto del enriquecimiento de Se sobre el estado de defensa anti-oxidante de las levaduras. Puesto que se sabe que el estrés oxidativo desempeña una función en la longevidad de todo tipo de células, en este estudio se determinó pues el estado de defensa anti-oxidante en un modelo de levadura de la harina enriquecida con selenio. Tras la complementación con Se, en forma de selenito sódico, en concentraciones variables en el medio de cultivo, se observó un aumento sostenido de la actividad glutatión peroxidasa (GSH-Px) y del contenido en Se. Con respecto al glutatión reducido (GSH), se observó una tendencia a la baja a medida que aumentaban las concentraciones de Se. Se observó una tendencia al alza del glutatión total y de la actividad glutatión-s-transferasa a medida que aumentaba la concentración de Se. Por lo tanto, la complementación con Se favoreció de forma significativa las concentraciones de GSH-Px junto con cambios en otras enzimas anti-oxidantes, lo que sugiere un papel del Se en el sistema enzimático de defensa de la levadura frente al daño oxidativo. Además, puesto que el Se ejerce un efecto inhibidor del crecimiento celular, se realizó un estudio de inhibición del crecimiento y se observó un descenso de la biomasa con concentraciones crecientes de Se. Dados los beneficios nutritivos, se podría considerar la levadura enriquecida con Se como una fuente segura de complementación de Se (AU)

Balance of flora, galt, and mucosal integrity.

It is clear that there is a dynamic relationship involving the gastrointestinal flora, environmental inputs (food and other nutrients), and the health of the immune system. Recent research has taught us a great deal about the role of diet and commensal bacteria in promoting health. It appears that Nobel Laureate Eli Metchnikov may have been correct in his assertion that live bacterial cultures are "the elixir of life". We are unlocking a number of secrets about immune system functioning, but we keep coming back to a simple intervention that has an ever-expanding opus of research to support it, and an extremely low toxicity ratio. Future studies will help us to clarify the best strains and the best dosages for individual patients and specific conditions. Assessment of commensal flora and a genomic scan for markers of immunologic dysregulation will be more accurate and more widely available. It appears, however, that the diagnostic and therapeutic tools we have to work with today can make a tremendous difference in reducing the burden of suffering for our patients. If "form follows function," as Buckminster Fuller was fond of saying, then the form of our immune system may be following the precise functions that our commensal flora is dictating. We have the opportunity to encourage breastfeeding, decrease unnecessary antibiotic and antimicrobial usage (especially in the first two years of life), improve oral tolerance with a healthy n-6/n-3 fatty acid ratio, and support the development of a healthy commensal flora. These actions on behalf of our immune systems will pay dividends for years to come.

Analysis of condensed and hydrolysable tannins from commercial plant extracts.

High performance liquid chromatography (HPLC)/DAD and MS qualitative and quantitative analyses of polyphenols, hydrolysable and condensed tannins from Pinus maritima L. and tannic acid (TA) extracts were performed using normal and reverse phase. Normal-phase HPLC was more suitable for pine bark (PBE) and tannic acid extracts analysis. The chromatographic profile revealed that P. maritima L. extract was mainly composed by polymeric flavanols (containing from two to seven units) and tannic acid (characterized by a mixture of glucose gallates containing from three to seven units of gallic acid). Concerning their antimycotic properties, P. maritima L. extract exhibited a broad activity towards yeast strains of the genera Candida, Cryptococcus, Filobasidiella, Issatchenkia, Saccharomyces: MICs from 200 to 4000 microg/ml (corresponding to 140-2800 microg/ml of active polyphenols) were determined. Conversely, no activity of tannic acid was observed over the same target microorganisms. Taken into consideration the above results of HPLC analysis and on the basis of the current literature, we may conclude that only 70.2% of polyphenols (recognized as condensed tannins) occurring in P. maritima L. extract can be apparently considered responsible for its antimycotic activity.

[Characterization of soy peptone for the culture of microorganisms]. / Caracterización de la peptona de soya para el cultivo de microorganismos.

A soy peptone obtained with enzyme papain as a hydrolyzing agent was characterized. The physicochemical evaluation of the nutritive base attained at pilot and industrial scale showed the following characteristics: loss due to desiccation, 1.93%; aminic nitrogen, 1.71%; total nitrogen, 8.63%; chlorides (as NaCl), 5.45%; and pH, 6.94. For the functional evaluation of the pilot and industrial batches (3), developed soy peptone and another one taken as a reference from the Biotécnica Internacional firm (México), was incorporated to a mixture of bases. There were no significant differences (p < 0.05) in the absorbance values at 640 nm obtained in the promotion of the growth between the developed product and that of reference for Staphylococcus aureus ATCC 25923. For Streptococcus pyogenes ATCC 19615, it was observed a mild superiority (significant difference for p < 0.05) starting from the 5th hour of incubation in favor of the industrial batches compared with the reference soy peptone. It was proved that the promotion of the growth in the soy triptone agar and broth media and in malt extract agar for the evaluated microorganisms was similar or higher (significant differences for p < 0.05) in those prepared with the experimental soy peptone, in comparison with the formulated with the reference base.

[The breeding and culture condition optimization of a high-biomass, selenium-enriched yeast strain].

The yeast fusant ZFF-28, which is high in biomass production and rich in selenium, was constructed after mutagenesis and protoplasts fusion between yeast strains. The total selenium content of ZFF-28 is 1.8 and 1.0 times higher than that of the parental strains Saccharomyces cerevisiae ZY-67 and Saccharomyces kluyveri SZY-198 respectively. Using single factor tests and a L16(4(3) x 2(1)) orthogonal design, the cultivation conditions was optimized as: 50mL culture in 250mL shake flasks in molasses containing 6% sugar and 60microg/mL Se at 28 degree C for 25h at 220 r/min, with the initial pH adjusted to 6.0 - 6.5. Under the optimized conditions, the biomass (dry weight) reached 8.2g/L and the Se content of the cells reached 2050microg/g, with organic and inorganic Se contents being 91% and 9% respectively.

Microbiological studies of feed and faeces of fatteners fed mixtures containing naked oat and permutite.

Quantitative and qualitative microbiological studies of feed mixtures and faeces of fatteners with body weight approximately 70 kg receiving complete mixtures: group I (control)--55% barley, II--55% naked oat, III--55% naked oat + 3% permutite were conducted. All mixtures were supplemented with the same protein concentrate for fatteners. It was found that feeding the fatteners with mixtures containing naked oat had a favourable effect on the composition of the faecal microflora, with increased participation of Lactobacillus spp. bacteria. The addition of permutite did not visibly affect the composition of the microflora and the observed differences between the groups can be attributed to the species of cereal in the feed mixture, with naked oat being far more favourable than barley. The addition of permutite seems to have a beneficial effect on the condition of the faecal matter of these animals, which contained a larger number of Saccharomyces spp. cells.

Influence of yeast flocculation on the rate of Jerusalem artichoke extract fermentation.

Variations in residual sugar composition have been observed during Jerusalem artichoke extract fermentations by using Saccharomyces diastaticus NCYC 625, a flocculating yeast strain. In batch cultures, these differences were due to the inulin polymer size distribution of the extracts: measurements of enzymatic activities on different polymerized substrates have shown that the hydrolysis and fermentation yield decreased when the fructose/glucose ratio of the extract increased. Inulin hydrolysis appeared to be the limiting factor of the fermentation rate. A comparison of continuous and batch cultures with the same extract showed that fermentability differences were related to the structure and size of the yeast flocs. This led to an hydrolysis selectivity of the inulin polymers according to their size: the chemostat culture in which the floc average size was larger gave longer chained residual sugars.

An attempt to use selected strains of bacteria adapted to high concentrations of petroleum oil to increase the effective removal of petroleum products in excess activated sludge in laboratory conditions.

Forty two strains of bacteria were isolated from excess activated sludge from petroleum wastewater treatment plant. The strains were identified and classified to the following groups: Enterobacteriaceae (7 strains), Anitratum (3 strains), Pseudomonas (13 strains), Micrococcus (12 strains), Comamonas (2 strains), Xanthomonas (2 strains), Achromobacter (1 strain) and Vibrio-Aeromonas (1 strain). One of the isolates was found to be a yeast strain. Following preliminary selection ten strains, showing the best growth in medium with oil fraction as sole carbon source, were chosen for further studies. The selected strains belong to Pseudomonas (6 strains), Xanthomonas (2 strains), Micrococcus (1 strain) and Saccharomyces (1 strain). The strains were adapted to high oil concentration (500-2000 mg/L) and an attempt to use them to intensify removal of petroleum products from excess activated sludge was made. The sludge was inoculated with a mixture of the isolated strains. The experiment was carried out three times, each time with a fresh sample of the excess sludge. The obtained results show that the inoculation of activated sludge with the strains active against oil reduced the petroleum products content by 20% in 14 days. The greatest reduction of oil was observed in sludge with the lowest dry weight, that is with the greatest degree of hydration. The dry weight of the excess sludge did not significantly decrease during the course of the experiment, after having been inoculated with the mixture of strains.

Selenoproteins in mitochondria and cytosol of Saccharomyces uvarum after growth in sodium selenite-supplemented media.

The extent of the biosynthesis of selenoproteins in mitochondria and cytosol of Saccharomyces uvarum depends on the sodium selenite concentration in the medium. In mitochondria there is a selenoprotein (SP 1) exhibiting glutathione peroxidase activity whose concentration already reaches a maximum at low concentrations of sodium selenite. A second selenoprotein (SP 2) was found in mitochondria and cytosol. Both proteins contain L-selenocysteine. The molecular masses of SP 1 and SP 2 are 73,000 Da and 83,000 Da, respectively. A subunit of SP 1 was found to have a molecular mass of 30,000 Da. SP 2, identified as a glycoprotein, has subunits with molecular masses of 36,500 Da and 5,000 Da. The selenium concentration of the total yeast increases from 260 micrograms/kg dry weight to 280 mg/kg dry weight after supplementation of the medium with sodium selenite.

Magnesium limitation and its role in apparent toxicity of ethanol during yeast fermentation.

The rate of ethanol production per milligram of cell protein begins to decline in the early stage of batch fermentation before high concentrations of ethanol have accumulated. In yeast extract-peptone medium (20% glucose), this initial decline appears to be related to growth and to result in part from a nutrient deficiency. The addition of yeast extract, peptone, and ashed preparations of these restored the ability of glucose-reconstituted medium (in which cells had been previously grown) to support vigorous growth. Magnesium was identified as the active component. Supplementing fermentations with 0.5 mM magnesium prolonged exponential growth, resulting in increased yeast cell mass. The addition of magnesium also reduced the decline in fermentative activity (micromoles of CO2 evolved per hour per milligram of protein) during the completion of batch fermentations. These two effects reduced the time required for the conversion of 20% glucose into ethanol by 1/3 with no measurable loss in ethanol yield (98% of theoretical maximum yield). It is possible that some of the reported beneficial effects of complex nutrients (soy flour and yeast extract) for ethanol production also result from the correction of a simple inorganic ion deficiency, such as magnesium.