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PubMedDate not supplied

[Cultivation of Candida tropicalis yeasts on coal substrates].

The optimum conditions have been established in order to obtain aquaeous extracts of oxidized coal. Coal substrates seem to consist of aromatic polycarboxylic acids and unsaturated aliphatic compounds. Candida tropicalis was adapted to coal extracts, and about 2 g of absolutely dry yeast biomass had been obtained from 600 ml of the substrate in the conditions of a laboratory fermenter. The amino acid composition of the biomass was determined.

Amino Acids↗
PubMed1993

The principle of parabolic feed for a fed-batch culture of baker's yeast.

A parabolic nutrient supplementation has been derived by the specific growth rate, sugar conversion rate, and Pasteur effect, followed by high qualitative and quantitative biomass production with the lowest sugar consumption. The method produced about 95% cells in the G1 phase that were more resistant to drying and aging. These features are particularly important in the process of making dry yeast. It appears that the parabolic feed method may be used to species culture that show the Pasteur effect or produce byproducts from sugar. This may be because the supplementation is in conformity with reproducible kinetic growth during the fed-batch culture.

Carbohydrate Metabolism↗
PubMed1984

The purification of yeast glucose 6-phosphate dehydrogenase by dye-ligand chromatography.

Glucose 6-phosphate dehydrogenase (EC 1.1.1.39) has been purified to homogeneity from baker's yeast by a simple procedure involving affinity elution from a column of red triazine dye, H-8BN, immobilized to Sepharose 6B. Eight milligrams of homogeneous protein is obtained in 53% yield from 200 g of dried yeast. This represents the first published purification of the enzyme from Saccharomyces Cerevisiae.

Chromatography↗
PubMed2005

Fine measurement of ergosterol requirements for growth of Saccharomyces cerevisiae during alcoholic fermentation.

Yeasts can incorporate a wide variety of exogenous sterols under strict anaerobiosis. Yeasts normally require oxygen for growth when exogenous sterols are limiting, as this favours the synthesis of lipids (sterols and unsaturated fatty acids). Although much is known about the oxygen requirements of yeasts during anaerobic growth, little is known about their exact sterol requirements in such conditions. We developed a method to determine the amount of ergosterol required for the growth of several yeast strains. We found that pre-cultured yeast strains all contained similar amounts of stored sterols, but exhibited different ergosterol assimilation efficiencies in enological conditions [as measured by the ergosterol concentration required to sustain half the number of generations attributed to ergosterol assimilation (P(50))]. P(50) was correlated with the intensity of sterol synthesis. Active dry yeasts (ADYs) contained less stored sterols than their pre-cultured counterparts and displayed very different ergosterol assimilation efficiencies. We showed that five different batches of the same industrial Saccharomyces cerevisiae ADY exhibited significantly different ergosterol requirements for growth. These differences were mainly attributed to differences in initial sterol reserves. The method described here can therefore be used to quantify indirectly the sterol synthesis abilities of yeast strains and to estimate the size of sterol reserves.

Alcohols↗
PubMed1997

Yeast treatment to reduce Salmonella and Campylobacter populations associated with broiler chickens subjected to transport stress.

The prevalence of Campylobacter and Salmonella on retail poultry carcasses remains a significant public health concern. The stresses associated with transporting poultry prior to slaughter have been shown to increase pathogen populations both in the intestinal tract and on the carcass exterior. The yeast, Saccharomyces boulardii, was evaluated for its ability to reduce populations of Salmonella and Campylobacter in broiler chickens subjected to transport stress. Chicks, inoculated with individual strains of Salmonella and Campylobacter were held for 6 wk and then divided into two groups with half of the chickens receiving 10% dried yeast in the feed for 60 h. The birds were then caged and transported to simulate commercial conditions. After euthanatizing the birds, the ceca were aseptically removed and analyzed for Salmonella and Campylobacter. With no yeast treatment, transport stress caused the Salmonella colonization frequency to increase more than fivefold, from 3.3 to 16.7%. Yeast treatment significantly reduced the frequency of Salmonella colonization to lower than prestress levels, as no Salmonella were recovered from the ceca of these birds (P < 0.05). Similar results were obtained from birds challenged with a mixture of Salmonella and Campylobacter strains. Before transport, 53.3% of these chickens were positive for Salmonella. Transport stress increased the colonization rate to 67.5% in control birds, whereas the colonization of yeast-treated chickens decreased to 40% (P < 0.05). Frequency of Campylobacter isolation from the ceca was not affected by treatment, but Campylobacter populations present in the ceca were significantly reduced in the mixed strain trial (P < 0.05).

Analysis of Variance↗
PubMed1984

Identification of 17 beta-estradiol as the estrogenic substance in Saccharomyces cerevisiae.

Saccharomyces cerevisiae possesses a high-affinity estrogen binding protein and an endogenous ligand that displaces [3H]estradiol from both the yeast binding protein and mammalian estrogen receptors. Semipurified preparations of this ligand have been shown to exhibit estrogenic activity in mammalian systems. We now describe the purification procedure and ultimate identification of the estrogenic substance in extracts of S. cerevisiae as 17 beta-estradiol. Organic solvent extracts of commercially obtained dried yeast were sequentially chromatographed on silica gel columns and then subjected to a series of reversed phase HPLC fractionations. Active ligand was monitored by [3H]estradiol displacement in a rat uterine cytosol assay. After seven chromatography steps, the purified and highly active ligand exhibited a single peak with retention times identical to those of 17 beta-estradiol on both HPLC and GC. The yeast material was identified as 17 beta-estradiol by its UV absorbance and mass spectrometric fragmentation pattern. In addition, radioimmunoassay confirmed the presence of approximately the same mass of 17 beta-estradiol (approximately equal to 800 ng/1.5 kg of yeast) as estimated both by a competitive binding assay with estrogen receptor and by mass spectrometry. Extraneous contamination by estradiol was excluded by repeat experiments with different batches of starting material and demonstration of estradiol by RIA in conditioned medium and cell pellets of laboratory-grown S. cerevisiae whereas non-conditioned medium did not possess the steroid. We conclude that 17 beta-estradiol is a yeast product.

Estradiol↗
PubMed1988

Water stress plating hypersensitivity of yeasts: protective role of trehalose in Saccharomyces cerevisiae.

Water stress plating hypersensitivity was studied in two strains of Saccharomyces cerevisiae, one of them being a mutant incapable of accumulating trehalose to significant levels. The wild-type strain was grown in a defined medium with glucose, maltose or ethanol as carbon/energy source. In each case plating hypersensitivity was demonstrated and resistance to the stress developed in the second half of the exponential growth phase. Development of resistance was accompanied by accumulation of trehalose and was apparently unrelated to glycerol content which, under these conditions, was always low. A qualitatively similar trend was observed in the mutant grown on glucose but trehalose levels remained low and recovery of stress resistance was only slight. Dinitrophenol induced trehalose breakdown in resting yeast and simultaneously induced the onset of plating hypersensitivity. A negative correlation was demonstrated between trehalose content and 'plating discrepancy' (log colony count on 'normal' agar-log colony count on stressing agar) for both strains under all experimental conditions. The correlation held for trehalose contents up to about 50 mg (g dry yeast)-1, above which the yeasts were apparently fully resistant. Trehalose was evidently a more effective compatible solute, per mole, than glycerol.

Colony Count, Microbial↗
PubMed2005

Influence of temperature and pH on Saccharomyces bayanus var. uvarum growth; impact of a wine yeast interspecific hybridization on these parameters.

The species Saccharomyces bayanus var. uvarum possesses interesting enological characteristics but produces high concentration of volatile fermentative compounds not desirable in Sauvignon blanc wines. Interspecific hybrids between Saccharomyces cerevisiae and S. bayanus var. uvarum were made in order to join the main parental advantages. Two hybrids were selected on the basis of their fermentation characteristics and their karyotypes, i.e. they have a different mitochondrial DNA. In order to produce these hybrids as active dry yeast to be used as starter in wine-making, their optimal environmental conditions for growth, i.e. temperature and pH, were determined as the objective of our work. Using a two-level factorial design it was found that the two parental strains have different optimal temperature but for the two strains, pH does not have a significant influence on growth. The influence of temperature on biomass productivity for hybrid strains were strictly identical, so we suppose that the main genes coding for temperature sensitivity were not contained in mitochondrial DNA, but in nuclear DNA. Moreover the reactions of hybrid strains to the temperature variations were similar to the one of S. bayanus var. uvarum. This latter strain could have a majority of genes responsible of temperature sensitivity dominant in comparison with those of the strain S. cerevisiae.

Biomass↗
PubMed1994

Effect of moisture content on the invertase activity of freeze-dried S. cerevisiae.

The invertase activity of intact Saccharomyces cerevisiae submitted to freezing-thawing was affected by pH, the chemical nature of the buffer, and the freezing cooling rate (CR), leading in some cases to a complete invertase inactivation (acetate buffer, pH 4.0, CR = 0.5 degree C/min). Once established under adequate freezing conditions the invertase activity remained unchanged after freeze-drying. Nevertheless, in some cases the cell-growing capability after freeze-drying diminished around 70%, mainly if the frozen cell suspension was attained through freezing carried out at CR = 0.5 degree C/min. Water sorption isotherms of freeze-dried samples (freeze-dryer Edwards L-4KR; 30 degrees C and 0.1 mB) were determined at 10 and 25 degrees C. The monolayer moisture content (MMC) at each temperature (12.7 and 3.71 for 10 and 25 degrees C, respectively) was calculated from isotherms by applying BET and GAB models. Freeze-dried yeast with water activity (Aw) between 0 and 0.33 (about the MMC value) maintained at 25 degrees C for 235 days and at 89 degrees C for 15 min retained at least 85% of its original invertase activity (IA), whereas samples with Aw > MMC lost at least 60% of its IA. X ray diffraction showed that the freeze-dried cake before and after storage presented an amorphous structure.

Buffers↗
PubMed1996

Biotransformation of uridine monophosphate (UMP) and glucose to uridine diphosphate-glucose (UDPG) by Candida saitoana KCTC7249 cells.

The present study investigates the biotransformation of glucose with uridine monophosphate (UMP) to obtain sugar nucleotide, UDP-glucose (UDPG), by the dried cells of Candida saitoana KCTC7249. The biotransformation was optimized by varying the concentrations of substrates and phosphate ion. UDPG (24 mM) was biotransformed from 200 mM glucose and 37.5 mM UMP by dried cells of C. saitoana. The glucose yields about 64% UDP-glucose, based on UMP concentration. The addition of glucose-1-phosphate to the reaction mixture accelerated the formation of UDPG from a concentration of UMP. The structure of UDP-glucose obtained was determined with 13C NMR and FAB mass spectra. These results indicate that the yeast-dried cells could be used for the production of nucleotide sugars for donor molecules of complex carbohydrate synthesis.

Candida↗
PubMed1977

Lipid accumulation in an oleaginous yeast (Candida 107) growing on glucose under various conditions in a one- and two-stage continuous culture.

Lipid accumulation and fatty acid composition in Candida 107 have been studied using a two-stage continuous culture system in which the first vessel was run under carbon-limited conditions and then the entire output was passed into a second vessel, where lipid accumulation was stimulated by adding only glucose. Maximum lipid accumulation (28% of yeast [dry weight]) occurred for a volume ratio of vessel 1 to vessel 2 of 3:5, with 30 g of glucose per liter being added to vessel 2 operated at 25 degrees C with an aeration rate of between 0.1 and 1.0 volume of air/volume of medium per min. Although the maximum specific rate of lipid formation (0.05 g of lipid/g of yeast per h) was higher than in a nitrogen-limited, single-stage system, the efficiency of lipid formation was much less and never exceeded 14 g of lipid produced per 100 g of glucose consumed. The fatty acid composition was not significantly altered in either the two-stage or single-stage culture (nitrogen-limited) systems by changes in growth temperature (from 19 to 33 degrees C) or aeration rates (0.05 to 1.0 volume of air/volume of medium per min); or, in the two-stage system, by changes in the residence time of the yeast in the second vessel (from 3.2 to 24.4 h), or, in the single-stage system, by changes in pH (from 3.5 to 7.5). Only when the concentration of glucose entering vessel 2 of the two-stage system was less than 30 g/liter did significant changes in the fatty acids occur. Thus, although a two-stage continuous culture system allows lipid accumulation to be separated from the growth phase, it offers no practical advantages over a single-stage system as a means of producing microbial oils and fats.

Journal Article↗
PubMed1989

Improved representation of cell surface structures by freeze substitution and backscattered electron imaging.

A surface preparation method for the SEM based on cryofixation is presented suitable for the demonstration of membrane particles on whole cells. LLC-PK1 cells (a renal epithelial cell line in culture) were fast frozen, freeze substituted, critical point dried, shadowed with 2 nm of platinum carbon and stabilized with a carbon backing layer. Membrane bound particles are visualized by the material dependent backscattered electron image mainly originating from the platinum shadow. The surface of the LLC-PK1 cells is almost free of precipitated material indicating that the culture medium is removed during freeze substitution or critical point drying. The apical plasma membrane with microvilli and ciliae is well preserved and differences in particle density can be detected. The feasibility of the coating technique for backscattered electron imaging was tested on the well known hexagonally arranged intramembranous particles of fractured and partially freeze dried yeast. This 16.5 nm periodic structure is clearly demonstrable on the bulk SEM-specimen stablized with the carbon backing layer. Without a carbon layer severe shrinking artifacts occurred.

Animals↗
PubMed1983

A rapid procedure for the preparation of highly purified pyruvate decarboxylase from brewer's yeast.

A rapid purification procedure for pyruvate decarboxylase (E.C. 4.1.1.1.) from fresh cells of brewer's yeast (Saccharomyces carlsb.) is reported. The preparation of a crude enzyme (30-45 U/mg) by the use of fractionation steps with protamine sulfate, acetone, and ammonium sulfate takes about 6-7 h. A stable pyruvate decarboxylase (70-85 U/mg) was obtained from such preparations after purification on CM Sephadex C 50 after another 2-3 h. Stability and structural properties are compared for enzymes prepared from fresh and dried yeast.

Carboxy-Lyases↗
PubMed2004

Comparative analysis of trehalose production by Debaryomyces hansenii and Saccharomyces cerevisiae under saline stress.

The comparative analysis of growth, intracellular content of Na+ and K+, and the production of trehalose in the halophilic Debaryomyces hansenii and Saccharomyces cerevisiae were determined under saline stress. The yeast species were studied based on their ability to grow in the absence or presence of 0.6 or 1.0 M NaCl and KCl. D. hansenii strains grew better and accumulated more Na+ than S. cerevisiae under saline stress (0.6 and 1.0 M of NaCl), compared to S. cerevisiae strains under similar conditions. By two methods, we found that D. hansenii showed a higher production of trehalose, compared to S. cerevisiae; S. cerevisiae active dry yeast contained more trehalose than a regular commercial strain (S. cerevisiae La Azteca) under all conditions, except when the cells were grown in the presence of 1.0 M NaCl. In our experiments, it was found that D. hansenii accumulates more glycerol than trehalose under saline stress (2.0 and 3.0 M salts). However, under moderate NaCl stress, the cells accumulated more trehalose than glycerol. We suggest that the elevated production of trehalose in D. hansenii plays a role as reserve carbohydrate, as reported for other microorganisms.

Ascomycota↗
PubMedDate not supplied

[The viability assessment of ethanol-producing yeast by computer-aided fluorescence microscopy].

Vital staining of the ethanol-producing yeast Saccharomyces cerevisiae with ethidium bromide and DAPI allows intact and damaged cells to be differentiated by fluorescence microscopy. A computer image analysis procedure has been developed for the automatic determination of the relative number of damaged cells using ImageJ software (National Institute of Health, United States; http://rsb.info.nih.gov./ij/). A good correlation has been found between the viability rates determined by the plate count method and the relative numbers of intact cells assessed by the developed procedure in the dry-yeast preparations rehydrated under various conditions.

Colony Count, Microbial↗
PubMed2006

Interactions between Brettanomyces bruxellensis and other yeast species during the initial stages of winemaking.

AIMS: Wine is the product of complex interactions between yeasts and bacteria in grape must. Amongst yeast populations, two groups can be distinguished. The first, named non-Saccharomyces (NS), colonizes, with many other micro-organisms, the surface of grape berries. In the past, NS yeasts were primarily considered as spoilage micro-organisms. However, recent studies have established a positive contribution of certain NS yeasts to wine quality. Amongst the group of NS yeasts, Brettanomyces bruxellensis, which is not prevalent on wine grapes, plays an important part in the evolution of wine aroma. Some of their secondary metabolites, namely volatile phenols, are responsible for wine spoilage. The other group contributing to wine aroma, which is also the main agent of alcoholic fermentation (AF), is composed of Saccharomyces species. The fermenting must is a complex microbial ecosystem where numerous yeast strains grow and die according to their adaptation to the medium. Yeast-yeast interactions occur during winemaking right from the onset of AF. The aim of this study was to describe the interactions between B. bruxellensis, other NS and Saccharomyces cerevisiae during laboratory and practical scale winemaking. METHODS AND RESULTS: Molecular methods such as internal transcribed spacer-restriction fragment length polymorphism and polymerase chain reaction and denaturing gradient gel electrophoresis were used in laboratory scale experiments and cellar observations. The influence of different oenological practices, like the level of sulphiting at harvest time, cold maceration preceding AF, addition of commercial active dry yeasts on B. bruxellensis and other yeast interactions and their evolution during the initial stages of winemaking have been studied. Brettanomyces bruxellensis was the most adapted NS yeast at the beginning of AF, and towards the end of AF it appeared to be more resistant than S. cerevisiae to the conditions of increased alcohol and sugar limitation. CONCLUSIONS: Among all NS yeast species, B. bruxellensis is better adapted than other wild yeasts to resist in must and during AF. Moreover, B. bruxellensis appeared to be more tolerant to ethanol stress than S. cerevisiae and after AF B. bruxellensis was the main yeast species in wine. SIGNIFICANCE AND IMPACT OF THE STUDY: Brettanomyces bruxellensis interacts with other yeast species and adapts to the wine medium as the dominant yeast species at the end of AF. Contamination of B. bruxellensis might take place at the beginning of malolactic fermentation, which is a critical stage in winemaking.

DNA, Fungal↗
PubMed1996

Induction of the d-Amino Acid Oxidase from Trigonopsis variabilis.

Induction of the d-amino acid oxidase (EC. 1.4.3.3) from the yeast Trigonopsis variabilis was investigated by using a minimal medium containing glucose as the carbon and energy source, (NH(inf4))(inf2)SO(inf4) as the nitrogen source, and various d- and dl-amino acid derivatives as inducers. The best new inducers found were N-carbamoyl-d-alanine, N-acetyl-d-tryptophan, and N-chloroacetyl-d-(alpha)-aminobutyric acid; when the induction effects of these compounds were compared with the effects of d-alanine as the nitrogen source and inducer, the resulting activities of d-amino acid oxidase per gram of dried yeast were 4.2, 2.1, and 1.5 times higher, respectively. The optimum concentration of the best inducer, N-carbamoyl-d-alanine, was 5 mM. This inducer could also be used in its racemic form. The induction was pH dependent. After cultivation of the yeast in a 50-liter bioreactor, d-amino acid oxidase activity of about 3,850 (mu)kat (231,000 U) was obtained. In addition, production of the d-amino acid oxidase was found to be significantly dependent on the metal salt composition of the medium. Addition of zinc ions was required to obtain high d-amino acid oxidase levels in the cells. The optimum concentration of ZnSO(inf4) was about 140 (mu)M.

Journal Article↗