Effect of culture media on the composition of free amino acids in Saccharomyces cerevisiae yeast

The quantitative and qualitative compositions of free amino acids of the yeast Saccharomyces cerevisiae Y-503 cultivated in different nutrient media were studied by liquid chromatography. The yeast grown in the medium containing geothermal water was shown to accumulate more amino acids. During lyophilization, the stabilization of the physiological activity of the yeast in this nutrient medium was observed. The increased biological value of dry yeast was shown to depend on the content of free amino acids, including essential amino acids: arginine, histidine, leucine, isoleucine, lysine, threonine, serine and phenylalanine.     

Comparison of fermentative capacities of industrial baking and wild-type yeasts of the species Saccharomyces cerevisiae in different sugar media

AIMS: To compare the fermentative capacity of wild and domesticated isolates of the genus Saccharomyces. METHODS AND RESULTS: The fermentative capacity of yeasts from a variety of wild and domesticated sources was tested in synthetic dough media that mimic major bread dough types. Domesticated yeast strains were found to have better maltose-utilizing capacity than wild yeast strains. The capacity to ferment sugars under high osmotic stress was randomly distributed amongst wild and baking strains of Saccharomyces. CONCLUSION: The domestication of bakers' yeast has enhanced the ability of yeasts to ferment maltose, without a similar impact on the fermentative capacity under high osmotic conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This study, combined with molecular studies of both wild and domesticated yeast, showed that domestication of bakers' yeast has resulted in improved maltose utilization, apparently via the duplication and mutation of the MAL genes.     

Biosynthesis of vitamins B by the fungus Pleurotus ostreatus in a submerged culture

The intra- and extracellular contents of vitamins were studied in the course of submerged cultivation of the higher basidial mushroom Pleurotus ostreatus (Jacq.: Fr.) Kummer st. IMBF-1300 on liquid nutrient media. This strain was found to be autotrophic in respect of thiamin (vitamin B1), riboflavin (vitamin B2), niacin (vitamin B5), pyridoxine (vitamin B6) and biotin (vitamin B7), but it failed to synthesize cyanocobalamin (vitamin B12). The composition and pH of the culture medium, containing such complex biostimulating supplements as maize extract and concentrated potato sap noticeably influence the contents of vitamins B1, B5 and B7 in the mycelium, and to a less degree they change the level of the intracellular biosynthesis of vitamins B2 and B6. Higher excretion of vitamins B5, B7 and especially B6 was observed on the semisynthetic media during the postexponential growth. Under experimental conditions vitamins B1 and B2 were accumulated only in the cells. The dry mycelium of P. ostreatus obtained by submerged cultivation on liquid media is a valuable source of B vitamins and, especially, of niacin. Thus the oyster mushroom and other edible mushrooms can be put at one of the top places among food-stuffs by the content of niacin.     

Utilization and stability of vitamins in serum-containing and serum-free media in CHO cell culture

The concentrations of four vitamins, ascorbic acid, nicotinamide, choline and thiamine were evaluated in the culture supernatant of Chinese hamster ovary (CHO) cells. The media used were -modified Eagle's minimum essential medium (MEM-) supplemented with 10% fetal calf serum, and a 1:1 mixture of Ham's F12 and Dulbecco's modified Eagle's medium (DME/F12), containing neither serum nor protein. The reference experiment without cells revealed instability of ascorbic acid and thiamine. Moreover, a significant amount of each vitamin decreased in the culture supernatant. The possibility of growth limitation by vitamin depletion is strongly suggested.     

Doublet cells inTetrahymena as indicators of culture media composition

Stomatogenesis in ciliates is a complex and carefully orchestrated event. The exo⁻ mutant SB255 ofTetrahymena thermophila has defects in mucocyst formation and docking and can also have one or two mouths. Three common culture media (proteose peptone, Medium 357, and yeast extract) were analyzed for total C, N, and inorganic elements and then tested for their effect on the number of mouths present in SB255. Cultures of SB255 grown in Medium 357 consisted of a mixed population of cells with either two mouths (doublet) or one mouth. Cultures from the same original stock grown in Medium 357 (SBm) and in 1% proteose peptone (SBpp) had different percentages of doublet cells in 1-, 2-, 3-, and 4-d-old cultures. When transferred to and grown in 1% yeast medium, both SBpp and SBm cultures had increased percentages of doublets over a 4-d culture period. When grown in 0.1, 0.5, or 1% yeast medium for 2 d, both SBpp and SBm cultures had more doublets in 1% than in either 0.1 or 0.5% yeast medium. Cultures of SBm grown in Medium 357. or 1% yeast medium for 2 d had a 10-fold increase in doublet cells compared to the inoculum. After 2 d in 1% proteose peptone, SBm cultures had percentages of doublet cells almost equal to that of the inoculum. Immunofluorescence and scanning electron microscopy (SEM) were used to examine cellular morphology of the doublet cells. These findings suggest that enriched media promote the growth of doublet cells. Furthermore, these doublets could prove to be a useful model system for the study of biological roles of trace elements.     

Light-dependent generation of reactive oxygen species in cell culture media

Cell culture media (RPMI 1640, Dulbecco’s Minimal Essential Medium and yeast extract-peptone-glucose medium) were found to oxidize dichlorodihydrofluorescein diacetate and dihydrorhodamine 123, and to generate spin adduct of 5,5′-dimethyl-1-pyrroline N-oxide, which indicates formation of reactive oxygen species (ROS). The production of ROS was light dependent. The main component of the media responsible for the generation of ROS was riboflavin, but tryptophan, tyrosine, pyridoxine, and folic acid enhanced the effect of riboflavin. These observations point to exposure of cells to ROS under in vitro culture conditions.     

Modification changes of the genetic material in Saccharomyces yeasts

The problem of mating-type switches in heterothallic yeast cells was investigated. In selective system for cytoduction in alpha x alpha crosses alpha-cytoductants were predominantly obtained. Thus matings in alpha x alpha crosses can proceed through non-heritable changes (modifications) of the mating type alpha----a. The frequency of alpha-cytoductants after UV-irradiation of the recipient cells exceeded the control value 50-90 times. The extra copy of MAT alpha dramatically decreased the frequency of cytoductants in alpha x alpha crosses, either spontaneously or after UV-irradiation. The rad18 recipient defective in postreplication repair had 70-times increased level of mating-type modifications, as compared with isogenic Rad+ strain. An explanation consistent with these data is that mating-type modifications are due to phenotypic expression of primary lesions of MAT alpha locus. Such lesions might be expressed as transient a-mating type. After the mating event, these lesions can be repaired or turned to true mutations within the MAT locus. In fact, approximately half of non-mating cytoductants from alpha x alpha crosses had the phenotype of mat alpha 2 mutants.     

Molecular genetic diversity of the Saccharomyces yeasts in Taiwan: Saccharomyces arboricola, Saccharomyces cerevisiae and Saccharomyces kudriavzevii

Genetic hybridization, sequence and karyotypic analyses of natural Saccharomyces yeasts isolated in different regions of Taiwan revealed three biological species: Saccharomyces arboricola, Saccharomyces cerevisiae and Saccharomyces kudriavzevii. Intraspecies variability of the D1/D2 and ITS1 rDNA sequences was detected among S. cerevisiae and S. kudriavzevii isolates. According to molecular and genetic analyses, the cosmopolitan species S. cerevisiae and S. kudriavzevii contain local divergent populations in Taiwan, Malaysia and Japan. Six of the seven known Saccharomyces species are documented in East Asia: S. arboricola, S. bayanus, S. cerevisiae, S. kudriavzevii, S. mikatae, and S. paradoxus.     

Efficacy of agar media for enumerating two Saccharomyces species in sucrose syrups

Experiments were carried out to determine the suitability of acidified and antibiotic-supplemented agars for supporting colony formation by cells ofSaccharomyces cerevisiae andSaccharomyces rouxii. Yeasts had been suspended in sucrose syrups buffered at pH 3.0, 5.0 and 7.0, and stored at 4 and 21°C for periods of time ranging to 10 weeks. TheSaccharomyces species were recovered in equal numbers, regardless of the type of enumeration agar. Sucrose, at concentrations up to 60%, protected cells against inactivation during storage, and survival was greater at 21° than at 4°C.     

Efficacy of agar media for enumerating two Saccharomyces species in sucrose syrups

Experiments were carried out to determine the suitability of acidified and antibiotic-supplemented agars for supporting colony formation by cells ofSaccharomyces cerevisiae andSaccharomyces rouxii. Yeasts had been suspended in sucrose syrups buffered at pH 3.0, 5.0 and 7.0, and stored at 4 and 21°C for periods of time ranging to 10 weeks. TheSaccharomyces species were recovered in equal numbers, regardless of the type of enumeration agar. Sucrose, at concentrations up to 60%, protected cells against inactivation during storage, and survival was greater at 21° than at 4°C.     

An overview on glutathione in Saccharomyces versus non-conventional yeasts

Glutathione (GSH: L-gamma-glutamyl-L-cysteinylglycine) is present in high concentrations up to 10 mM in yeast cells. Its very low redox potential (E'(o)=-240 mV for thiol disulfide exchange) gives this tripeptide the properties of a cellular redox buffer. In Saccharomyces cerevisiae and non-conventional yeasts (NCY), GSH may be involved in basic cellular functions such as the maintenance of mitochondrial and membrane integrity. GSH also assumes pivotal roles in (i) response to sulfur and nitrogen starvation; (ii) detoxification of endogenous toxic metabolites, such as excess formaldehyde produced during the growth of the methylotrophic yeasts Hansenula polymorpha, Candida boidinii and Kloeckera sp.; (iii) protection against oxidative stress provoked by exposure of the cells to reactive oxygen species including peroxides and hydroperoxides; (iv) detoxification of xenobiotics such as halogenated aromatics, alkylating agents and arsenite; (v) resistance to heavy-metal stress exemplified by the responses of S. cerevisiae and Schizosaccharomyces pombe to cadmium salts; (vi) yeast<-->mycelium transition in Candida and Aureobasidium sp.     

Genetic differentiation of the sherry yeasts Saccharomyces cerevisiae

Molecular genetic study of the yeast Saccharomyces cerevisiae isolated at various stages of sherry making (young wine, solera, and criadera) in various winemaking regions of Spain demonstrated that sherry yeasts diverged from the primary winemaking yeasts according to several physiological and molecular markers. All sherry strains independently of the place and time of their isolation carry a 24-bp deletion in the ITS 1 region of ribosomal DNA, whereas the yeasts of the primary winemaking lack this deletion. Molecular karyotypes of the sherry yeast from different populations were found very similar.     

B vitamins in the nervous system

The coenzyme functions of the B vitamins in intermediatry metabolism are well established; nevertheless, for none of them is it possible to determine precisely the connection between the biochemical lesions associated with deficiency and the neurological consequences. Although there is convincing evidence of a neurospecific role for thiamin and other B vitamins, in no case has this role been adequately described. Similarly, the neurochemical sequelae of intoxication by massive amounts of vitamins (so-called mega-vitamin therapy or orthomolecular medicine) remain largely unexplained.