Showing papers on "Yeast published in 1975"

An amino acid liquid synthetic medium for the development of mycelial and yeast forms of Candida Albicans

Abstract: A chemically defined medium composed of 6 amino acids, biotin, inorganic salts and glucose for the growth of yeast and mycelial phases of Candida albicans at 25 degrees C and 37 degrees of C respectively was developed based on the aminopeptidase(s) profile of the fungus. This medium has proved successful in maintaining the growth characteristics of both phases during serial transfers. The relative pathogenicity, virulence, invasiveness and immunogenicity of the yeast and mycelial phases are discussed.

Genetic control of cell size at cell division in yeast

Abstract: A temperature-sensitive mutant strain of the fission yeast Schizosaccharomyces pombe has been isolated which divides at half the size of the wild type. Study of this strain suggests that there is a cell size control over DNA synthesis and a second control acting on nuclear division.

A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses

Abstract: 4′-6-DIAMIDINO-2-PHENYLINDOLE (DAPI), which was first synthesised by Dann et al.1 as a trypanocide related to Berenil, has been shown to possess useful DNA binding properties2. Thus, DAPI will bind differentially to yeast mitochondrial and nuclear DNA forming highly fluorescent complexes and enhancing the separation of the two DNAs in caesium chloride gradients2. DAPI can also be used as a highly specific fluorescent stain for both nuclear and mitochondrial DNA in yeast. It seems to be highly sensitive and probably permits detection of a single yeast mitochondrial DNA molecule (D.H.W., and D. J. Fennell, unpublished).

3 Alcohol Dehydrogenases

Abstract: Publisher Summary This chapter describes the advances with an emphasis on the structures of the alcohol dehydrogenases and the relationship between structure and function Yeast and mammalian alcohol dehydrogenase differ in substrate specificity and rate of catalytic activity The classic yeast enzyme is more specific for acetaldehyde and ethanol, which is consistent with its recognized physiological Significance to participate in alcohol fermentation at the end of the glycolytic pathway Enzyme forms with other functions and properties also occur in yeast The mammalian enzymes have broad substrate specificity and, even with primary alcohols, the maximum activity is not observed with ethanol Alcohols including ethanol, produced in the intestinal tracts mainly by bacterial actions, are found in the portal vein One physiological function of liver alcohol dehydrogenase may be to metabolize these products Structural studies have established that mammalian alcohol dehydrogenases have a distant evolutionary link to both the yeast and bacterial enzymes Ingested alcohol is metabolized to acetaldehyde mainly by the action of liver alcohol dehydrogenase

Adenylate energy charge in Saccharomyces cerevisiae during starvation.

Abstract: Bakers' yeast cells, Saccharomyces cerevisiae, if grown aerobically on ethanol or if grown aerobically on glucose and allowed to pass into stationary phase, with utilization of accumulated ethanol, maintain a normal value (0.8 to 0.9) of the adenylate energy charge during prolonged starvation. In contrast, cells grown anaerobically on glucose and cells in the early stages of aerobic growth on glucose exhibit a rapid decrease of energy charge if transferred to medium lacking on energy source. These results suggest that functional mitochondria or enzymes of balance of adenine nucleotides during starvation. Yeast cells remain viable at energy charge values below 0.1, in marked contrast to results previously obtained with Escherichia coli. In other respects, the engery charge responses of yeast to starvation and refeeding are generally similar to those previously reported for E. coli.

The occurrence of killer character in yeasts of various genera.

Abstract: Species of 7 of the 28 yeast genera in the National Collection of Yeast Cultures exhibited killing activity againstSaccharomyces cerevisiae. The highest incidence of killer yeasts was found in the genusHansenula (12 of the 29 strains examined).Saccharomyces, the best represented genus in the Collection, showed a low incidence of killer activity and many of the killer strains are hybrids with a commonS. cerevisiae parent. The activities of culture filtrates of the 59 killer yeast isolated responded differently to pH and four types of response were recognised.

Preparation of RNA and ribosomes from yeast.

Abstract: Publisher Summary This chapter discusses the preparation of RNA and ribosomes from yeast. Most methods used for the preparation of RNA and ribosomes from extracts of mammalian cells or bacteria can be directly applied either to yeast spheroplasts or to disrupted yeast cells. Yeast RNA can be radioactively labeled in vivo with purine and pyrimidine bases, phosphate- 32 P, and methyl-labeled methionine. Yeasts are able to utilize purines and pyrimidines added to the medium, although the free bases are not normal intermediates in the biosynthesis of nucleotides. Uracil and adenine are the bases most commonly used to label RNA. It is difficult to evaluate the usefulness for preparative purposes of the various labeling protocols, because the specific activities of the labeled RNAs are rarely determined. In the chapter, preparation of ribosomal subunits and polyribosomes and preparation of the high-molecular-weight rRNAs and low-molecular-weight rRNAs are discussed. An overview of polyacrylamide gel electrophoresis of low-molecular-weight RNAs and high-molecular-weight rRNA precursors is also presented in the chapter.

The stoicheiometry of the absorption of protons with phosphate and L-glutamate by yeasts of the genus Saccharomyces.

Abstract: 1. A study was made of the pH changes occurring when 0.1-4 mumol of glutamate, phosphate and certain phosphate esters was added at about pH 4.8 to washed cell preparations (50 mg dry wt.) of strains of Saccharomyces. The system also contained deoxyglucose and antimycin to inhibit energy metabolism and so prevent proton ejection from the yeast. 2. A strain of Sacc. carlsbergensis was grown in a chemostat with a limiting supply of phosphate in order to enhance the subsequent rate of phosphate transfer into the yeast. These preparations absorbed 0.2 mumol of phosphate with about 3 equiv. of protons/mol of phosphate. The charge balance was maintained by the efflux of 2 equiv. of K-+ from the yeast. 3. Larger amounts of phosphate were absorbed with fewer proton equivalents. 4. Arsenate and phosphate caused similar pH changes. 5. Glucose 6-phosphate, ATP and certain order phosphate esters each initiated a rise in pH, possibly because hydrolytic extracellular enzymes released phosphate that was subsequently absorbed. 6. Four strains of yeast were grown with glutamate as principal source of nitrogen. Each absorbed extra protons in the presence of L-glutamate. 7. One of them, a strain of Sacc. cerevisiae, absorbed 0.2 mumol of glutamate with 3equiv. of protons/mol of glutamate, and in these circumstances 1-2 equiv. of K-+ left the yeast cells. 8. The role of ionic gradients in the transport of these anions is discussed.

Isolation and chemical characterization of plasma membranes from the yeast and mycelial forms of Candida albicans.

Abstract: SUMMARY: It has been possible to induce the yeast-mycelium transformation in Candida albicans by growth of the organism under completely defined conditions in batch culture. Protoplasts have been obtained from the two forms by using a lytic enzyme preparation from Streptomyces violaceus. A plasma membrane fraction was prepared by osmotic lysis of these protoplasts and fractionated by using a combination of differential and discontinuous sucrose density-gradient flotation centrifugation. The purity of this fraction was determined by radioactive dansylation and iodination of plasma membranes of intact protoplasts followed by localization of the radioactivity upon fractionation. This procedure demonstrated less than 4% contamination of the plasma membrane fraction with other cell membranes. Chemical analysis of this fraction revealed that the major components were protein and lipid. Membranes from the yeast form contained (w/w): 50% protein, 45% lipid, 9% carbohydrate and 0.3% nucleic acid. Plasma membranes from the mycelial form contained significantly more carbohydrate and were found to be composed of (w/w): 43% protein, 31% lipid, 25% carbohydrate and 0.5% nucleic acid. Marked differences were also observed between the phospholipid, free and esterified sterols, and total fatty acids of membranes from the two forms of the organism.

Analytical methods for yeasts.

Abstract: Publisher Summary This chapter describes analytical methods for yeasts. Yeast cells present particular difficulties in the application of standard extractive and analytical methods. The thick and chemically refractile cell wall of the yeast cell often has a decided propensity to behave as a molecular filter, allowing only particular sizes of molecules in or out even when cell membrane integrity has been destroyed completely. Moreover, it might reasonably be suspected that the properties of the cell wall, like those of the membranes of the organism, may alter with the physiological status of the cell. Analytical methods applied to yeasts must provide a carefully struck balance between yield and reproducibility, together with reasonable facility. The methods of quantitative analysis of the major components of yeast cells described in the chapter have been selected because they provide a workable combination of these requirements. Quantitative assay of the protein content and RNA of whole cells and subcellular fractions is described in the chapter. Detection of specific proteins in yeast cells and identification of specific RNA components in yeast cells are also described in the chapter.

The nature of the stimulation of the growth of Streptococcus lactis by yeast extract

Abstract: Yeast extract was fractionated on Sephadex G-25 into 7 fractions. The fraction most stimulatory to the growth of Streptococcus lactis C10 contained over 70% of the amino N present in yeast extract and consisted of a wide variety of free amino acids and a small amount of peptide material. Examination of possible replacement factors for this fraction revealed that the amino -acid material present was largely responsible for the stimulation of Str. lactis C10. Purine and pyrimidine bases and inorganic constituents also contributed to the stimulation. In addition, yeast extract contained a component which decomposed H2O2, a metabolite which accumulates in the growth medium under aerobic conditions and inhibits growth. The nature of the stimulation was studied by isolating slow and fast acid-producing colonies of Str. lactis C10. It appeared that yeast extract and other amino-acid supplements prevented an observed inhibition of the growth of the slow variants below pH 6.0, apparently by satisfying a nutritional deficiency caused by a drop in pH.

Chapter 7 Methods for Monitoring the Growth of Yeast Cultures and for Dealing with the Clumping Problem

Abstract: Publisher Summary This chapter discusses methods for monitoring the growth of yeast cultures. Most biochemical, physiological, cytological, and developmental studies of yeasts require monitoring the growth of yeast cultures. In some cases, determination of the rate or extent of growth provides important experimental results; in many other cases, it is necessary to know the amount of cellular material analyzed or the physiological state of the cells, or both. The methods used for determining amounts of yeast and for monitoring the growth of yeast cultures are the same as those used with other microorganisms. This chapter focuses on the virtues and limitations of (and the relationships among) various methods of monitoring growth. It discusses the complications imposed by the budding mode of reproduction and by the clumping problem. The chapter describes the clumping problem, methods for measuring biomass and its components and cytokinesis.

Three yeast proteins that specifically inhibit yeast proteases A, B, and C.

Abstract: Baker9s yeast was found to contain inhibitors of yeast proteases A and C. These two proteins were partially purified, characterized, and compared with the previously described inhibitor of protease B. The A and B inhibitors were very thermostable and were extracted from intact yeast cells at 9k C. The A inhibitor appeared to be a protein with a molecular weight of about 22,000 which could be dissociated into two monomers or chains, both of which had a molecular weight of approximately 11,000. The protease C (carboxypeptidase Y)-inhibitor complex was purified and then partially disociated on an ion-exchange column. The free protease C inhibitor was very unstable, possibly because of destruction by a contaminating protease. Each inhibitor was specific for its corresponding protease and each inhibition was competitive. Whereas proteases A, B, and C destroyed the B inhibitor, only protease B had a pronounced destructive effect on the protease A inhibitor. Pepstatin was found to be a selective inhibitor of protease A, whereas chymostatin and antipain specifically inhibited protease B. Images

Molecular aspects of yeast morphogenesis.

Abstract: SEPTUM FORMATION 193 Structural Aspects. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 193 Molecular Aspects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195 Subcellular Organi.ation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . • . . . . . . . • . . . . . . 197 Compartmentation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 198 Septum Formation as a Model. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 199

Factors affecting filamentation in Candida albicans: changes in respiratory activity of Candida albicans during filamentation.

Abstract: Glucose metabolism and respiration of Candida albicans were compared under conditions which permitted either maximal filamentous or maximal yeast growth. Changes in metabolism were monitored by comparing the quantities of ethanol produced, CO2 evolved, and oxygen consumed. Filamenting cultures produced more ethanol and less CO2 than yeasts, with oxygen consumption in the former concomitantly slower than that of the latter. Studies involving cofactors and inhibitors associated with electron transport imply that a transfer of electrons away from flavoprotein is required for maintenance of yeast morphology. Conditions consistent with a buildup of reduced flavoprotein, however, favored filament formation. These changes were expressed metabolically as a shift from an aerobic to a fermentative metabolism. The results presented are consistent with hypotheses correlating filament production with changes in carbohydrate metabolism and an interruption of electron transfer within the cell.

Protein content and amino acid composition of certain fungi evaluated for microbial protein production.

Abstract: The protein and total amino acid contents of four mycelial fungal strains and one yeast were approximately the same for cultures harvested in the mid-log and early stationary growth phases. It was found that Fusarium oxysporum and Fusarium moniliforme contained approximately 30% more protein and total amino acids than Aspergillus niger. The amino acid composition of mycelial protein compares favorably with that of British Petroleum yeast protein Toprina produced commercially on hydrocarbon substrates. Fusarium spp. may be suitable for commercial production of microbial protein, especially when low-cost agricultural or industrial waste products are readily available as energy sources. Genetic manipulation of these fungi, such as induction of mutant strains through irradiation, may be desirable to obtain a mycelial product of improved yield and/or quality.

Translation of poly(riboadenylic acid)-enriched messenger RNAs from the yeast, Saccharomyces cerevisiae, in heterologous cell-free systems.

Abstract: Poly(riboadenylic acid) [poly(A)] enriched messenger RNAs from yeast have been used to direct the synthesis of yeast polypeptides in mouse Krebs II ascites and wheat embryo extracts. Both cell-free systems, synthesize polypeptides over a molecular weight range of 10,000-100,000. Autoradiograms of sodium dodecyl sulfate-polyacrylamide slab gels used to fractionate [35S]methionine-labeled in vitro products reveal that about 25 major bands (each of them possibly representing multiple polypeptides) are produced by each cell-free system. Each of these coelectrophoreses with a major polypeptide labeled in vivo or in a yeast lysate. These results suggest that cell-free translational machinery from eukaryotes is not able to discriminate in an all or none fashion against messenger RNAs which are available to it. While yeast poly(A)-enriched messenger RNA directs the synthesis polypeptides over approximately the same molecular weight range in both cell-free systems, the wheat germ system directs the incorporation of 45 times the amount of [3H]serine into Cl3CCOOH-precipitable polypeptides. This is in contrast to the 2.5-fold more efficient translation of hemoglobin mRNA in the wheat embryo extract. Thus, the extract from mammalian cells is able to translate mRNA from a lower plant with a much lower efficiency than it translates hemoglobin mRNA, and at a lower efficiency than is observed using a cell-free system from wheat embryos. This indicates that the wheat embryo system is the one of choice for translation of yeast messenger RNA.

Amino acid sequence of two functional sites in yeast glycogen phosphorylase.

Abstract: The structure of two functional sites in baker's yeast (Saccharomyces cerevisiae) glycogen phosphorylase (EC 2.4 1.1) was determined as part of a study on the evolution of regulatory enzymes. S-Carboxymethylated, MaBH4-reduced 32-P-labeled yeast phosphorylase a was cleaved with CNBr, thermolysin, and pepsin. Peptides labeled with 32-P or carrying the fluorescent pyridoxyl marker were isolated and purified using ion-exchange chromatography and gel filtration. CNBr cleavage yielded a single radioactive phosphopeptide (42 residues long) and one small fluorescent peptide with the unique sequence epsilon-Pxy-Lys-Phe-Val-Met. Thermolysin digestion gave rise to one radioactive octapeptide and two fluorescent peptides, 15 and 2 residues long, respectively. From a combination of substractive Edman degradations and digestion with yeast protease C, the sequence of the 32-P-labeled octapeptide was established. Phosphothreonine was identified as the sole phosphorylated amino acid, giving the following structure for the site involved in the covalent regulation of yeast phosphorylase: Leu-Thr(P) -Gly-Phe-Leu-Pro-Gln-Glu. The two fluorescent thermolytic peptides, together with two additional pyridoxyl peptides isolated after peptic digestion of the enzyme yielded the following sequence around the site binding pyridoxal-5'-P, the cofactor essential for phosphorylase activity: Ile-Ser-Thr-Ala-Gly-Thr-Glu-Ala-Ser-Gly-Thr-Ser-Asn-Met-Lys(P Pxy)-Phe-Val-Met. While the phosphorylated site bears no resemblance to the site of covalent control in vertebrate phosphorylases, the pyridoxal-P binding site in the yeast enayme displays remarkable homologies with its animal counterparts; the finding that 14 out of 18 amino acids are identical strongly suggests that the cofactor must be directly involved in catalysis.

Some considerations of the flocculation characteristics of ale and lager yeast strains

Abstract: While some ale yeast strains are able to flocculate when cultured in a defined medium of glucose, ammonium salts, vitamins and ions, others require the presence of a nitrogen-containing inducer in the growth medium. On the other hand, all flocculent lager strains examined to date are able to flocculate after being cultured in a defined medium and do not appear to require the addition of inducer material to the growth medium. The inducer material present in wort has been identified as peptide. By the use of ion exchange chromatography the peptide fraction that induces flocculation has been found to contain a high level of acidic amino acid residues with a very similar structure to that reported for the α-factor involved in sexual agglutination of haploid α and a cells of Sacch. cerevisiae. Studies on the adsorption of Ca++ ion by the cell wall failed to reveal any significant differences in total uptake between flocculent and non-flocculent cultures. It would appear that Ca++ ions are bound less tightly by non-flocculent cells than by flocculent cells. The contribution of calcium to flocculation is not the absolute amount of this ion adsorbed by the yeast cell wall but rather the stereo-specific manner by which it is bound, i.e., its position relative to the three-dimensional structure of the yeast cell wall.

Microbiology of Ripening Honey

Abstract: Two main groups of bacteria, classified as Gluconobacter and Lactobacillus, are present in ripening honey. A third bacterial group, classified as Zymomonas, and several types of yeast are occasionally isolated. Both in natural honey and in synthetic syrup the bacterial population decreases in the course of the ripening process. Lactobacillus and Gluconobacter disappear after minimum moisture (about 18%) is reached, but the former does so sooner than the latter. The presence of these bacteria in different parts of the bee has been also investigated.

Simple and sensitive procedure for screening yeast mutants that lyse at nonpermissive temperatures.

Abstract: After mutagenesis, surviving yeast cells are grown on plates at 25 C and later exposed to 37 C. The plates are then overlaid with a soft agar containing p-nitrophenylphosphate at pH 9.7. Lysed cells liberate alkaline phosphatase which gives rise to a yellow color on and around colonies.