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Saccharomyces cerevisiae  

PubMed Central

Yeast prions are infectious proteins that spread exclusively by mating. The frequency of prions in the wild therefore largely reflects the rate of spread by mating counterbalanced by prion growth slowing effects in the host. We recently showed that the frequency of outcross mating is about 1% of mitotic doublings with 23–46% of total matings being outcrosses. These findings imply that even the mildest forms of the [PSI+], [URE3] and [PIN+] prions impart > 1% growth/survival detriment on their hosts. Our estimate of outcrossing suggests that Saccharomyces cerevisiae is far more sexual than previously thought and would therefore be more responsive to the adaptive effects of natural selection compared with a strictly asexual yeast. Further, given its large effective population size, a growth/survival detriment of > 1% for yeast prions should strongly select against prion-infected strains in wild populations of Saccharomyces cerevisiae.

Kelly, Amy C.; Wickner, Reed B.



Saccharomyces cerevisiae exhibits a yAP-1-mediated adaptive response to malondialdehyde.  

PubMed Central

Malondialdehyde (MDA) is a highly reactive aldehyde generally formed as a consequence of lipid peroxidation. MDA has been inferred to have mutagenic and cytotoxic roles and possibly to be a participant in the onset of atherosclerosis. Wild-type Saccharomyces cerevisiae acquires resistance to a lethal dose (5 mM) of MDA following prior exposure to a nonlethal concentration (1 mM). This response was completely inhibited by cycloheximide (50 microg ml(-1)), indicating a requirement for protein synthesis for adaptation. Furthermore, we have examined the roles of glutathione (GSH), mitochondrial function, and yAP-1-mediated transcription in conferring resistance and adaptation to MDA. A yap1 disruption mutant exhibited the greatest sensitivity and was unable to adapt to MDA, implicating yAP-1 in both the adaptive response and constitutive survival. The effect of MDA on GSH mutants indicated a role for GSH in initial resistance, whereas resistance acquired through adaptation was independent of GSH. Likewise, respiratory mutants (petite mutants) were sensitive to MDA but were still able to mount an adaptive response similar to that of the wild type, excluding mitochondria from any role in adaptation. MDA was detected in yeast cells by the thiobarbituric acid test and subsequent high-pressure liquid chromatography separation. Elevated levels were detected following treatment with hydrogen peroxide. However, the MDA-adaptive response was independent of that to H2O2.

Turton, H E; Dawes, I W; Grant, C M



Saccharomyces cerevisiae Produces a Yeast Substance that Exhibits Estrogenic Activity in Mammalian Systems  

NASA Astrophysics Data System (ADS)

Partially purified lipid extracts of Saccharomyces cerevisiae contain a substance that displaces tritiated estradiol from rat uterine cytosol estrogen receptors. The yeast product induces estrogenic bioresponses in mammalian systems as measured by induction of progesterone receptors in cultured MCF-7 human breast cancer cells and by a uterotrophic response and progesterone receptor induction after administration to ovariectomized mice. The findings raise the possibility that bakers' yeast may be a source of environmental estrogens.

Feldman, David; Stathis, Peter A.; Hirst, Margaret A.; Price Stover, E.; Do, Yung S.; Kurz, Walter



Saccharomyces cerevisiae engineered for xylose metabolism ...  


Title: Saccharomyces cerevisiae engineered for xylose metabolism exhibits a ... XYL3, mRNA transcript levels for glycolytic, fermentative, and pentose phosphate ... ADH2, COX13, NDI1, and NDE1), and regulatory proteins (HAP4 and MTH1) ...


Engineering a Saccharomyces cerevisiae Wine Yeast That Exhibits Reduced Ethanol Production during Fermentation under Controlled Microoxygenation Conditions  

Microsoft Academic Search

We recently showed that expressing an H2O-NADH oxidase in Saccharomyces cerevisiae drastically reduces the intracellular NADH concentration and substantially alters the distribution of metabolic fluxes in the cell. Although the engineered strain produces a reduced amount of ethanol, a high level of acetaldehyde accumu- lates early in the process (1 g\\/liter), impairing growth and fermentation performance. To overcome these undesirable

Stephanie Heux; Jean-Marie Sablayrolles; Remy Cachon; Sylvie Dequin



Generation of a Novel Saccharomyces cerevisiae Strain That Exhibits Strong Maltose Utilization and Hyperosmotic Resistance Using Nonrecombinant Techniques  

PubMed Central

A yeast strain capable of leavening both unsugared and sweet bread dough efficiently would reduce the necessity of carrying out the expensive procedure of producing multiple baker's yeast strains. But issues involving the use of genetically modified foods have rendered the use of recombinant techniques for developing yeast strains controversial. Therefore, we used strong selection and screening systems in conjunction with traditional mass mating techniques to develop a strain of Saccharomyces cerevisiae that efficiently leavens both types of dough.

Higgins, Vincent J.; Bell, Philip J. L.; Dawes, Ian W.; Attfield, Paul V.



CTF4 (CHL15) mutants exhibit defective DNA metabolism in the yeast Saccharomyces cerevisiae.  

PubMed Central

We have analyzed the CTF4 (CHL15) gene, earlier identified in two screens for yeast mutants with increased rates of mitotic loss of chromosome III and artificial circular and linear chromosomes. Analysis of the segregation properties of circular minichromosomes and chromosome fragments indicated that sister chromatid loss (1:0 segregation) is the predominant mode of chromosome destabilization in ctf4 mutants, though nondisjunction events (2:0 segregation) also occur at an increased rate. Both inter- and intrachromosomal mitotic recombination levels are elevated in ctf4 mutants, whereas spontaneous mutation to canavanine resistance was not elevated. A genomic clone of CTF4 was isolated and used to map its physical and genetic positions on chromosome XVI. Nucleotide sequence analysis of CTF4 revealed a 2.8-kb open reading frame with a 105-kDa predicted protein sequence. The CTF4 DNA sequence is identical to that of POB1, characterized as a gene encoding a protein that associates in vitro with DNA polymerase alpha. At the N-terminal region of the protein sequence, zinc finger motifs which define potential DNA-binding domains were found. The C-terminal region of the predicted protein displayed similarity to sequences of regulatory proteins known as the helix-loop-helix proteins. Data on the effects of a frameshift mutation suggest that the helix-loop-helix domain is essential for CTF4 function. Analysis of sequences upstream of the CTF4 open reading frame revealed the presence of a hexamer element, ACGCGT, a sequence associated with many DNA metabolism genes in budding yeasts. Disruption of the coding sequence of CTF4 did not result in inviability, indicating that the CTF4 gene is nonessential for mitotic cell division. However, ctf4 mutants exhibit an accumulation of large budded cells with the nucleus in the neck. ctf4 rad52 double mutants grew very slowly and produced extremely high levels (50%) of inviable cell division products compared with either single mutant alone, which is consistent with a role for CTF4 in DNA metabolism. Images

Kouprina, N; Kroll, E; Bannikov, V; Bliskovsky, V; Gizatullin, R; Kirillov, A; Shestopalov, B; Zakharyev, V; Hieter, P; Spencer, F



Epigenetics in Saccharomyces cerevisiae.  


Saccharomyces cerevisiae provides a well-studied model system for heritable silent chromatin, in which a nonhistone protein complex--the SIR complex--represses genes by spreading in a sequence-independent manner, much like heterochromatin in higher eukaryotes. The ability to study mutations in histones and to screen genome-wide for mutations that impair silencing has yielded an unparalleled depth of detail about this system. Recent advances in the biochemistry and structural biology of the SIR-chromatin complex bring us much closer to a molecular understanding of how Sir3 selectively recognizes the deacetylated histone H4 tail and demethylated histone H3 core. The existence of appropriate mutants has also shown how components of the silencing machinery affect physiological processes beyond transcriptional repression. PMID:23818500

Grunstein, Michael; Gasser, Susan M



Peroxisome biogenesis in Saccharomyces cerevisiae  

Microsoft Academic Search

The observation that peroxisomes ofSaccharomyces cerevisiae can be induced by oleic acid has opened the possibility to investigate the biogenesis of these organelles in a biochemically and genetically well characterized organism. Only few enzymes have been identified as peroxisomal proteins inSaccharomyces cerevisiae so far; the three enzymes involved in ß-oxidation of fatty acids, enzymes of the glyoxylate cycle, catalase A

Wolf-H. Kunau; Andreas Hartig



Metabolite compartmentation in Saccharomyces cerevisiae.  

PubMed Central

Uninduced cultures of Saccharomyces cerevisiae exhibit high basal levels of allantoinase, allantoicase, and ureidoglycolate hydrolase, the enzymes responsible for degrading allantoin to urea. As a result, these activities increase only 4- to 8-fold upon induction, whereas the urea-degrading enzymes, urea carboxylase and allophanate hydrolase, have very low basal levels and routinely increase 30-fold on induction. Differences in the inducibility of these five enzymes were somewhat surprising because they are all part of the same pathway and have the same inducer, allophanate. Our current studies reconcile these observations. S. cerevisiae normally contained up to 1 mM allantoin sequestered in a cellular organelle, most likely the vacuole. Separation of the large amounts of allantoin and the enzymes that degrade it provide the cell with an efficient nitrogen reserve. On starvation, sequestered allantoin likely becomes accessible to these degradative enzymes. Because they are already present at high levels, the fact that their inducer is considerably removed from the input allantoin is of little consequence. This suggests that at times metabolite compartmentation may play an equal role with enzyme induction in the regulation of allantoin metabolism. Metabolism of arginine, another sequestered metabolite, must be controlled both by induction of arginase and compartmentation because arginine serves both as a reserve nitrogen source and a precursor of protein synthesis. The latter function precludes the existence of high basal levels of arginase.

Zacharski, C A; Cooper, T G



Xylose fermentation by Saccharomyces cerevisiae  

Microsoft Academic Search

We have performed a comparative study of xylose utilization in Saccharomyces cerevisiae transformants expressing two key enzymes in xylose metabolism, xylose reductase (XR) and xylitol dehydrogenase (XDH), and in a prototypic xylose-utilizing yeast, Pichia stipitis. In the absence of respiration (see text), baker's yeast cells convert half of the xylose to xylitol and ethanol, whereas P. stipilis cells display rather

Peter Kötter; Michael Ciriacy



Cadmium biosorption by Saccharomyces cerevisiae  

Microsoft Academic Search

Cadmium uptake by nonliving and resting cells of Saccharomyces cerevisiae obtained from aerobic or anaerobic cultures from pure cadmium-bearing solutions was examined. The highest cadmium uptake exceeding 70 mg Cd\\/g was observed with aerobic baker's yeast biomass from the exponential growth phase. Nearly linear sorption isotherms featured by higher sorbing resting cells together with metal deposits localized exclusively in vacuoles

B. Volesky; H. May; Z. R. Holan



Formation of cadaverine derivatives in Saccharomyces cerevisiae  

Microsoft Academic Search

The higher homologues of cadaverine, aminopropylcadaverine (APC) and N,N-bis(3-aminopropyl)cadaverine (3APC) were formed by a wild-type strain of Saccharomyces cerevisiae, and by two mutant strains, spe 3-1 and spe 4-1, exhibiting point mutations in the genes for spermidine synthase and spermine synthase, respectively. This, together with the incomplete inhibition of APC and 3 APC formation in the presence of inhibitors of

Dale R. Walters; Tracy Cowley



Fungal ?-glucosidase expression in Saccharomyces cerevisiae.  


Recombinant Saccharomyces cerevisiae strains expressing ?-glucosidases from Thermoascus aurantiacus (Tabgl1) and Phanerochaete chrysosporium (PcbglB and Pccbgl1) were constructed and compared to S. cerevisiae Y294[SFI], previously identified as the best ?-glucosidase-producing strain. The PcbglB was also intracellularly expressed in combination with the lac12 lactose permease of Kluyveromyces lactis in S. cerevisiae Y294[PcbglB + Lac12]. The recombinant extracellular ?-glucosidases indicated maximum activity in the pH range 4-5 and temperature optima varying from 50 to 75 °C. The S. cerevisiae Y294[Pccbgl1] strain performed best under aerobic and anaerobic conditions, producing 2.6 times more ?-glucosidase activity than S. cerevisiae Y294[SFI] and an ethanol concentration of 4.8 g l(-1) after 24 h of cultivation on cellobiose as sole carbohydrate source. S. cerevisiae Y294[Tabgl1] was unable to grow on cellobiose (liquid medium), whereas S. cerevisiae Y294[PcbglB + Lac12] exhibited limited growth. PMID:22707073

Njokweni, A P; Rose, S H; van Zyl, W H



Asparaginase II of Saccharomyces cerevisiae  

Microsoft Academic Search

The production of some extracellular enzymes is known to be negatively affected by readily metabolized nitrogen sources such\\u000a as NH4+ although there is no consensus regarding the involved mechanisms. Asparaginase II is a periplasmic enzyme ofSaccharomyces cerevisiae encoded by theASP3 gene. The enzyme activity is not found in cells grown in either ammonia, glutamine, or glutamate, but it is found

Elba P. S. Bon; Elvira Carvajal; Mike Stanbrough; Donald Rowen; Boris Magasanik



Apurinic endonucleases from Saccharomyces cerevisiae.  

PubMed Central

Three endonuclease activities have been partially purified from Saccharomyces cerevisiae on the basis of their activity against x-irradiated closed-circular supercoiled bacteriophage PM2 DNA. These endonucleases also nick apurinic DNA and two out of the three activities incise DNA UV-irradiated with high doses. The endonuclease activities have also been distinguished on the basis of their magnesium requirement and sensitivity to EDTA.

Armel, P R; Wallace, S S



Lead toxicity in Saccharomyces cerevisiae  

Microsoft Academic Search

The effect of Pb on Saccharomyces cerevisiae cell structure and function was examined. Membrane integrity was assessed by the release of UV-absorbing compounds and by\\u000a the intracellular K+ efflux. No leakage of UV260-absorbing compounds or loss of K+ were observed in Pb (until 1,000 ?mol\\/l) treated cells up to 30 min; these results suggest that plasma membrane seems not\\u000a to be the

Maarten Van der Heggen; Sara Martins; Gisela Flores; Eduardo V. Soares



Saccharomyces cerevisiae in directed evolution  

PubMed Central

Over the past 20 years, directed evolution has been seen to be the most reliable approach to protein engineering. Emulating the natural selection algorithm, ad hoc enzymes with novel features can be tailor-made for practical purposes through iterative rounds of random mutagenesis, DNA recombination and screening. Of the heterologous hosts used in laboratory evolution experiments, the budding yeast Saccharomyces cerevisiae has become the best choice to express eukaryotic proteins with improved properties. S. cerevisiae not only allows mutant enzymes to be secreted but also, it permits a wide range of genetic manipulations to be employed, ranging from in vivo cloning to the creation of greater molecular diversity, thanks to its efficient DNA recombination apparatus. Here, we summarize some successful examples of the use of the S. cerevisiae machinery to accelerate artificial evolution, complementing the traditional in vitro methods to generate tailor-made enzymes.

Gonzalez-Perez, David; Garcia-Ruiz, Eva; Alcalde, Miguel



Transgenic Saccharomyces Cerevisiae and Method for Bioremediation.  

National Technical Information Service (NTIS)

An isolated and purified transgenic Saccharomyces cerevisiae yeast cell comprising a disrupted ACR3 gene and an isolated DNA sequence comprising a promoter operably linked to a nucleic acid molecule encoding yeast cadmium factor resistance protein Ycf1p, ...

B. Rosen M. Ghosh



Nucleosome Positioning in Saccharomyces cerevisiae  

PubMed Central

Summary: The DNA of eukaryotic cells is spooled around large histone protein complexes, forming nucleosomes that make up the basis for a high-order packaging structure called chromatin. Compared to naked DNA, nucleosomal DNA is less accessible to regulatory proteins and regulatory processes. The exact positions of nucleosomes therefore influence several cellular processes, including gene expression, chromosome segregation, recombination, replication, and DNA repair. Here, we review recent technological advances enabling the genome-wide mapping of nucleosome positions in the model eukaryote Saccharomyces cerevisiae. We discuss the various parameters that determine nucleosome positioning in vivo, including cis factors like AT content, variable tandem repeats, and poly(dA:dT) tracts that function as chromatin barriers and trans factors such as chromatin remodeling complexes, transcription factors, histone-modifying enzymes, and RNA polymerases. In the last section, we review the biological role of chromatin in gene transcription, the evolution of gene regulation, and epigenetic phenomena.

Jansen, An; Verstrepen, Kevin J.



Choline transport in Saccharomyces cerevisiae.  

PubMed Central

Choline transport of Saccharomyces cerevisiae was measured by the filtration method with the use of glass microfiber paper. The uptake was time and temperature dependent. The kinetics of choline transport showed Michaelis behavior; an appearent Km for choline was 0.56 microM. N-Methylethanolamine, N,N-dimethylethanolamine, and beta-methylcholine were competitive inhibitors of choline transport, with Ki values of 40.1, 3.1, and 6.9 microM, respectively. Ethanolamine, phosphorylcholine, and various amino acids examined had no effect. Choline transport required metabolic energy; removal of glucose resulted in a great loss of transport activity, and the remaining activity was abolished by 2,4-dinitrophenol, carbonyl cyanide p-trifluoromethoxyphenyl hydrazone, arsenate, and cyanide. External Na+ was not required, and the transport was not effected by ionophores, valinomycin, and gramicidin D. These results indicate that S. cerevisiae possess an active choline transport system mediated by a specific carrier. This view is further supported by the isolation and characterization of a choline transport mutant. The choline transport activity in this mutant was very low, whereas the transport of L-leucine, L-methionine, D-glucose, and myo-inositol was normal. Together with the choline transport mutant, mutants defective in choline kinase were also isolated.

Hosaka, K; Yamashita, S



General and specific controls of lysine biosynthesis in Saccharomyces cerevisiae  

Microsoft Academic Search

Six of the eight enzymes of the a-aninoadipate pathway for the biosynthesis of lysine in Saccharomyces cerevisiae were examined for repressibility to lysine and for susceptibility to the general control of amino acid biosynthesis. All of the enzymes exhibited a 2 to 4 fold lower level of specific activity in the wildtype strain X2180 when grown in lysine supplemented medium

L. A. Urrestarazu; C. W. Borell; J. K. Bhattacharjee



The Maltose Permease Encoded by the Mal61 Gene of Saccharomyces Cerevisiae Exhibits Both Sequence and Structural Homology to Other Sugar Transporters  

PubMed Central

The MAL61 gene of Saccharomyces cerevisiae encodes maltose permease, a protein required for the transport of maltose across the plasma membrane. Here we report the nucleotide sequence of the cloned MAL61 gene. A single 1842 bp open reading frame is present within this region encoding the 614 residue putative MAL61 protein. Hydropathy analysis suggests that the secondary structure consists of two blocks of six transmembrane domains separated by an approximately 71 residue intracellular region. The N-terminal and C-terminal domains of 100 and 67 residues in length, respectively, also appear to be intracellular. Significant sequence and structural homology is seen between the MAL61 protein and the Saccharomyces high-affinity glucose transporter encoded by the SNF3 gene, the Kluyveromyces lactis lactose permease encoded by the LAC12 gene, the human HepG2 glucose transporter and the Escherichia coli xylose and arabinose transporters encoded by the xylE and araE genes, indicating that all are members of a family of sugar transporters and are related either functionally or evolutionarily. A mechanism for glucose-induced inactivation of maltose transport activity is discussed.

Cheng, Q.; Michels, C. A.



Physiolgical Roles of Pyruvate Decarboxylase in Saccharomyces Cerevisiae.  

National Technical Information Service (NTIS)

Contents: Pyruvate decarboxylase: in indispensable enzyme for growth of Saccharomyces cerevisiae on glucose; Growth requirements of pyruvate-decarboxylase-negative Saccharomyces cerevisiae strains; Metabolic responses of pyruvate-decarboxylase-negative Sa...

M. Filkweert



SCMD: Saccharomyces cerevisiae Morphological Database  

PubMed Central

To study the global regulation of cell morphology, a number of groups have recently reported genome-wide screening data for yeast mutants with abnormal morphology. Despite the relatively simple ellipsoidal shape of yeast cells, in the past, cell morphology researchers have processed information on cells manually. These time-consuming, entirely subjective tasks motivated us to develop image-processing software that automatically extracts yeast cells from micrographs and processes them to measure key morphological characteristics such as cell size, roundness, bud neck position angle, nuclear DNA localization and actin localization. To date, we have retrieved 960 609 cells from 52 988 micrographs of 2531 mutants using our software, and we have published the results in the Saccharomyces cerevisiae Morphological Database (SCMD), which facilitates the analysis of abnormal cells. Our system provides quantitative data for shapes of the daughter and mother cells, localization of the nuclear DNA and morphology of the actin patches. To search for mutants with similar morphological traits, the system outputs a list of mutants ranked by similarity of average morphological parameters. The SCMD is available at

Saito, Taro L.; Ohtani, Miwaka; Sawai, Hiroshi; Sano, Fumi; Saka, Ayaka; Watanabe, Daisuke; Yukawa, Masashi; Ohya, Yoshikazu; Morishita, Shinichi



Glucose Signaling in Saccharomyces cerevisiae  

PubMed Central

Eukaryotic cells possess an exquisitely interwoven and fine-tuned series of signal transduction mechanisms with which to sense and respond to the ubiquitous fermentable carbon source glucose. The budding yeast Saccharomyces cerevisiae has proven to be a fertile model system with which to identify glucose signaling factors, determine the relevant functional and physical interrelationships, and characterize the corresponding metabolic, transcriptomic, and proteomic readouts. The early events in glucose signaling appear to require both extracellular sensing by transmembrane proteins and intracellular sensing by G proteins. Intermediate steps involve cAMP-dependent stimulation of protein kinase A (PKA) as well as one or more redundant PKA-independent pathways. The final steps are mediated by a relatively small collection of transcriptional regulators that collaborate closely to maximize the cellular rates of energy generation and growth. Understanding the nuclear events in this process may necessitate the further elaboration of a new model for eukaryotic gene regulation, called “reverse recruitment.” An essential feature of this idea is that fine-structure mapping of nuclear architecture will be required to understand the reception of regulatory signals that emanate from the plasma membrane and cytoplasm. Completion of this task should result in a much improved understanding of eukaryotic growth, differentiation, and carcinogenesis.

Santangelo, George M.



Proteomics of Saccharomyces cerevisiae Organelles*  

PubMed Central

Knowledge of the subcellular localization of proteins is indispensable to understand their physiological roles. In the past decade, 18 studies have been performed to analyze the protein content of isolated organelles from Saccharomyces cerevisiae. Here, we integrate the data sets and compare them with other large scale studies on protein localization and abundance. We evaluate the completeness and reliability of the organelle proteomics studies. Reliability depends on the purity of the organelle preparations, which unavoidably contain (small) amounts of contaminants from different locations. Quantitative proteomics methods can be used to distinguish between true organellar constituents and contaminants. Completeness is compromised when loosely or dynamically associated proteins are lost during organelle preparation and also depends on the sensitivity of the analytical methods for protein detection. There is a clear trend in the data from the 18 organelle proteomics studies showing that proteins of low abundance frequently escape detection. Proteins with unknown function or cellular abundance are also infrequently detected, indicating that these proteins may not be expressed under the conditions used. We discuss that the yeast organelle proteomics studies provide powerful lead data for further detailed studies and that methodological advances in organelle preparation and in protein detection may help to improve the completeness and reliability of the data.

Wiederhold, Elena; Veenhoff, Liesbeth M.; Poolman, Bert; Slotboom, Dirk Jan



Dissection of Saccharomyces cerevisiae asci.  


Yeast is a highly tractable model system that is used to study many different cellular processes. The common laboratory strain Saccharomyces cerevisiae exists in either a haploid or diploid state. The ability to combine alleles from two haploids and the ability to introduce modifications to the genome requires the production and dissection of asci. Asci production from haploid cells begins with the mating of two yeast haploid strains with compatible mating types to produce a diploid strain. This can be accomplished in a number of ways either on solid medium or in liquid. It is advantageous to select for the diploids in medium that selectively promotes their growth compared to either of the haploid strains. The diploids are then allowed to sporulate on nutrient-poor medium to form asci, a bundle of four haploid daughter cells resulting from meiotic reproduction of the diploid. A mixture of vegetative cells and asci is then treated with the enzyme zymolyase to digest away the membrane sac surrounding the ascospores of the asci. Using micromanipulation with a microneedle under a dissection microscope one can pick up individual asci and separate and relocate the four ascopores. Dissected asci are grown for several days and tested for the markers or alleles of interest by replica plating onto appropriate selective media. PMID:19455096

Morin, Audrey; Moores, Adrian W; Sacher, Michael



Saccharomyces cerevisiae as a vaccine against coccidioidomycosis  

Microsoft Academic Search

Disseminated coccidioidomycosis is a life-threatening infection. In these studies, we examined protection against systemic murine coccidioidomycosis by vaccination with heat-killed Saccharomyces cerevisiae (HKY). CD-1 mice received HKY subcutaneously or by oral gavage with or without adjuvants once weekly beginning 3 or 4 weeks prior to infection; oral live Saccharomyces was also studied. All HKY sc regimens were equivalent, prolonging survival

Javier Capilla; Karl V. Clemons; Min Liu; Hillel B. Levine; David A. Stevens



40 CFR 180.1246 - Yeast Extract Hydrolysate from Saccharomyces cerevisiae: exemption from the requirement of a...  

Code of Federal Regulations, 2010 CFR

...Extract Hydrolysate from Saccharomyces cerevisiae: exemption from the requirement...Extract Hydrolysate from Saccharomyces cerevisiae: exemption from the...Extract Hydrolysate from Saccharomyces cerevisiae on all food...



40 CFR 180.1246 - Yeast Extract Hydrolysate from Saccharomyces cerevisiae: exemption from the requirement of a...  

Code of Federal Regulations, 2010 CFR

...Extract Hydrolysate from Saccharomyces cerevisiae: exemption from the requirement...Extract Hydrolysate from Saccharomyces cerevisiae: exemption from the...Extract Hydrolysate from Saccharomyces cerevisiae on all food...



Fatal Saccharomyces cerevisiae aortic graft infection.  


Saccharomyces cerevisiae is a yeast commonly used in baking and a frequent colonizer of human mucosal surfaces. It is considered relatively nonpathogenic in immunocompetent adults (J. N. Aucott, J. Fayan, H. Grossnicklas, A. Morrissey, M. M. Lederman, and R. A. Salata, Rev. Infect. Dis. 12:406-411, 1990). We present a case of S. cerevisiae fungemia and aortic graft infection in an immunocompetent adult. This is the first reported case of S. cerevisiae fungemia where the identity of the pathogen was confirmed by rRNA sequencing. PMID:12089311

Smith, Davey; Metzgar, David; Wills, Christopher; Fierer, Joshua



Genetic Characterization of Pathogenic Saccharomyces Cerevisiae Isolates  

PubMed Central

Saccharomyces cerevisiae isolates from human patients have been genetically analyzed. Some of the characteristics of these isolates are very different from laboratory and industrial strains of S. cerevisiae and, for this reason, stringent genetic tests have been used to confirm their identity as S. cerevisiae. Most of these clinical isolates are able to grow at 42°, a temperature that completely inhibits the growth of most other S. cerevisiae strains. This property can be considered a virulence trait and may help explain the presence of these isolates in human hosts. The ability to grow at 42° is shown to be polygenic with primarily additive effects between loci. S. cerevisiae will be a useful model for the evolution and genetic analysis of fungal virulence and the study of polygenic traits.

McCusker, J. H.; Clemons, K. V.; Stevens, D. A.; Davis, R. W.



Boron Stimulates Yeast (Saccharomyces cerevisiae) Growth  

Microsoft Academic Search

Boron is required for the growth of vascular plants and embryonic development in fish. The molecular basis of boron's essentiality, however, remains unknown for both. The objective of this study was to determine whether yeast (Saccharomyces cerevisiae) could be used as a model for the evaluation of intracellular boron traffick- ing. Three experiments were conducted to assess the ef- fect

A. Bennett; R. I. Rowe; N. Soch; C. D. Eckhert


Tangential Ultrafiltration of Aqueous "Saccharomyces Cerevisiae" Suspensions  

ERIC Educational Resources Information Center

|Experimental work on ultrafiltration is presented to illustrate the practical and theoretical principles of this separation technique. The laboratory exercise comprises experiments with pure water and with aqueous "Saccharomyces cerevisiae" (from commercial Baker's yeast) suspensions. With this work students detect the characteristic phenomena…

Silva, Carlos M.; Neves, Patricia S.; Da Silva, Francisco A.; Xavier, Ana M. R. B.; Eusebio, M. F. J.



Regulation of Biotin Transport in Saccharomyces Cerevisiae.  

National Technical Information Service (NTIS)

The metabolic control of biotin transport in Saccharomyces cerevisiae was investigated. Nonproliferating cells harvested from cultures grown in excess biotin (25 ng/ml) took up small amounts of biotin, whereas cells grown in biotin-sufficient medium (0.25...

T. O. Rogers H. C. Lichstein



Mechanism of Uranium mineralization by Saccharomyces cerevisiae  

NASA Astrophysics Data System (ADS)

We examined mechanism of the uranium mineralization by the yeast (Saccharomyces cerevisiae, X-2180-1B). Uptake experiments of U by S. cerevisiae were carried out for 4.2x10-4 M U solutions with their final pHs between 3 and 5. Concentrations of U and P were measured at the predetermined intervals. After 96 hours of contact time, the U accumulated S. cerevisiae was observed by scanning electron microscope equipped with energy dispersive spectroscopy (SEM-EDS). The Kd, defined by the ratio of U accumulated in S. cerevisiae to that in solution, shows monotone increase with time below pH 4, and reached steady state within 60 hours above pH 4. During U accumulation P was released from S. cerevisiae, while no release of P was observed in the absence of U. SEM-EDS analyses of the U accumulated S. cerevisiae indicated that uranyl phosphate minerals were occurred on the surface of S. cerevisiae at any pH. Thermodynamic calculation indicated that chemical compositions of the solutions at pH above 4 were super-saturated with respect to H-autunite. On the contrary, the chemical compositions of the solutions of pHs below 4 were under-saturated with respect to H-autunite, and were super-saturated at one-order higher concentration of U(VI) or P than observed. These results indicate that U mineralization is metabolism dependent, induced by the release of P from S. cerevisiae. Two different processes are relevant to U mineralization by S. cerevisiae. One is the process in which U reacts in the solution with P released from S. cerevisiae, where the chemical composition in the solution is super-saturated with respect to H-autunite. The other is the process in which U reacts with P on the surface of S. cerevisiae, where H-autunite is occurred by local-saturation condition.

Ohnuki, T.; Ozaki, T.; Yoshida, T.; Kozai, N.; Francis, A. J.; Iefuji, H.



Local Nanomechanical Motion of the Cell Wall of Saccharomyces cerevisiae  

Microsoft Academic Search

We demonstrate that the cell wall of living Saccharomyces cerevisiae (baker's yeast) exhibits local temperature-dependent nanomechanical motion at characteristic frequencies. The periodic motions in the range of 0.8 to 1.6 kHz with amplitudes of ~3 nm were measured using the cantilever of an atomic force microscope (AFM). Exposure of the cells to a metabolic inhibitor causes the periodic motion to

Andrew E. Pelling; Sadaf Sehati; Edith B. Gralla; Joan S. Valentine; James K. Gimzewski



Local Nanomechanical Motion of the Cell Wall of Saccharomyces cerevisiae  

Microsoft Academic Search

We demonstrate that the cell wall of living Saccharomyces cerevisiae (baker's yeast) exhibits local temperature-dependent nanomechanical motion at characteristic fre- quencies. The periodic motions in the range of 0.8 to 1.6 kHz with amplitudes of 3 nm were measured using the cantilever of an atomic force microscope (AFM). Exposure of the cells to a metabolic inhibitor causes the periodic motion

Andrew E. Pelling; Sadaf Sehati; Edith B. Gralla; Joan S. Valentine; James K. Gimzewski



Pseudouridine Mapping in the Saccharomyces cerevisiae Spliceosomal U Small Nuclear RNAs (snRNAs) Reveals that Pseudouridine Synthase Pus1p Exhibits a Dual Substrate Specificity for U2 snRNA and tRNA  

Microsoft Academic Search

Pseudouridine (C) residues were localized in the Saccharomyces cerevisiae spliceosomal U small nuclear RNAs (UsnRNAs) by using the chemical mapping method. In contrast to vertebrate UsnRNAs, S. cerevisiae UsnRNAs contain only a few C residues, which are located in segments involved in intermolecular RNA-RNA or RNA-protein interactions. At these positions, UsnRNAs are universally modified. When yeast mutants disrupted for one




l Asparaginase II of Saccharomyces cerevisiae  

Microsoft Academic Search

The activity profile of the periplasmic asparaginase of Saccharomyces cerevisiae was determined during cell growth in an ure2 mutant; in an ure2 transformed with a plasmid containing the gene URE2 and, for comparison, in the strain D273-10B. Cells were cultivated in media presenting variable quantitative and qualitative\\u000a nitrogen availability and the enzyme activity was evaluated in fresh and in nitrogen-starved

Edna M. M. Oliveira; Elvira Carvajal; Elba P. S. Bon



Saccharomyces cerevisiae aspartate kinase mechanism and inhibition  

Microsoft Academic Search

Aspartate kinase (AK) from Saccharomyces cerevisiae (AKsc) catalyzes the first step in the aspartate pathway responsible for biosynthesis of L-threonine, L-isoleucine, and L-methionine in fungi. Little was known about amino acids important for AKsc substrate binding and catalysis. Hypotheses about important amino acids were tested using site directed mutagenesis to substitute these amino acids with others having different properties. Steady

David Christopher Bareich



"Malonate Uptake and Metabolism in Saccharomyces cerevisiae".  


Malonyl-CoA plays an important role in the synthesis and elongation of fatty acids in yeast Saccharomyces cerevisiae. Malonyl-CoA is at a low concentration inside the cell and is produced mainly from acetyl-CoA through the enzyme acetyl-CoA carboxylase. It would be beneficial to find an alternative source of malonyl-CoA to increase its intracellular concentration and overall synthesis of the fatty acids. MatB gene from the bacteria Rhizobium leguminosarium bv. trifolii encodes for a malonyl-CoA synthetase which catalyzes the formation of the malonyl-CoA directly from malonate and CoA. However, results from high-performance liquid chromatography (HPLC) proved that Saccharomyces cerevisiae itself does not contain enough cytoplasmic malonate within them and is unable to uptake exogenously supplied malonate in the form of malonic acid. A dicarboxylic acid plasma membrane transporter with the ability to uptake exogenous malonic acid was identified from another species of yeast known as Schizosaccharomyces pombe and the gene encoding this transporter is identified as the mae1 gene. From the experiments thus far, the mae1 gene had been successfully cloned and transformed into Saccharomyces cerevisiae. The expression and functional ability of the encoded plasma membrane dicarboxylic acid transporter were also demonstrated and verified using specialized technologies such as RT-PCR, yeast immunofluorescence, HPLC, and LC-MS. PMID:23813405

Chen, Wei Ning; Tan, Kee Yang



RPL29 codes for a non-essential protein of the 60S ribosomal subunit in Saccharomyces cerevisiae and exhibits synthetic lethality with mutations in genes for proteins required for subunit coupling.  


RPL29 (YFR032c-a) is a non-essential gene that codes for a 60S ribosomal subunit protein in Saccharomyces cerevisiae. Deletion of RPL29 leads to a moderate accumulation of half-mer polysomes with little or no change in the amounts of free 60S subunits. In vitro translation and the growth rate are also delayed in the Deltarpl29 strain. Such a phenotype is characteristic of mutants defective in 60S to 40S subunit joining. The Deltarpl29 strain exhibits synthetic lethality with mutations in RPL10, the gene encoding an essential 60S ribosomal subunit protein that is required for 60S to 40S subunit joining. The Deltarpl29 strain also exhibits synthetic lethality with RSA1, a gene encoding a nucleoplasmic protein required for the loading of Rpl10p onto the 60S subunit. Over-expression of RPL10 suppresses the half-mer phenotype of the Deltarpl29 strain, but does not correct the growth defect of the deletion strain. We conclude that absence of Rpl29p impairs proper assembly of proteins onto the 60S subunit and that this retards subunit joining and additionally retards protein synthesis subsequent to subunit joining. PMID:11997090

DeLabre, Marie Laure; Kessl, Jacques; Karamanou, Spyridoula; Trumpower, Bernard L



Apoptosis - Triggering Effects: UVB-irradiation and Saccharomyces cerevisiae  

PubMed Central

ABSTRACT Objectives: The pathogenic disturbance of Saccharomyces cerevisiae is known as a rare but invasive nosocomial fungal infection. This survey is focused on the evaluation of apoptosis-triggering effects of UVB-irradiation in Saccharomyces cerevisiae. Materials and Methods: The well-growth colonies of Saccharomyces cerevisiae on Sabouraud Dextrose Agar (SDA) were irradiated within an interval of 10 minutes by UVB-light (302 nm). Subsequently, the harvested DNA molecules of control and UV-exposed yeast colonies were run through the 1% agarose gel electrophoresis comprising the luminescent dye of ethidium bromide. Outcomes: No unusual patterns including DNA laddering bands or smears were detected. Conclusions: The applied procedure for UV exposure was not effective for inducing apoptosis in Saccharomyces cerevisiae. So, it needs another UV-radiation protocol for inducing apoptosis phenomenon in Saccharomyces cerevisiae.




Cell Wall Assembly in Saccharomyces cerevisiae  

PubMed Central

An extracellular matrix composed of a layered meshwork of ?-glucans, chitin, and mannoproteins encapsulates cells of the yeast Saccharomyces cerevisiae. This organelle determines cellular morphology and plays a critical role in maintaining cell integrity during cell growth and division, under stress conditions, upon cell fusion in mating, and in the durable ascospore cell wall. Here we assess recent progress in understanding the molecular biology and biochemistry of cell wall synthesis and its remodeling in S. cerevisiae. We then review the regulatory dynamics of cell wall assembly, an area where functional genomics offers new insights into the integration of cell wall growth and morphogenesis with a polarized secretory system that is under cell cycle and cell type program controls.

Lesage, Guillaume; Bussey, Howard



Sporulation in the budding yeast Saccharomyces cerevisiae.  


In response to nitrogen starvation in the presence of a poor carbon source, diploid cells of the yeast Saccharomyces cerevisiae undergo meiosis and package the haploid nuclei produced in meiosis into spores. The formation of spores requires an unusual cell division event in which daughter cells are formed within the cytoplasm of the mother cell. This process involves the de novo generation of two different cellular structures: novel membrane compartments within the cell cytoplasm that give rise to the spore plasma membrane and an extensive spore wall that protects the spore from environmental insults. This article summarizes what is known about the molecular mechanisms controlling spore assembly with particular attention to how constitutive cellular functions are modified to create novel behaviors during this developmental process. Key regulatory points on the sporulation pathway are also discussed as well as the possible role of sporulation in the natural ecology of S. cerevisiae. PMID:22084423

Neiman, Aaron M



Protein arginine methylation in Saccharomyces cerevisiae.  


Recent research has implicated arginine methylation as a major regulator of cellular processes, including transcription, translation, nucleocytoplasmic transport, signalling, DNA repair, RNA processing and splicing. Arginine methylation is evolutionarily conserved, and it is now thought that it may rival other post-translational modifications such as phosphorylation in terms of its occurrence in the proteome. In addition, multiple recent examples demonstrate an exciting new theme: the interplay between methylation and other post-translational modifications such as phosphorylation. In this review, we summarize our current understanding of arginine methylation and the recent advances made, with a focus on the lower eukaryote Saccharomyces cerevisiae. We cover the types of methylated proteins, their responsible methyltransferases, where and how the effects of arginine methylation are seen in the cell, and, finally, discuss the conservation of the biological function of methylarginines between S. cerevisiae and mammals. PMID:23094907

Low, Jason K K; Wilkins, Marc R



Assessing chronological aging in Saccharomyces cerevisiae.  


Saccharomyces cerevisiae is one of the most studied model organisms for the identification of genes and mechanisms that affect aging. The chronological lifespan (CLS) assay, which monitors the survival of a non-dividing population, is one of the two methods to study aging in yeast. To eliminate potential artifacts and identify genes and signaling pathways that may also affect aging in higher eukaryotes, it is important to determine CLS by multiple methods. Here, we describe these methods as well as the assays to study macromolecular damage during aging in yeast, with a focus on genomic instability. PMID:23296677

Hu, Jia; Wei, Min; Mirisola, Mario G; Longo, Valter D



[Vaginal infection caused by Saccharomyces cerevisiae].  


In a woman of 26, who suffered from a vulvovaginal infection and had previously been treated for Candida vaginitis, Saccharomyces cerevisiae was cultured and identified. At her work she sold baking yeast. Topical treatment with amphotericin B 100 mg suppositories was successful. Microscopic examination (1000 x) of the discharge in saline showed haloed yeast cells. For treatment, oral ketoconazole or topical administration of amphotericin B or clotrimazole, in relatively high doses, may be applied. This yeast might be the cause of 'chronic candidiasis' more often than suspected, notably in women working in a bakery or a brewery. PMID:7783803

van Doorn, H C; Coelingh Bennink, F



Evidence for Domesticated and Wild Populations of Saccharomyces cerevisiae  

Microsoft Academic Search

Saccharomyces cerevisiae is predominantly found in association with human activities, particularly the production of alcoholic beverages. S. paradoxus, the closest known relative of S. cerevisiae, is commonly found on exudates and bark of deciduous trees and in associated soils. This has lead to the idea that S. cerevisiae is a domesticated species, specialized for the fermentation of alcoholic beverages, and

Justin C. Fay; Joseph A. Benavides



Specificities of the Saccharomyces Cerevisiae Rad6, Rad18, and Rad52 Mutators Exhibit Different Degrees of Dependence on the Rev3 Gene Product, a Putative Nonessential DNA Polymerase  

PubMed Central

The Saccharomyces cerevisiae rad6, rad18, and rad52 mutants exhibit DNA repair deficiencies and distinct mutator phenotypes. DNA replication past unrepaired spontaneous damage might contribute to the specificities of these mutators. Because REV3 is thought to encode a DNA polymerase that specializes in translesion synthesis, we determined the REV3 dependence of the rad mutator specificities. Spontaneous mutagenesis at a plasmid-borne SUP4-o locus was examined in isogenic strains having combinations of normal or mutant REV3 and RAD6, RAD18, or RAD52 alleles. For the rad6 and rad18 mutators, the mutation rate increase relied largely, but not exclusively, on REV3 whereas the rad52 mutator was entirely REV3 dependent. The influence of REV3 on the specificity of the rad6 mutator differed markedly depending on the mutational class examined. However, the requirement of rev3 for the production of G·C -> T·A transversions by the rad18 mutator, which induces only these substitutions, was similar to that for rad6-mediated G·C -> T·A transversion. This supports a role for the Rad6-Rad18 protein complex in the control of spontaneous mutagenesis. The available data imply that the putative Rev3 polymerase can process a variety of spontaneous DNA lesions that normally are substrates for error-free repair.

Roche, H.; Gietz, R. D.; Kunz, B. A.



Myo-inositol transport in Saccharomyces cerevisiae.  


myo-Inositol uptake in Saccharomyces cerevisiae was dependent on temperature, time, and substrate concentration. The transport obeyed saturation kinetics with an apparent Km for myo-inositol of 0.1 mM, myo-Inositol analogs, such as scyllo-inositol, 2-inosose, mannitol, and 1,2-cyclohexanediol, had no effect on myo-inositol uptake, myo-Inositol uptake required metabolic energy. Removal of D-glucose resulted in a loss of activity, and azide and cyanide ions were inhibitory. In the presence of D-glucose, myo-inositol was accumulated in the cells against a concentration gradient. A myo-inositol transport mutant was isolated from UV-mutagenized S. cerevisiae cells using the replica-printing technique. The defect in myo-inositol uptake was due to a single nuclear gene mutation. The activities of L-serine and D-glucose transport were not affected by the mutation. Thus it was shown that S. cerevisiae grown under the present culture conditions possessed a single and specific myo-inositol transport system. myo-Inositol transport activity was reduced by the addition of myo-inositol to the culture medium. The activity was reversibly restored by the removal of myo-inositol from the medium. This restoration of activity was completely abolished by cycloheximide. PMID:7040334

Nikawa, J; Nagumo, T; Yamashita, S



Expression of VHHs in Saccharomyces cerevisiae.  


The production of VHHs in microorganisms is relatively straightforward, however the amount of VHH produced per volume unit can vary substantially from hardly detectable to hundreds of milligrams per liter. Expression in Escherichia coli is more commonly used at initial research phase, since production of VHHs for large-scale application in E. coli is for a number of reasons not preferred. Otherwise VHH production in GRAS organisms such as Saccharomyces cerevisiae fits very well with industrial fermentation processes, and in fact the only commercially available VHHs are produced in S. cerevisiae. Immediately after the discovery of heavy chain only antibodies, which are per definition devoid of light chains, it was investigated whether many problems encountered with the production of conventional antibodies in lower eukaryotes were absent during the production of VHHs. Here we provide a protocol for the expression of VHH genes in S. cerevisiae in a fed-batch fermentation process. This protocol is also suitable for the production of multivalent VHHs. PMID:22886258

Gorlani, Andrea; de Haard, Hans; Verrips, Theo



A novel class of Saccharomyces cerevisiae mutants specifically UV-sensitive to “petite” induction  

Microsoft Academic Search

A mutant of Saccharomyces cerevisiae has been isolated which, though exhibiting a normal response to nuclear genetic damage by ultraviolet light (UV), is more sensitive than its wild type specifically in the production of the cytoplasmic (rho-) mutation by this agent. Some of the features of this mutation which has been designated uvs?5 are:i)The mutation is recessive, it exhibits a

Ethel Moustacchi; Philip S. Perlman; Henry R. Mahler



Genotypic and Physiological Characterization of Saccharomyces boulardii, the Probiotic Strain of Saccharomyces cerevisiae  

Microsoft Academic Search

Saccharomyces boulardii, a yeast that was isolated from fruit in Indochina, has been used as a remedy for diarrhea since 1950 and is now a commercially available treatment throughout Europe, Africa, and South America. Though initially classified as a separate species of Saccharomyces, recent publications have shown that the genome of S. boulardii is so similar to Saccharomyces cerevisiae that

Laura Edwards-Ingram; Paul Gitsham; Nicola Burton; Geoff Warhurst; Ian Clarke; David Hoyle; Stephen G. Oliver; Lubomira Stateva



Induction of respiration deficient mutants in Saccharomyces cerevisiae by Berenil  

Microsoft Academic Search

Some characteristic details of mutagenesis by Berenil, a non-intercalating trypanocidal dye, that govern the change from wild type (?+) to vegetative petite (?-) in Saccharomyces cerevisiae are presented and contrasted with the intercalating mutagens ethidium bromide and euflavine.

Philip S. Perlman; Henry R. Mahler



Effect of Sterol Alterations on Conjugation in Saccharomyces Cerevisiae.  

National Technical Information Service (NTIS)

Sterol auxotrophic strains of Saccharomyces cerevisiae were grown and allowed to conjugate on media supplemented with various sterols. The mating efficiency of the auxotrophs is perturbed by the replacement of the normal yeast sterol, ergosterol. with oth...

M. E. Tomeo G. Fenner S. R. Tove L. W. Parks



Septin filament organization in Saccharomyces cerevisiae  

PubMed Central

Septins are a family of GTP-binding, membrane-interacting cytoskeletal proteins, highly conserved and essential in all eukaryotes (with the exception of plants). Septins play important roles in a number of cellular events that involve membrane remodeling and compartmentalization. One such event is cytokinesis, the last stage of cell division. While cytokinesis is ultimately achieved via the mechanical contraction of an actomyosin ring at the septum, determination of the location where cytokinesis will take place, and recruitment of factors involved in signaling events leading to septation requires the activity of septins. We are working towards dissecting the properties of septins from the budding yeast Saccharomyces cerevisiae, where they were first discovered as cell cycle mutants. In our studies we have employed several complementary electron microscopy techniques to describe the organization and structure of septins both in vitro and in situ.

Bertin, Aurelie; Nogales, Eva



Viruses and prions of Saccharomyces cerevisiae.  


Saccharomyces cerevisiae has been a key experimental organism for the study of infectious diseases, including dsRNA viruses, ssRNA viruses, and prions. Studies of the mechanisms of virus and prion replication, virus structure, and structure of the amyloid filaments that are the basis of yeast prions have been at the forefront of such studies in these classes of infectious entities. Yeast has been particularly useful in defining the interactions of the infectious elements with cellular components: chromosomally encoded proteins necessary for blocking the propagation of the viruses and prions, and proteins involved in the expression of viral components. Here, we emphasize the L-A dsRNA virus and its killer-toxin-encoding satellites, the 20S and 23S ssRNA naked viruses, and the several infectious proteins (prions) of yeast. PMID:23498901

Wickner, Reed B; Fujimura, Tsutomu; Esteban, Rosa



Killer systems of the yeast Saccharomyces cerevisiae  

SciTech Connect

The killer systems of Saccharomyces cerevisiae are an unusual class of cytoplasmic symbionts of primitive eukaryotes. The genetic material of these symbionts is double-stranded RNA. They are characterized by the linearity of the genome, its fragmentation into a major and a minor fraction, which replicate separately, and their ability to control the synthesis of secretory mycocin proteins possessing a toxic action on closely related strains. The secretion of mycocins at the same time ensures acquiring of resistance to them. Strains containing killer symbionts are toxigenic and resistant to the action of their own toxin, but strains that are free of killer double-stranded RNAs are sensitive to the action of mycocins. The killer systems of S. cerevisiae have retained features relating them to viruses and are apparently the result of evolution of infectious viruses. The occurrences of such systems among monocellular eukaryotic organisms is an example of complication of the genome by means of its assembly from virus-like components. We discuss the unusual features of replication and the expression of killer systems and their utilization in the construction of vector molecules.

Nesterova, G.F.




PubMed Central

DPP1-encoded and LPP1-encoded lipid phosphate phosphatases are integral membrane proteins in the yeast Saccharomyces cerevisiae. They catalyze the Mg2+-independent dephosphorylation of bioactive lipid phosphate molecules such as diacylglycerol pyrophosphate and phosphatidate. These enzymes possess a three-domain lipid phosphatase motif that is localized to the hydrophilic surface of the membrane. The lipid phosphate phosphatase activities of DPP1-encoded and LPP1-encoded enzymes are measured by following the release of water-soluble radioactive inorganic phosphate from chloroform-soluble radioactive lipid phosphate substrate following a chloroform/methanol/water phase partition. The DPP1-encoded enzyme, commonly referred to as diacylglycerol pyrophosphate phosphatase, is purified from wild-type S. cerevisiae membranes by detergent solubilization with Triton X-100 followed by chromatography with DEAE-cellulose (DE53), Affi-Gel blue, hydroxylapatite, and Mono Q. The purification scheme yields an essentially homogeneous enzyme preparation that is stable for several years upon storage at ?80°. The properties of the DPP1-encoded and LPP1-encoded lipid phosphate phosphatase enzymes are summarized.

Carman, George M.; Wu, Wen-I



Molecular characterization of Saccharomyces cerevisiae TFIID.  


We previously defined Saccharomyces cerevisiae TFIID as a 15-subunit complex comprised of the TATA binding protein (TBP) and 14 distinct TBP-associated factors (TAFs). In this report we give a detailed biochemical characterization of this general transcription factor. We have shown that yeast TFIID efficiently mediates both basal and activator-dependent transcription in vitro and displays TATA box binding activity that is functionally distinct from that of TBP. Analyses of the stoichiometry of TFIID subunits indicated that several TAFs are present at more than 1 copy per TFIID complex. This conclusion was further supported by coimmunoprecipitation experiments with a systematic family of (pseudo)diploid yeast strains that expressed epitope-tagged and untagged alleles of the genes encoding TFIID subunits. Based on these data, we calculated a native molecular mass for monomeric TFIID. Purified TFIID behaved in a fashion consistent with this calculated molecular mass in both gel filtration and rate-zonal sedimentation experiments. Quite surprisingly, although the TAF subunits of TFIID cofractionated as a single complex, TBP did not comigrate with the TAFs during either gel filtration chromatography or rate-zonal sedimentation, suggesting that TBP has the ability to dynamically associate with the TFIID TAFs. The results of direct biochemical exchange experiments confirmed this hypothesis. Together, our results represent a concise molecular characterization of the general transcription factor TFIID from S. cerevisiae. PMID:12138208

Sanders, Steven L; Garbett, Krassimira A; Weil, P Anthony



Thermal Denaturation Kinetics of Yeast Proteins in Whole Cells of Saccharomyces cerevisiae and Kluyveromyces fragilis  

Microsoft Academic Search

The kinetics of thermal denaturation of yeast proteins in intact cells of Saccharomyces cerevisiae instant dry yeast and Kluyveromyces fragilis L-1930 have been studied through differential scanning calorimetry. Maximum deflection-peak temperatures (TP) were greater for S. cerevisiae (66.65°C) than for K. fragilis (63.21 °C). Kinetic parameters showed that the former was slightly more resistant to thermal protein denaturation and exhibited

M. A. Otero; J. R. Wagner; M. C. Vasallo; M. C. Añón; L. García; J. C. Jiménez; J. C. López



Alcoholic fermentation of xylose and mixed sugars using recombinant Saccharomyces cerevisiae engineered for xylose utilization  

Microsoft Academic Search

Previously, a Saccharomyces cerevisiae strain was engineered for xylose assimilation by the constitutive overexpression of the Orpinomyces xylose isomerase, the S. cerevisiae xylulokinase, and the Pichia stipitis SUT1 sugar transporter genes. The recombinant strain exhibited growth on xylose, under aerobic conditions, with a specific growth\\u000a rate of 0.025 h?1, while ethanol production from xylose was achieved anaerobically. In the present study,

Anjali Madhavan; Sriappareddy Tamalampudi; Aradhana Srivastava; Hideki Fukuda; Virendra S. Bisaria; Akihiko Kondo



Regulation of xylose metabolism in recombinant Saccharomyces cerevisiae  

Microsoft Academic Search

BACKGROUND: Considerable interest in the bioconversion of lignocellulosic biomass into ethanol has led to metabolic engineering of Saccharomyces cerevisiae for fermentation of xylose. In the present study, the transcriptome and proteome of recombinant, xylose-utilising S. cerevisiae grown in aerobic batch cultures on xylose were compared with those of glucose-grown cells both in glucose repressed and derepressed states. The aim was

Laura Salusjärvi; Matti Kankainen; Rabah Soliymani; Juha-Pekka Pitkänen; Merja Penttilä; Laura Ruohonen



Differential regulation of STA genes of Saccharomyces cerevisiae  

Microsoft Academic Search

The single glucoamylase gene (SGA1) of the yeast Saccharomyces cerevisiae is expressed exclusively during the sporulation phase of the life cycle. Enzymatic studies and nucleic acid sequence comparisons have shown that the SGA1 glucoamylase is closely related to the secreted enzymes of S. cerevisiae var. diastaticus. The latter are encoded by any of three unlinked STA genes, which have been

Tom A. Pugh; Mary J. Clancy



An assay for functional xylose transporters in Saccharomyces cerevisiae.  


It has been considered that more efficient uptake of xylose could promote increased xylose metabolic capacity of several microorganisms. In this study, an assay to screen xylose transporters was established in the Saccharomyces cerevisiae strain, which expresses the xylosidase gene of Bacillus pumilus intracellularly. The absorbed xylose analog p-nitrophenyl-?-d-xylopyranoside (pNPX) rapidly hydrolyzed to p-nitrophenol (pNP), which displayed a yellow tint when exposed to xylosidase in vivo. The xylose transporter activities of the strain were computed using the pNP production rate, which was detected extracellularly. This method could be used for both high-throughput screening and smaller scale investigations. AraEp, which is a pentose transporter of Corynebacterium glutamicum, was expressed in S. cerevisiae and exhibited better transport capacity than the endogenous transporters Hxt7p and Gal2p. Moreover, a mutant of AraEp with 103% greater transport capacity was screened out, and the computer simulation suggested that transmembrane domain 5 was an important factor for the transport capacity of AraEp in S. cerevisiae. PMID:23928049

Wang, Chengqiang; Shen, Yu; Hou, Jin; Suo, Fan; Bao, Xiaoming



Cybernetic model of the growth dynamics of Saccharomyces cerevisiae in batch and continuous cultures  

Microsoft Academic Search

Growth of Saccharomyces cerevisiae on glucose in aerobic batch culture follows the well-documented diauxic pattern of completely fermenting glucose to ethanol during the first exponential growth phase, followed by an intermediate lag phase and a second exponential growth phase consuming ethanol. In continuous cultures over a range of intermediate dilution rates, the yeast bioreactor exhibits sustained oscillations in all the

Kenneth D. Jones; Dhinakar S. Kompala



Induction and elimination of oscillations in continuous cultures of Saccharomyces cerevisiae  

Microsoft Academic Search

Continuous cultures of Saccharomyces cerevisiae are known to exhibit oscillatory behavior in the oxidative region. Important findings of a series of experiments conducted to identify the causes for initiation of and the means for elimination of oscillations in these cultures are reported in this paper. These oscillations are seen to be connected to the growth kinetics of the microorganism and

Satish J. Parulekar; Gary B. Semones; Michael J. Rolf; Jefferson C. Lievense; Henry C. Lim



Detection of anti- Saccharomyces cerevisiae antibodies in Crohn's disease: is it reliable diagnostic and prognostic marker?  

Microsoft Academic Search

Background. In the past few years, serologic markers have been proposed in inflammatory bowel disease. Anti-Saccharomyces cerevisiae antibodies showed high specificity for Crohn's disease. A prognostic role for serology has also been hypothesised.Aims To evaluate anti-Saccharomyces cerevisiae antibody distribution in an unselected Italian inflammatory bowel disease population. To analyse whether anti-Saccharomyces cerevisiae antibody status (positive\\/negative) and\\/or anti-Saccharomyces cerevisiae antibody titres

R. Sostegni; M. Daperno; E. Ercole; C. Rigazio; F. Bresso; G. Masoero; F. Castellino; C. Zaffino; R. Rocca; G. C. Molinaro; G. Rocca; M. Astegiano; A. Pera



The endopolyphosphatase gene: Essential in Saccharomyces cerevisiae  

PubMed Central

Endopolyphosphatases (Ppn1) from yeast and animal cells hydrolyze inorganic polyphosphate (poly P) chains of many hundreds of phosphate residues into shorter lengths. The limit digest consists predominantly of chains of 60 (P60) and 3 (P3) Pi residues. Ppn1 of Saccharomyces cerevisiae, a homodimer of 35-kDa subunits (about 352-aa) is of vacuolar origin and requires the protease activation of a 75-kDa (674-aa) precursor polypeptide. The Ppn1 gene (PPN1) now has been cloned, sequenced, overexpressed, and deleted. That PPN1 encodes Ppn1 was verified by a 25-fold increase in Ppn1 when overexpressed under a GAL promoter and also by several peptide sequences that match exactly with sequences in a yeast genome ORF, the mutation of which abolishes Ppn1 activity. Null mutants in Ppn1 accumulate long-chain poly P and are defective in growth in minimal media. A double mutant of PPN1 and PPX1 (the gene encoding a potent exopolyphosphatase) loses viability rapidly in stationary phase. Whether this loss is a result of the excess of long-chain poly P or to the lack of shorter chains (i.e., poly P60 and P3) is unknown. Overexpression of the processed form of Ppn1 should provide a unique and powerful reagent to analyze poly P when the chain termini are unavailable to the actions of polyPase and poly P kinase.

Sethuraman, Anand; Rao, Narayana N.; Kornberg, Arthur



Stationary phase in the yeast Saccharomyces cerevisiae.  

PubMed Central

Growth and proliferation of microorganisms such as the yeast Saccharomyces cerevisiae are controlled in part by the availability of nutrients. When proliferating yeast cells exhaust available nutrients, they enter a stationary phase characterized by cell cycle arrest and specific physiological, biochemical, and morphological changes. These changes include thickening of the cell wall, accumulation of reserve carbohydrates, and acquisition of thermotolerance. Recent characterization of mutant cells that are conditionally defective only for the resumption of proliferation from stationary phase provides evidence that stationary phase is a unique developmental state. Strains with mutations affecting entry into and survival during stationary phase have also been isolated, and the mutations have been shown to affect at least seven different cellular processes: (i) signal transduction, (ii) protein synthesis, (iii) protein N-terminal acetylation, (iv) protein turnover, (v) protein secretion, (vi) membrane biosynthesis, and (vii) cell polarity. The exact nature of the relationship between these processes and survival during stationary phase remains to be elucidated. We propose that cell cycle arrest coordinated with the ability to remain viable in the absence of additional nutrients provides a good operational definition of starvation-induced stationary phase.

Werner-Washburne, M; Braun, E; Johnston, G C; Singer, R A



Hypervariable Noncoding Sequences in Saccharomyces cerevisiae  

PubMed Central

Compared to protein-coding sequences, the evolution of noncoding sequences and the selective constraints placed on these sequences is not well characterized. To compare the evolution of coding and noncoding sequences, we have conducted a survey for DNA polymorphism at five randomly chosen loci among a diverse collection of 81 strains of Saccharomyces cerevisiae. Average rates of both polymorphism and divergence are 40% lower at noncoding sites and 90% lower at nonsynonymous sites in comparison to synonymous sites. Although noncoding and coding sequences show substantial variability in ratios of polymorphism to divergence, two of the loci, MLS1 and PDR10, show a higher rate of polymorphism at noncoding compared to synonymous sites. The high rate of polymorphism is not accompanied by a high rate of divergence and is limited to a few small regions. These hypervariable regions include sites with three segregating bases at a single site and adjacent polymorphic sites. We show that this clustering of polymorphic sites is significantly greater than one would expect on the basis of the spacing between polymorphic fourfold degenerate sites. Although hypervariable noncoding sequences could result from selection on regulatory mutations, they could also result from transient mutational hotspots.

Fay, Justin C.; Benavides, Joseph A.



Phosphate transport and sensing in Saccharomyces cerevisiae.  

PubMed Central

Cellular metabolism depends on the appropriate concentration of intracellular inorganic phosphate; however, little is known about how phosphate concentrations are sensed. The similarity of Pho84p, a high-affinity phosphate transporter in Saccharomyces cerevisiae, to the glucose sensors Snf3p and Rgt2p has led to the hypothesis that Pho84p is an inorganic phosphate sensor. Furthermore, pho84Delta strains have defects in phosphate signaling; they constitutively express PHO5, a phosphate starvation-inducible gene. We began these studies to determine the role of phosphate transporters in signaling phosphate starvation. Previous experiments demonstrated a defect in phosphate uptake in phosphate-starved pho84Delta cells; however, the pho84Delta strain expresses PHO5 constitutively when grown in phosphate-replete media. We determined that pho84Delta cells have a significant defect in phosphate uptake even when grown in high phosphate media. Overexpression of unrelated phosphate transporters or a glycerophosphoinositol transporter in the pho84Delta strain suppresses the PHO5 constitutive phenotype. These data suggest that PHO84 is not required for sensing phosphate. We further characterized putative phosphate transporters, identifying two new phosphate transporters, PHO90 and PHO91. A synthetic lethal phenotype was observed when five phosphate transporters were inactivated, and the contribution of each transporter to uptake in high phosphate conditions was determined. Finally, a PHO84-dependent compensation response was identified; the abundance of Pho84p at the plasma membrane increases in cells that are defective in other phosphate transporters.

Wykoff, D D; O'Shea, E K



Copper transport in the yeast Saccharomyces cerevisiae  

SciTech Connect

Biochemical processes involved in the movement of copper (Cu) into and out of the yeast Saccharomyces Cerevisiae have been investigated. Overall uptake of Cu was measured by disappearance of Cu from the reaction mixture by atomic absorption sensitive to 10/sup -10/M. The process of Cu influx is composed of a prerequisite binding and subsequent transport. The binding is non-energetic but is competitively inhibited by zinc(Zn). Transport is energetic as shown by an increased influx in the presence of added glucose. This process is prevented by 2,4-dinitrophenol(DNP). Cu influx is accompanied by an exchange for potassium(K) in a ratio of K:Cu=2:1. The process of Cu efflux involves a second type of binding site, probably of low affinity but large capacity. The presence of glucose causes the binding of extracellular Cu to these sites in a non-energy-dependent mechanism which prevents Cu efflux. Zn does not compete. DNP has no effect. The K:Cu ratio of 4:1 observed in the absence of glucose suggests a lowered net Cu uptake as a result of concomitant efflux activity. Finally, in the absence but not the presence of glucose, the pH of the extracellular solution increases. These observations are consistent with the idea that (a) yeast membrane has two Cu-binding sites, one of which participates in influx and one in efflux; (b) Cu exchanges with K during influx and with protons during efflux.

Martinez, L.D.; Connelly, J.L.



Local Nanomechanical Motion of the Cell Wall of Saccharomyces cerevisiae  

NASA Astrophysics Data System (ADS)

We demonstrate that the cell wall of living Saccharomyces cerevisiae (baker's yeast) exhibits local temperature-dependent nanomechanical motion at characteristic frequencies. The periodic motions in the range of 0.8 to 1.6 kHz with amplitudes of ~3 nm were measured using the cantilever of an atomic force microscope (AFM). Exposure of the cells to a metabolic inhibitor causes the periodic motion to cease. From the strong frequency dependence on temperature, we derive an activation energy of 58 kJ/mol, which is consistent with the cell's metabolism involving molecular motors such as kinesin, dynein, and myosin. The magnitude of the forces observed (~10 nN) suggests concerted nanomechanical activity is operative in the cell.

Pelling, Andrew E.; Sehati, Sadaf; Gralla, Edith B.; Valentine, Joan S.; Gimzewski, James K.



A Saccharomyces cerevisiae -based bioassay for assessing pesticide toxicity  

Microsoft Academic Search

This study evaluates the toxic effect of three pesticides (Azoxystrobin, Cymoxanil, and Diuron) on the yeast Saccharomyces cerevisiae for the development of a new bioassay based on inhibition of S. cerevisiae metabolic activity at the level of adenosine-5-triphosphate (ATP) synthesis, as compared with two different toxicity tests\\u000a based on inhibition of Daphnia magna mobility (NF EN ISO 6341) and inhibition of

Karine Estève; C. Poupot; P. Dabert; M. Mietton-Peuchot; V. Milisic



A global topology map of the Saccharomyces cerevisiae membrane proteome  

NASA Astrophysics Data System (ADS)

The yeast Saccharomyces cerevisiae is, arguably, the best understood eukaryotic model organism, yet comparatively little is known about its membrane proteome. Here, we report the cloning and expression of 617 S. cerevisiae membrane proteins as fusions to a C-terminal topology reporter and present experimentally constrained topology models for 546 proteins. By homology, the experimental topology information can be extended to 15,000 membrane proteins from 38 fully sequenced eukaryotic genomes. membrane proteins | membrane proteomics | yeast

Kim, Hyun; Melén, Karin; Österberg, Marie; von Heijne, Gunnar



Retrotransposon expression in ethanol-stressed Saccharomyces cerevisiae  

Microsoft Academic Search

There are five retrotransposon families in Saccharomyces cerevisiae, three (Ty1, Ty2, and Ty3) of which are known to be transcriptionally active. Early investigations reported yeast retrotransposons\\u000a to be stress-induced; however, microarray-based studies do not report retrotransposition-related Gene Ontology (GO) categories\\u000a in the ethanol stress response of S. cerevisiae. In this study, microarray technology was used to investigate the ethanol stress

Dragana Stanley; Sarah Fraser; Paul J. Chambers



Physiological effects of 5-hydroxymethylfurfural on Saccharomyces cerevisiae  

Microsoft Academic Search

The physiological effects of 5-hydroxymethylfurfural (HMF) on Saccharomyces cerevisiae CBS 8066 in the presence and absence of furfural were studied. Experiments were carried out by pulse addition of HMF (2–4?g\\/l)\\u000a as well as HMF (2?g\\/l) together with furfural (2?g\\/l) to batch cultivations of S. cerevisiae. Synthetic medium with glucose (50?g\\/l) as carbon and energy source was used. Addition of 4?g\\/l

M. J. Taherzadeh; L. Gustafsson; C. Niklasson; G. Lidén



Recovery of Saccharomyces cerevisiae from ethanol - induced growth inhibition  

SciTech Connect

Ethanol caused altered mobility of the lipophilic probe 1,6-diphenyl-1,3,5-hexatriene in plasma membrane preparations of Saccharomyces cerevisiae. Because lipids had been shown to protect yeast cells against ethanol toxicity, sterols, fatty acids, proteins, and combinations of these were tested; however, protection from growth inhibition was not seen. Ethanol-induced, prolonged lag periods and diminished growth rates in S. cerevisiae were reduced by an autoconditioning of the medium by the inoculum.

Walker-Caprioglio, H.M.; Rodriguez, R.J.; Parks, L.W.



Recovery of Saccharomyces cerevisiae from ethanol - induced growth inhibition  

Microsoft Academic Search

Ethanol caused altered mobility of the lipophilic probe 1,6-diphenyl-1,3,5-hexatriene in plasma membrane preparations of Saccharomyces cerevisiae. Because lipids had been shown to protect yeast cells against ethanol toxicity, sterols, fatty acids, proteins, and combinations of these were tested; however, protection from growth inhibition was not seen. Ethanol-induced, prolonged lag periods and diminished growth rates in S. cerevisiae were reduced by

H. M. Walker-Caprioglio; R. J. Rodriguez; L. W. Parks



Recovery of Saccharomyces cerevisiae from ethanol-induced growth inhibition.  

PubMed Central

Ethanol caused altered mobility of the lipophilic probe 1,6-diphenyl-1,3,5-hexatriene in plasma membrane preparations of Saccharomyces cerevisiae. Because lipids had been shown to protect yeast cells against ethanol toxicity, sterols, fatty acids, proteins, and combinations of these were tested; however, protection from growth inhibition was not seen. Ethanol-induced, prolonged lag periods and diminished growth rates in S. cerevisiae were reduced by an autoconditioning of the medium by the inoculum.

Walker-Caprioglio, H M; Rodriguez, R J; Parks, L W



Transcriptional changes associated with ethanol tolerance in Saccharomyces cerevisiae  

Microsoft Academic Search

Saccharomyces spp. are widely used for ethanol production; however, fermentation productivity is negatively affected by the impact of ethanol\\u000a accumulation on yeast metabolic rate and viability. This study used microarray and statistical two-way ANOVA analysis to compare\\u000a and evaluate gene expression profiles of two previously generated ethanol-tolerant mutants, CM1 and SM1, with their parent,\\u000a Saccharomyces cerevisiae W303-1A, in the presence

Dragana Stanley; Paul J. Chambers; Anthony Borneman; Sarah Fraser



Complete nucleotide sequence of Saccharomyces cerevisiae chromosome X.  

PubMed Central

The complete nucleotide sequence of Saccharomyces cerevisiae chromosome X (745 442 bp) reveals a total of 379 open reading frames (ORFs), the coding region covering approximately 75% of the entire sequence. One hundred and eighteen ORFs (31%) correspond to genes previously identified in S. cerevisiae. All other ORFs represent novel putative yeast genes, whose function will have to be determined experimentally. However, 57 of the latter subset (another 15% of the total) encode proteins that show significant analogy to proteins of known function from yeast or other organisms. The remaining ORFs, exhibiting no significant similarity to any known sequence, amount to 54% of the total. General features of chromosome X are also reported, with emphasis on the nucleotide frequency distribution in the environment of the ATG and stop codons, the possible coding capacity of at least some of the small ORFs (<100 codons) and the significance of 46 non-canonical or unpaired nucleotides in the stems of some of the 24 tRNA genes recognized on this chromosome. Images

Galibert, F; Alexandraki, D; Baur, A; Boles, E; Chalwatzis, N; Chuat, J C; Coster, F; Cziepluch, C; De Haan, M; Domdey, H; Durand, P; Entian, K D; Gatius, M; Goffeau, A; Grivell, L A; Hennemann, A; Herbert, C J; Heumann, K; Hilger, F; Hollenberg, C P; Huang, M E; Jacq, C; Jauniaux, J C; Katsoulou, C; Karpfinger-Hartl, L



Interaction of 6-phosphofructokinase with cytosolic proteins of Saccharomyces cerevisiae.  


Hetero-octameric 6-phosphofructokinase (Pfk-1) from Saccharomyces cerevisiae is composed of two types of subunits, alpha and beta, which are encoded by the unlinked genes PFK1 and PFK2. Pfk single deletion mutants expressing only one type of subunit exhibit Pfk-1 activity in vivo which, however, is completely lost immediately after cell disruption. In order to elucidate the preconditions of the in vivo activity of the mutant enzymes composed of either alpha- or beta-subunits, we have investigated their potential interaction with selected heat shock and cytoskeletal proteins, employing co-immunoprecipitation and immunofluorescence microscopy. Western blot analysis identified the mitochondrial chaperonin Hsp60, as well as the cytoskeleton proteins alpha-tubulin and actin, in complexes with Pfk-1 that were co-precipitated from a cell-free extract of a pfk2 single deletion mutant expressing only the alpha-subunit. The interaction of the corresponding mutant enzyme and Hsp60 was found to depend on the ATP concentration of the extract. Immunofluorescence microscopy displayed a conspicuously filamentous arrangement of the Pfk-1 mutant protein, exclusively in the pfk2 single deletion mutant. The analysis of structure and activity of Pfk-1 expressed in S. cerevisiae mutant strains defective in various heat shock proteins (TRiC/CCT, Hsp70, Hsp 104) and in the respective wild-type background did not reveal significant differences. PMID:15116431

Schwock, Jörg; Kirchberger, Jürgen; Edelmann, Anke; Kriegel, Thomas M; Kopperschläger, Gerhard



Studies of anaerobic and aerobic glycolysis in Saccharomyces cerevisiae  

Microsoft Academic Search

Glucose metabolism was followed in suspensions of Saccharomyces cerevisiae by using 13C NMR and 14C radioactive labeling techniques and by Warburg manometer experiments. These experiments were performed for cells grown with various carbon sources in the growth medium, so as to evaluate the effect of catabolite repression. The rate of glucose utilization was most conveniently determined by the 13C NMR

J. A. den Hollander; K. Ugurbil; T. R. Brown; M. Bednar; C. Redfield; R. G. Shulman



Identification of Saccharomyces cerevisiae Spindle Pole Body Remodeling Factors  

Microsoft Academic Search

The Saccharomyces cerevisiae centrosome or spindle pole body (SPB) is a dynamic structure that is remodeled in a cell cycle dependent manner. The SPB increases in size late in the cell cycle and during most cell cycle arrests and exchanges components during G1\\/S. We identified proteins involved in the remodeling process using a strain in which SPB remodeling is conditionally

Kristen B. Greenland; Huiming Ding; Michael Costanzo; Charles Boone; Trisha N. Davis



Size and Scar Distributions of the Yeast Saccharomyces Cerevisiae.  

National Technical Information Service (NTIS)

A model for the growth of populations of Saccharomyces cerevisiae is formulated and analyzed. The probability of bud emergence is assumed to depend on the size of the cell. Under certain conditions on birth size the model can be reduced to a single renewa...

M. Gyllenberg



Molecular Mechanism of Terbinafine Resistance in Saccharomyces cerevisiae  

Microsoft Academic Search

Ten mutants of the yeast Saccharomyces cerevisiae resistant to the antimycotic terbinafine were isolated after chemical or UV mutagenesis. Molecular analysis of these mutants revealed single base pair exchanges in the ERG1 gene coding for squalene epoxidase, the target of terbinafine. The mutants did not show cross-resistance to any of the substrates of various pleiotropic drug resistance efflux pumps tested.

Regina Leber; Sandra Fuchsbichler; V. Klobucnikova; Natascha Schweighofer; Eva Pitters; Kathrin Wohlfarter; Mojca Lederer; Karina Landl; Christoph Ruckenstuhl; Ivan Hapala; Friederike Turnowsky



Identification and Characterization of Phenylpyruvate Decarboxylase Genes in Saccharomyces cerevisiae  

Microsoft Academic Search

Received 9 December 2002\\/Accepted 7 May 2003 Catabolism of amino acids via the Ehrlich pathway involves transamination to the corresponding -keto acids, followed by decarboxylation to an aldehyde and then reduction to an alcohol. Alternatively, the aldehyde may be oxidized to an acid. This pathway is functional in Saccharomyces cerevisiae, since during growth in glucose-limited chemostat cultures with phenylalanine as

Zeynep Vuralhan; Marcos A. Morais; Siew-Leng Tai; Matthew D. W. Piper; Jack T. Pronk



Biolistic nuclear transformation of Saccharomyces cerevisiae and other fungi  

Microsoft Academic Search

Tungsten microprojectiles coated with nucleic acid and accelerated to velocities of approximately 500 m\\/s, can penetrate living cells and tissues with consequent expression of the introduced genes (Klein et al. 1987). Saccharomyces cerevisiae is used here as a model system to define the basic parameters governing the biolistic (biological-ballistic) delivery of DNA into cells. Among the physical factors affecting the

Daniele Armaleo; Guang-Ning Ye; Theodore M. Klein; Katherine B. Shark; John C. Sanford; Stephen Albert Johnston



Role of mitochondria in ethanol tolerance of Saccharomyces cerevisiae  

Microsoft Academic Search

The presence of active mitochondria and oxidative metabolism is shown to be essential to maintain low inhibition levels by ethanol of the growth rate (µ), fermentation rate (v) or respiration rate (?) of Saccharomyces cerevisiae wild type strain S288C. Cells which have respiratory metabolism show Ki (ethanol inhibition constant) values for µ, v and ?, higher (Ki>1 M) than those

Andrés Aguilera; Tahía Benítez



Production of lipid compounds in the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

This review describes progress using the yeast Saccharomyces cerevisiae as a model organism for the fast and efficient analysis of genes and enzyme activities involved in the lipid biosynthetic pathways of several donor organisms. Furthermore, we assess the impact of baker's yeast on the production of novel, high-value lipid compounds. Yeast can be genetically modified to produce selected substances in

M. Veen; C. Lang



Genome-Scale Reconstruction of the Saccharomyces cerevisiae Metabolic Network  

Microsoft Academic Search

The metabolic network in the yeast Saccharomyces cerevisiae was reconstructed using currently available genomic, biochemical, and physiological information. The metabolic reactions were compartmentalized between the cytosol and the mitochondria, and transport steps between the compartments and the environment were included. A total of 708 structural open reading frames (ORFs) were accounted for in the reconstructed network, corresponding to 1035 metabolic

Jochen Forster; Iman Famili; Patrick Fu; Bernhard Ø; Jens Nielsen



Complete Nucleotide Sequence of Saccharomyces cerevisiae Chromosome VIII  

Microsoft Academic Search

The complete nucleotide sequence of Saccharomyces cerevisiae chromosome VIII reveals that it contains 269 predicted or known genes (300 base pairs or larger). Fifty-nine of these genes (22 percent) were previously identified. Of the 210 novel genes, 65 are predicted to encode proteins that are similar to other proteins of known or predicted function. Sixteen genes appear to be relatively

M. Johnston; S. Andrews; R. Brinkman; J. Cooper; H. Ding; J. Dover; Z. Du; A. Favello; L. Fulton; S. Gattung; C. Geisel; J. Kirsten; T. Kucaba; L. Hillier; M. Jier; L. Johnston; Y. Langston; P. Latreille; E. J. Louis; C. Macri; E. Mardis; S. Menezes; L. Mouser; M. Nhan; L. Rifkin; L. Riles; H. St. Peter; E. Trevaskis; K. Vaughan; D. Vignati; L. Wilcox; P. Wohldman; R. Waterston; R. Wilson; M. Vaudin




Microsoft Academic Search

In commercial ethanol production producers often use sugar cane molasses as raw material due to their abundance and low costs. The most employed microorganisms used for fermentation is Saccharomyces cerevisiae yeasts due to their ability to hydrolyze sucrose from cane molasses into glucose and fructose, two easily assimilable hexoses. The aim of this study was to evaluate the application of



Continuous ethanol fermentation using immobilized yeast cells. [Saccharomyces cerevisiae  

Microsoft Academic Search

Growing cells of Saccharomyces cerevisiae immobilized in calcium alginate gel beads were employed in fluidized-bed reactors for continuous ethanol fermentation from cane molasses and other sugar sources. Some improvements were made in order to avoid microbial contamination and keep cell viability for stable long run operations. Notably, entrapment of sterol and unsaturated fatty acid into immobilized gel beads enhanced ethanol

M. Nagashima; M. Azuma; S. Noguchi; K. Inuzuka; H. Samejima



Transcriptional Response of Saccharomyces cerevisiae to Desiccation and Rehydration  

Microsoft Academic Search

A transcriptional analysis of the response of Saccharomyces cerevisiae strain BY4743 to controlled air-drying (desiccation) and subsequent rehydration under minimal glucose conditions was performed. Expression of genes involved in fatty acid oxidation and the glyoxylate cycle was observed to increase during drying and remained in this state during the rehydration phase. When the BY4743 expression profile for the dried sample

Jatinder Singh; Deept Kumar; Naren Ramakrishnan; Vibha Singhal; Jody Jervis; James F. Garst; Stephen M. Slaughter; Andrea M. DeSantis; Malcolm Potts; Richard F. Helm



Expression of Saccharomyces cerevisiae endochitinase in Aspergillus awamori  

Microsoft Academic Search

A gene encoding endochitinase activity was isolated by PCR from Saccharomyces cerevisiae NCYC686 and placed under the control of fungal transcriptional elements regulating glucoamylase expression. Following transformation of Aspergillus awamori with this expression construct, heterologous endochitinase was induced in positive transformants by the addition of starch to the growth medium. A series of optimisation and process development studies were then

Richard A. Murphy; Ronan F. G. Power



Analysis of the RNA Content of the Yeast "Saccharomyces Cerevisiae"  

ERIC Educational Resources Information Center

|In this article, the authors describe an interconnected set of relatively simple laboratory experiments in which students determine the RNA content of yeast cells and use agarose gel electrophoresis to separate and analyze the major species of cellular RNA. This set of experiments focuses on RNAs from the yeast "Saccharomyces cerevisiae", a…

Deutch, Charles E.; Marshall, Pamela A.



Inhibition of glycolysis by furfural in Saccharomyces cerevisiae  

Microsoft Academic Search

Furfural, a Maillard reaction product, was found to inhibit growth and alcohol production by Saccharomyces cerevisiae. Furfural concentrations above 1 mg ml-1 significantly decreased CO2 evolution by resuspended yeast cells. Important glycolytic enzymes such as hexokinase, phosphofructokinase, triosephosphate dehydrogenase, aldolase and alcohol dehydrogenase were assayed in presence of furfural. Dehydrogenases appeared to be the most sensitive enzymes and are probably

Nirupama Banerjee; Rakesh Bhatnagar; L. Viswanathan



Improving biomass sugar utilization by engineered Saccharomyces cerevisiae  

Technology Transfer Automated Retrieval System (TEKTRAN)

The efficient utilization of all available sugars in lignocellulosic biomass, which is more abundant than available commodity crops and starch, represents one of the most difficult technological challenges for the production of bioethanol. The well-studied yeast Saccharomyces cerevisiae has played a...


Stoichiometry and compartmentation of NADH metabolism in Saccharomyces cerevisiae  

Microsoft Academic Search

In Saccharomyces cerevisiae, reduction of NAD+ to NADH occurs in dissimilatory as well as in assimilatory reactions. This review discusses mechanisms for reoxidation of NADH in this yeast, with special emphasis on the metabolic compartmentation that occurs as a consequence of the impermeability of the mitochondrial inner membrane for NADH and NAD+. At least five mechanisms of NADH reoxidation exist

Barbara M. Bakker; Karin M. Overkamp; Antonius J. A. van Maris; Peter Kötter; Marijke A. H. Luttik; Johannes P. van Dijken; Jack T. Pronk



Atomic Force Microscopy Study of the Adhesion of Saccharomyces cerevisiae  

Microsoft Academic Search

An atomic force microscope (AFM) has been used to quantify directly the adhesion of metabolically active Saccharomyces cerevisiae cells at a hydrophilic mica surface, a mica surface with a hydrophobic coating, and a protein-coated mica surface in an aqueous environment. The measurements used “cell probes” constructed by immobilizing a single cell at the apex of a tipless AFM cantilever. Adhesion

W. Richard Bowen; Robert W. Lovitt; Chris J. Wright



Gene relics in the genome of the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

There is increasing evidence that DNA duplication is a common and ongoing process that plays a major role in molecular evolution of genomes and that a large fraction of the duplicated gene copies becomes non-functional by accumulation of deleterious mutations. In order to describe this phenomenon, we systematically searched the 6404 intergenic regions (IRs) of the genome of Saccharomyces cerevisiae

Ingrid Lafontaine; Gilles Fischer; Emmanuel Talla; Bernard Dujon



Polyamine metabolism in Saccharomyces cerevisiae exposed to ethanol  

Microsoft Academic Search

Growth of the yeast Saccharomyces cerevisiae was unaffected by up to 24 h exposure to ethanol concentrations ranging from 1% to 9%, but was reduced following exposure to 12% ethanol. Concentrations of the polyamines putrescine, cadaverine and spermidine were not affected by a 24 h exposure to 12% ethanol, although there was a significant increase in spermine level. These changes

Dale Walters; Tracy Cowley



Exclusion of Saccharomyces kudriavzevii from a wine model system mediated by Saccharomyces cerevisiae.  


This study investigated the competition and potential hybrid generation between the species Saccharomyces cerevisiae and S. kudriavzevii in a wine-model environment. Our main goal was to understand why S. kudriavzevii has not been found in wine fermentations whilst their hybrids are present. Auxotrophic mutants (Ura(-) and Lys(-)) were used to favour the selection of hybrids and to specifically differentiate the two species in mixed fermentations carried out at different temperatures (17 °C, 24 °C and 31 °C). Both yeasts showed a reduction in their maximum specific growth rates in mixed fermentations, indicating a clear antagonistic effect between the two microorganisms. Temperature played an important role in this competition. In this way, S. kudriavzevii was less affected at 17 °C, but S. cerevisiae was clearly the best competitor at 31 °C, preventing the growth of S. kudriavzevii. Population levels of S. kudriavzevii always significantly decreased in the presence of S. cerevisiae. Ethanol was measured throughout the fermentations and in all cases S. kudriavzevii growth was arrested when ethanol levels were < 5 g/l, indicating that this compound did not influence the competitive exclusion of S. kudriavzevii. Killer factors were also discarded due to the K(-) R(-) phenotype of both strains. Finally, no prototrophic interspecific hybrids were isolated in small-scale fermentations at any temperature assayed. Our results show that the lack of competitiveness exhibited by S. kudriavzevii, especially at high temperatures, explains the absence of this species in wine fermentations, suggesting that natural S. cerevisiae × S. kudriavzevii hybrids most likely originated in wild environments rather than in industrial fermentations. PMID:21381110

Arroyo-López, F Noé; Pérez-Través, Laura; Querol, Amparo; Barrio, Eladio



Genetic diversity study of the yeast Saccharomyces bayanus var. uvarum reveals introgressed subtelomeric Saccharomyces cerevisiae genes.  


Intraspecies polymorphism of the yeast Saccharomyces bayanus var. uvarum was studied using the polymerase chain reaction with a microsatellite primer (GTG)(5). Sixty-nine strains of different origins were analyzed. There existed a correlation between PCR patterns of the strains and the source of their isolation: the type of wine and the particular winemaking region. Southern hybridization analysis revealed for the first time introgression between Saccharomyces cerevisiae and S. bayanus var. uvarum. Two strains isolated from alcoholic beverages in Hungary and identified by genetic analysis as S. bayanus var. uvarum were found to harbor a number of S. cerevisiae subtelomeric sequences: Y', SUC, RTM and MAL. PMID:21112388

Naumova, Elena S; Naumov, Gennadi I; Michailova, Yulia V; Martynenko, Nikolay N; Masneuf-Pomarède, Isabelle



Bcl2 family members inhibit oxidative stress-induced programmed cell death in Saccharomyces cerevisiae  

Microsoft Academic Search

Selected antiapoptotic genes were expressed in baker’s yeast (Saccharomyces cerevisiae) to evaluate cytoprotective effects during oxidative stress. When exposed to treatments resulting in the generation of reactive oxygen species (ROS), including H2O2, menadione, or heat shock, wild-type yeast died and exhibited apoptotic-like characteristics, consistent with previous studies. Yeast strains were generated expressing nematode ced-9, human bcl-2, or chicken bcl-xl genes.

Shao-Rong Chen; David D Dunigan; Martin B Dickman



Serine racemase homologue of Saccharomyces cerevisiae has L- threo-3-hydroxyaspartate dehydratase activity  

Microsoft Academic Search

The NH2-terminal amino acid sequence of L-threo-3-hydroxyaspartate dehydratase from Pseudomonas sp. T62 showed significant similarity to that of the SRY1 gene product of Saccharomyces cerevisiae (serine racemase in yeast). SRY1 was cloned and expressed in Escherichia coli, and the gene product was purified and partially characterized. The SRY1 gene product exhibited dehydratase activity specific for L-threo-3-hydroxyaspartate (Km=3.9 mM, Vmax=110 ?mol

Masaru Wada; Shigeru Nakamori; Hiroshi Takagi



Evolutionary engineering of Saccharomyces cerevisiae for efficient aerobic xylose consumption.  


Industrial biotechnology aims to develop robust microbial cell factories, such as Saccharomyces cerevisiae, to produce an array of added value chemicals presently dominated by petrochemical processes. Xylose is the second most abundant monosaccharide after glucose and the most prevalent pentose sugar found in lignocelluloses. Significant research efforts have focused on the metabolic engineering of S. cerevisiae for fast and efficient xylose utilization. This study aims to metabolically engineer S. cerevisiae, such that it can consume xylose as the exclusive substrate while maximizing carbon flux to biomass production. Such a platform may then be enhanced with complementary metabolic engineering strategies that couple biomass production with high value-added chemical. Saccharomyces cerevisiae, expressing xylose reductase, xylitol dehydrogenase and xylulose kinase, from the native xylose-metabolizing yeast Pichia stipitis, was constructed, followed by a directed evolution strategy to improve xylose utilization rates. The resulting S. cerevisiae strain was capable of rapid growth and fast xylose consumption producing only biomass and negligible amount of byproducts. Transcriptional profiling of this strain was employed to further elucidate the observed physiology confirms a strongly up-regulated glyoxylate pathway enabling respiratory metabolism. The resulting strain is a desirable platform for the industrial production of biomass-related products using xylose as a sole carbon source. PMID:22487265

Scalcinati, Gionata; Otero, José Manuel; Van Vleet, Jennifer R H; Jeffries, Thomas W; Olsson, Lisbeth; Nielsen, Jens



Modelling neurodegeneration in Saccharomyces cerevisiae: why cook with baker's yeast?  


In ageing populations, neurodegenerative diseases increase in prevalence, exacting an enormous toll on individuals and their communities. Multiple complementary experimental approaches are needed to elucidate the mechanisms underlying these complex diseases and to develop novel therapeutics. Here, we describe why the budding yeast Saccharomyces cerevisiae has a unique role in the neurodegeneration armamentarium. As the best-understood and most readily analysed eukaryotic organism, S. cerevisiae is delivering mechanistic insights into cell-autonomous mechanisms of neurodegeneration at an interactome-wide scale. PMID:20424620

Khurana, Vikram; Lindquist, Susan



Lysis protein S of phage lambda functions in Saccharomyces cerevisiae.  

PubMed Central

The lambda S lysis gene was cloned into a Saccharomyces cerevisiae expression vector under GAL1 control. Induction with galactose in S. cerevisiae terminated cell growth and prevented colony formation. Several membrane proteins immunoreactive with anti-S antibody accumulated in the membranes, indicating that sodium dodecyl sulfate-resistant oligomers of S are formed, similar to those observed in the membranes of Escherichia coli cells killed by expression of the S gene. These observations suggest that the S gene product functions as a cytotoxic protein in the yeast cytoplasmic membrane as it does in the bacterial membrane. Images

Garrett, J; Bruno, C; Young, R



[Preliminary proteome analysis for Saccharomyces cerevisiae under different culturing conditions].  


For the investigation of the metabolic regulation of Saccharomyces cerevisiae under different culture conditions, the proteins of cell utilizing various carbon sources were separated by two-dimensional electrophoresis with immobilized pH gradients as the first dimension and SDS-PAGE as the second. Samples were taken in the log phase of batch culture using glucose or lactic acid as carbon source, while another sample was taken from the broth when glucose was consumed up and ethanol accumulated in the previous phase was further metabolized. After electrophoresis, the protein spots were detected by silver-stain in a Hoefer Automated Gel Stainer with a protein silver staining kit. Silver-stained gels were scanned and digitized to create computer images. About 500 protein spots were detected by employing the 2D proteome image analysis system Image Master 2D Elite and SWISS-2DPAGE proteome database. Most of the protein expressed and involved in the glycolysis, pentose phosphate (PP) pathway, anaplerotic pathway, as well as TCA cycle were analyzed. The metabolism regulation of protein level for Saccharomyces cerevisiae under various carbon sources, as well as during different phase of growth, was studied. The expression of several glycolytic enzymes (glk, pgi, pgk, eno, pyk) was up-regulated while the expression of enzymes in oxidative pentose phosphate pathway (zwf, gnd) was down-regulated when ethanol and lactic acid were taken as carbon source. Simultaneously, frucotose 1,6-biphosphatase was found to be up-regulated due to the gluconeogenic requirement. Citrate synthase and Malate dehydrogenase do not exhibit significant difference, indicating TCA cycle is necessary when utilizing glucose, ethanol or lactic acid as carbon source. Thus, the NADPH loss due to the repressed pentose phosphate pathway could be compensated by TCA cycle in cases of ethanol and lactic acid. The expression of malic enzyme and isocitrate lyase are activated to a large extent when metabolizing ethanol, indicating glyoxylate shunt is essential in transferring ethanol to generate four carbon precursors for the biosynthesis and the NADP-dependent malic enzyme could also serve as compensation mechanism for NADPH loss in this case. PMID:15971613

Zhang, Hui-Min; Yao, Shan-Jing; Peng, Li-Feng; Shimizu, Kazuyuki



Effects of moderate pressure on premeability and viability of Saccharomyces cerevisiae cells  

Microsoft Academic Search

With CO2 and N2 as the pressure media, the effects of the moderate pressure (0.1–1.0MPa) and the holding time on the conductivities of the\\u000a cell suspension of Saccharomyces cerevisiae CICC1447 and Saccharomyces cerevisiae CICC1339, as well as the absorbances of the supernatant (after centrifuged) at 280 nm (A280) and 260 nm (A260) were determined. The membrane permeability of Saccharomyces cerevisiae

Shi-Ru Jia; Na-Chen; Yu-Jie Dai; Chang-Sheng Qiao; Jian-Dong Cui; Bo-Ning Liu



Genotoxicity assessment of amaranth and allura red using Saccharomyces cerevisiae.  


Amaranth (E123) and Allura red (E129), very important food azo dyes used in food, drug, paper, cosmetic and textile industries, were assessed for their genotoxic potential through comet assay in yeast cells. Comet assay was standardized by with different concentration of H(2)O(2). Concentrations of Amaranth and Allura red were maintained in sorbitol buffer starting from 9.76 to 5,000 ?g/mL and 1 × 10(4) cells were incubated at two different incubation temperatures 28 and 37°C. Amaranth (E123) and Allura red (E129) were found to exhibit their genotoxic effect directly in Saccharomyces cerevisiae. No significant genotoxic activity was observed for Amaranth and Allura red at 28°C but at 37°C direct relation of Amaranth concentration with comet tail was significant and no positive relation was seen with time exposure factor. At 37°C the minimum concentration of Amaranth and Allura red at which significant DNA damage observed through comet assay was 1,250 ?g/mL in 2nd h post exposure time. The results indicated that food colors should be carefully used in baking products as heavy concentration of food colors could affect the fermentation process of baking. PMID:23132362

Jabeen, Hafiza Sumara; ur Rahman, Sajjad; Mahmood, Shahid; Anwer, Sadaf



Distribution and regulation of stochasticity and plasticity in Saccharomyces cerevisiae  

NASA Astrophysics Data System (ADS)

Stochasticity is an inherent feature of complex systems with nanoscale structure. In such systems information is represented by small collections of elements (e.g., a few electrons on a quantum dot), and small variations in the populations of these elements may lead to big uncertainties in the information. Unfortunately, little is known about how to work within this inherently noisy environment to design robust functionality into complex nanoscale systems. Here, we look to the biological cell as an intriguing model system where evolution has mediated the trade-offs between fluctuations and function, and in particular we look at the relationships and trade-offs between stochastic and deterministic responses in the gene expression of budding yeast (Saccharomyces cerevisiae). We find gene regulatory arrangements that control the stochastic and deterministic components of expression, and show that genes that have evolved to respond to stimuli (stress) in the most strongly deterministic way exhibit the most noise in the absence of the stimuli. We show that this relationship is consistent with a bursty two-state model of gene expression, and demonstrate that this regulatory motif generates the most uncertainty in gene expression when there is the greatest uncertainty in the optimal level of gene expression.

Dar, R. D.; Karig, D. K.; Cooke, J. F.; Cox, C. D.; Simpson, M. L.



Regulation of phospholipid synthesis in Saccharomyces cerevisiae by zinc.  


Zinc is an essential nutrient required for the growth and metabolism of eukaryotic cells. In this work, we examined the effects of zinc depletion on the regulation of phospholipid synthesis in the yeast Saccharomyces cerevisiae. Zinc depletion resulted in a decrease in the activity levels of the CDP-diacylglycerol pathway enzymes phosphatidylserine synthase, phosphatidylserine decarboxylase, phosphatidylethanolamine methyltransferase, and phospholipid methyltransferase. In contrast, the activity of phosphatidylinositol synthase was elevated in response to zinc depletion. The level of Aut7p, a marker for the induction of autophagy, was also elevated in zinc-depleted cells. For the CHO1-encoded phosphatidylserine synthase, the reduction in activity in response to zinc depletion was controlled at the level of transcription. This regulation was mediated through the UAS(INO) element and by the transcription factors Ino2p, Ino4p, and Opi1p that are responsible for the inositol-mediated regulation of UAS(INO)-containing genes involved in phospholipid synthesis. Analysis of the cellular composition of the major membrane phospholipids showed that zinc depletion resulted in a 66% decrease in phosphatidylethanolamine and a 29% increase in phosphatidylinositol. A zrt1Delta zrt2Delta mutant (defective in the plasma membrane zinc transporters Zrt1p and Zrt2p) grown in the presence of zinc exhibited a phospholipid composition similar to that of wild type cells depleted for zinc. These results indicated that a decrease in the cytoplasmic levels of zinc was responsible for the alterations in phospholipid composition. PMID:15028711

Iwanyshyn, Wendy M; Han, Gil-Soo; Carman, George M



Saccharomyces cerevisiae: a sexy yeast with a prion problem.  


Yeast prions are infectious proteins that spread exclusively by mating. The frequency of prions in the wild therefore largely reflects the rate of spread by mating counterbalanced by prion growth slowing effects in the host. We recently showed that the frequency of outcross mating is about 1% of mitotic doublings with 23-46% of total matings being outcrosses. These findings imply that even the mildest forms of the [PSI+], [URE3] and [PIN+] prions impart > 1% growth/survival detriment on their hosts. Our estimate of outcrossing suggests that Saccharomyces cerevisiae is far more sexual than previously thought and would therefore be more responsive to the adaptive effects of natural selection compared with a strictly asexual yeast. Further, given its large effective population size, a growth/survival detriment of > 1% for yeast prions should strongly select against prion-infected strains in wild populations of Saccharomyces cerevisiae. PMID:23764836

Kelly, Amy C; Wickner, Reed B


Ethanol production by Saccharomyces cerevisiae in biofilm reactors  

Microsoft Academic Search

  Biofilms are natural forms of cell immobilization in which microorganisms attach to solid supports. At ISU, we have developed\\u000a plastic composite-supports (PCS) (agricultural material (soybean hulls or oat hulls), complex nutrients, and polypropylene)\\u000a which stimulate biofilm formation and which supply nutrients to the attached microorganisms. Various PCS blends were initially\\u000a evaluated in repeated-batch culture-tube fermentation with Saccharomyces cerevisiae (ATCC 24859)

A Demirci; A L Pometto III; K-L G Ho



Posttranscriptional regulation in the myo1? mutant of Saccharomyces cerevisiae  

Microsoft Academic Search

BACKGROUND: Saccharomyces cerevisiae myosin type II-deficient (myo1?) strains remain viable and divide, despite the absence of a cytokinetic ring, by activation of the PKC1-dependent cell wall integrity pathway (CWIP). Since the myo1? transcriptional fingerprint is a subset of the CWIP fingerprint, the myo1? strain may provide a simplified paradigm for cell wall stress survival. RESULTS: To explore the post-transcriptional regulation

Marielis E Rivera-Ruiz; José F Rodríguez-Quiñones; Pearl Akamine; José R Rodríguez-Medina



Transcriptional Remodeling in Response to Iron Deprivation in Saccharomyces cerevisiae  

Microsoft Academic Search

The budding yeast Saccharomyces cerevisiae responds to depletion of iron in the environment by activating Aft1p, the major iron-dependent transcription factor, and by transcribing systems involved in the uptake of iron. Here, we have studied the transcriptional response to iron deprivation and have identified new Aft1p target genes. We find that other metabolic pathways are regulated by iron: biotin uptake

Minoo Shakoury-Elizeh; John Tiedeman; Jared Rashford; Tracey Ferea; Janos Demeter; Emily Garcia; Ronda Rolfes; Patrick O. Brown; David Botstein; Caroline C. Philpott



Response to different oxidants of Saccharomyces cerevisiae ure2? mutant  

Microsoft Academic Search

Growth of Saccharomyces cerevisiae\\u000a ure2? mutant strain was investigated in the presence of diverse oxidant compounds. The inability of the strain to grow on a medium\\u000a supplemented with H2O2 was confirmed and a relationship between diminishing levels of glutathione (GSH) and peroxide sensitivity was established.\\u000a Data for the lack of significant effect of URE2 disruption on the cellular growth in

Tatina T. Todorova; Ventsislava Y. Petrova; Stéphane Vuilleumier; Anna V. Kujumdzieva



DOT4 Links Silencing and Cell Growth in Saccharomyces cerevisiae  

Microsoft Academic Search

Transcriptional silencing in Saccharomyces cerevisiae occurs at specific loci and is mediated by a multiprotein complex that includes Rap1p and the Sir proteins. We studied the function of a recently identified gene, DOT4, that disrupts silencing when overexpressed. DOT4 encodes an ubiquitin processing protease (hydrolase) that is primarily located in the nucleus. By two-hybrid analysis, the amino-terminal third of Dot4p



Promotion of sporulation by caffeine pretreatment in Saccharomyces cerevisiae  

Microsoft Academic Search

Changes in RNase activity during sporulation of a homothallic diploid strain of Saccharomyces cerevisiae were measured in caffeine-treated and non-treated cells.1.In caffeine-treated cells soon after the transfer to the sporulation medium a significant increase in RNase activity was observed; in control cells the rise of RNase activity was less and started after a lag period of 5 h. The final

Michio Tsuboi; Naohiko Yanagishima



Effects of growth conditions on mitochondrial morphology in Saccharomyces cerevisiae  

Microsoft Academic Search

Effects of growth conditions on mitochondrial morphology were studied in livingSaccharomyces cerevisiae cells by vital staining with the fluorescent dye dimethyl-aminostyryl-methylpyridinium iodine (DASPMI), fluorescence microscopy, and confocal-scanning laser microscopy. Cells from respiratory, ethanol-grown batch cultures contained a large number of small mitochondria. Conversely, cells from glucose-grown batch cultures, in which metabolism was respiro-fermentative, contained small numbers of large, branched mitochondria.

Wiebe Visser; Edwin A. van Spronsen; Nanne Nanninga; Jack T. Pronk; J. Gijs Kuenen; Johannes R van Dijken



Characterizing the effect of nitrosative stress in Saccharomyces cerevisiae  

Microsoft Academic Search

Nitrosative stress has various pathophysiological implications. We here present a detailed characterization on the effect of nitrosative stress in Saccharomyces cerevisiae wild-type (Y190) and its isogenic flavohemoglobin mutant (?yhb1) strain grown in presence of non fermentable carbon source. On addition of sub-toxic dose of nitrosating agent both the strains showed microbiostatic effect. Cellular respiration was found to be significantly affected

Arindam Bhattacharjee; Uddalak Majumdar; Debasis Maity; Tuhin Subhra Sarkar; Achintya Mohan Goswami; Rupam Sahoo; Sanjay Ghosh



Global landscape of protein complexes in the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

Identification of protein–protein interactions often provides insight into protein function, and many cellular processes are performed by stable protein complexes. We used tandem affinity purification to process 4,562 different tagged proteins of the yeast Saccharomyces cerevisiae. Each preparation was analysed by both matrix-assisted laser desorption\\/ionization–time of flight mass spectrometry and liquid chromatography tandem mass spectrometry to increase coverage and accuracy.

Nevan J. Krogan; Gerard Cagney; Haiyuan Yu; Gouqing Zhong; Xinghua Guo; Alexandr Ignatchenko; Joyce Li; Shuye Pu; Nira Datta; Aaron P. Tikuisis; Thanuja Punna; José M. Peregrín-Alvarez; Michael Shales; Xin Zhang; Michael Davey; Mark D. Robinson; Alberto Paccanaro; James E. Bray; Anthony Sheung; Bryan Beattie; Dawn P. Richards; Veronica Canadien; Atanas Lalev; Frank Mena; Peter Wong; Andrei Starostine; Myra M. Canete; James Vlasblom; Samuel Wu; Chris Orsi; Sean R. Collins; Shamanta Chandran; Robin Haw; Jennifer J. Rilstone; Kiran Gandi; Natalie J. Thompson; Gabe Musso; Peter St Onge; Shaun Ghanny; Mandy H. Y. Lam; Gareth Butland; Amin M. Altaf-Ul; Shigehiko Kanaya; Ali Shilatifard; Erin O'Shea; Jonathan S. Weissman; C. James Ingles; Timothy R. Hughes; John Parkinson; Mark Gerstein; Shoshana J. Wodak; Andrew Emili; Jack F. Greenblatt



Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan  

Microsoft Academic Search

In diverse organisms, calorie restriction slows the pace of ageing and increases maximum lifespan. In the budding yeast Saccharomyces cerevisiae, calorie restriction extends lifespan by increasing the activity of Sir2 (ref. 1), a member of the conserved sirtuin family of NAD+-dependent protein deacetylases. Included in this family are SIR-2.1, a Caenorhabditis elegans enzyme that regulates lifespan, and SIRT1, a human

Konrad T. Howitz; Kevin J. Bitterman; Haim Y. Cohen; Dudley W. Lamming; Siva Lavu; Jason G. Wood; Robert E. Zipkin; Phuong Chung; Anne Kisielewski; Li-Li Zhang; Brandy Scherer; David A. Sinclair



Calorie restriction extends Saccharomyces cerevisiae lifespan by increasing respiration  

Microsoft Academic Search

Calorie restriction (CR) extends lifespan in a wide spectrum of organisms and is the only regimen known to lengthen the lifespan of mammals. We established a model of CR in budding yeast Saccharomyces cerevisiae. In this system, lifespan can be extended by limiting glucose or by reducing the activity of the glucose-sensing cyclic-AMP-dependent kinase (PKA). Lifespan extension in a mutant

Su-Ju Lin; Matt Kaeberlein; Alex A. Andalis; Lori A. Sturtz; Pierre-Antoine Defossez; Valeria C. Culotta; Gerald R. Fink; Leonard Guarente



Isolierung und Charakterisierung zweier Nucleasen aus Backhefe (Saccharomyces cerevisiae)  

Microsoft Academic Search

Summary Tworibo nucleases have been isolated from baker's yeast (Saccharomyces cerevisiae) and enriched 250 and 400 fold (fractionation by precipitation with acetone and ammonium sulfate followed by treating with tricalciumphosphate gel and ion exchange chromatography on DEAE-cellulose). Both enzymes are not active against bis-(p-nitrophenyl)-phosphat and cyclic phosphates and there is no phosphortransferase activity. Polynucleotides are splitted at the midth of

Diethard Dauber



Ergosterol production from molasses by genetically modified Saccharomyces cerevisiae  

Microsoft Academic Search

Ergosterol is an economically important metabolite produced by fungi. Recombinant Saccharomyces cerevisiae YEH56(pHXA42) with increased capacity of ergosterol formation was constructed by combined overexpression of sterol C-24(28)\\u000a reductase and sterol acyltransferase in the yeast strain YEH56. The production of ergosterol by this recombinant strain using\\u000a cane molasses (CM) as an inexpensive carbon source was investigated. An ergosterol content of 52.6 mg\\/g

Xiuping He; Xuena Guo; Nan Liu; Borun Zhang



Electrophysiology in the eukaryotic model cell Saccharomyces cerevisiae  

Microsoft Academic Search

Since the mid-1980s, use of the budding yeast, Saccharomyces cerevisiae, for expression of heterologous (foreign) genes and proteins has burgeoned for several major purposes, including facile genetic\\u000a manipulation, large-scale production of specific proteins, and preliminary functional analysis. Expression of heterologous\\u000a membrane proteins in yeast has not kept pace with expression of cytoplasmic proteins for two principal reasons: (1) although plant

Adam Bertl; Hermann Bihler; Carsten Kettner; Clifford L. Slayman



Hydrolysis and fermentation of amorphous cellulose by recombinant Saccharomyces cerevisiae  

Microsoft Academic Search

In this study, we expressed two cellulase encoding genes, an endoglucanase of Trichoderma reesei (EGI) and the ?-glucosidase of Saccharomycopsis fibuligera (BGL1), in combination in Saccharomyces cerevisiae. The resulting strain was able to grow on phosphoric acid swollen cellulose (PASC) through simultaneous production of sufficient extracellular endoglucanase and ?-glucosidase activity. Anaerobic growth (0.03h?1) up to 0.27gl?1 DCW was observed on

Riaan Den Haan; Shaunita H. Rose; Lee R. Lynd; Willem H. van Zyl



Interaction Between Mismatch Repair and Genetic Recombination in Saccharomyces cerevisiae  

Microsoft Academic Search

The yeast Saccharomyces cerevisiae encodes a set of genes that show strong amino acid sequence simi- larity to MutS and MutL, proteins required for mismatch repair in Escherichia coli. We examined the role of MSH2 and PMSl, yeast homologs of mutS and mutL, respectively, in the repair of base pair mismatches formed during meiotic recombination. By using specifically marked HIS4

Eric Alani; Robert A. G. Reenan; Richard D. Kolodner



Preparation of Cell-Free Splicing Extracts from Saccharomyces cerevisiae.  


Much of our understanding of the mechanism of splicing comes from the analysis of cell extracts able to carry out splicing complex formation and splicing reactions in vitro using exogenously added synthetic model pre-mRNA transcripts. This protocol describes the preparation of whole-cell extracts from the budding yeast Saccharomyces cerevisiae. These extracts can be used to dissect the biochemical steps of the splicing reaction and to determine the macromolecules, cofactors, and substrate features necessary for successful splicing. PMID:24086052

Ares, Manuel



SACCHAROMYCES CEREVISIAE Recessive Suppressor That Circumvents Phosphatidylserine Deficiency  

PubMed Central

Phenotypic reversion of six independent Saccharomyces cerevisiae cho1 mutants was shown to be due predominantly to mutation of an unlinked gene, eam1. The eam1 gene was located very close to ino1 on chromosome X by meiotic tetrad analysis. Recessive eam1 mutations did not correct the primary cho1 defect in phosphatidylserine synthesis but made endogenous ethanolamine available for sustained nitrogenous phospholipid synthesis. A novel biochemical contribution to nitrogenous lipid synthesis is indicated by the eam1 mutants.

Atkinson, Katharine D.



D-xylulose fermentation to ethanol by Saccharomyces cerevisiae  

Microsoft Academic Search

Commercial bakers' yeast (Saccharomyces cerevisiae) was used to study the conversion of D-xylulose to ethanol in the presence of D-xylose. The rate of ethanol production increased with an increase in yeast cell density. The optimal temperature for D-xylulose fermentation was 35 degrees Celcius, and the optimal pH range was 4 to 6. The fermentation of D-xylulose by yeast resulted in

L. C. Chiang; C. S. Gong; L. F. Chen; G. T. Tsao



A Mutant of Saccharomyces cerevisiae Defective for Nuclear Fusion  

Microsoft Academic Search

A mutant unable to fuse nuclei during mating has been isolated from standard wild-type Saccharomyces cerevisiae. Tetrad analysis of the mutation responsible for this defect (kar1-1) shows that it segregates as a single Mendelian factor. The defect in kar1-1 appears to be nuclear limited. Cytological and genetic evidence shows that in this mutant the events associated with zygote formation are

Jaime Conde; Gerald R. Fink



Characterization of Saccharomyces cerevisiae deficient in expression of phospholipase D.  

PubMed Central

A gene encoding phospholipase D (PLD) in Saccharomyces cerevisiae was identified. The 195 kDa product of PLD1 has 24% overall sequence identity with a plant PLD. Expression of yeast PLD activity was eliminated by one-step gene disruption. Yeast haploids lacking PLD activity were deficient in growth on non-fermentable carbon sources. Diploids lacking expression of PLD1 were unable to sporulate.

Ella, K M; Dolan, J W; Qi, C; Meier, K E



Exploring improved endoglucanase expression in Saccharomyces cerevisiae strains  

Microsoft Academic Search

The endoglucanase I and II genes (egI or Cel7B and egII or Cel5A) of Trichoderma reesei QM6a were successfully cloned and expressed in Saccharomyces cerevisiae under the transcriptional control of the yeast ENO1 promoter and terminator sequences. Random mutagenesis of the egI-bearing plasmid resulted in a twofold increase in extracellular EGI activity. Both endoglucanase genes were co-expressed\\u000a with the synthetic,

Lisa du Plessis; Shaunita H. Rose; Willem H. van Zyl



Postreplication Repair Inhibits CAG · CTG Repeat Expansions in Saccharomyces cerevisiae  

Microsoft Academic Search

Trinucleotide repeats (TNRs) are unique DNA microsatellites that can expand to cause human disease. Recently, Srs2 was identified as a protein that inhibits TNR expansions in Saccharomyces cerevisiae. Here, we demonstrate that Srs2 inhibits CAG · CTG expansions in conjunction with the error-free branch of postrep- lication repair (PRR). Like srs2 mutants, expansions are elevated in rad18 and rad5 mutants,

Danielle L. Daee; Tony Mertz; Robert S. Lahue


Distribution and correlation of three oenological traits in Saccharomyces cerevisiae  

Microsoft Academic Search

The improvement of yeast starters for wine making, with classical genetic techniques, relays on the effective independence\\u000a of the main oenological traits in order to combine them optimally. The analysis of three characters (ethanol production, volatile\\u000a acidity and fermenting rate) in 787Saccharomyces cerevisiae strains, isolated from several parts of the world and substrates, shows moderate correlation between the volatile acidity

Laura Corte; Paolo Rellini; Francesco Sciascia; Raffaele De Nicola; Fabrizio Fatichenti; Gianluigi Cardinali



A rapid preparation of plasmid DNA from Saccharomyces cerevisiae  

Microsoft Academic Search

A procedure for extraction of plasmid DNA fromSaccharomyces cerevisiae is described. The plasmid DNA of interest is extracted together with 2-?m circular DNA naturally occurring in many yeast\\u000a strains. Spheroplasts are lyzed at alkaline pH which denatures linear but not covalently closed circular (CCC) DNA. The CCC\\u000a DNA is recovered by ethanol precipitation and can be detected by gel electrophoresis

J. Brozmanová; Z. Holinová



Molecular and biochemical analysis of Saccharomyces cerevisiae cox1 mutants  

Microsoft Academic Search

We report on the molecular and biochemical analysis of a set of 13 respiratory deficient mutants of Saccharomyces cerevisiae which are specifically altered in COX1, the gene encoding the subunit Cox1p of cytochrome c oxidase. DNA sequence analysis shows that three are due to frameshift mutations, two to nonsense mutations, and eight to\\u000a missense mutations. All, except the missense mutant

Claire Lemaire; Sylviane Robineau; Pierre Netter



Redox Interactions between Saccharomyces cerevisiae and Saccharomyces uvarum in Mixed Culture under Enological Conditions  

PubMed Central

Wine yeast starters that contain a mixture of different industrial yeasts with various properties may soon be introduced to the market. The mechanisms underlying the interactions between the different strains in the starter during alcoholic fermentation have never been investigated. We identified and investigated some of these interactions in a mixed culture containing two yeast strains grown under enological conditions. The inoculum contained the same amount (each) of a strain of Saccharomyces cerevisiae and a natural hybrid strain of S. cerevisiae and Saccharomyces uvarum. We identified interactions that affected biomass, by-product formation, and fermentation kinetics, and compared the redox ratios of monocultures of each strain with that of the mixed culture. The redox status of the mixed culture differed from that of the two monocultures, showing that the interactions between the yeast strains involved the diffusion of metabolite(s) within the mixed culture. Since acetaldehyde is a potential effector of fermentation, we investigated the kinetics of acetaldehyde production by the different cultures. The S. cerevisiae-S. uvarum hybrid strain produced large amounts of acetaldehyde for which the S. cerevisiae strain acted as a receiving strain in the mixed culture. Since yeast response to acetaldehyde involves the same mechanisms that participate in the response to other forms of stress, the acetaldehyde exchange between the two strains could play an important role in inhibiting some yeast strains and allowing the growth of others. Such interactions could be of particular importance in understanding the ecology of the colonization of complex fermentation media by S. cerevisiae.

Cheraiti, Naoufel; Guezenec, Stephane; Salmon, Jean-Michel



A Saccharomyces cerevisiae-based bioassay for assessing pesticide toxicity.  


This study evaluates the toxic effect of three pesticides (Azoxystrobin, Cymoxanil, and Diuron) on the yeast Saccharomyces cerevisiae for the development of a new bioassay based on inhibition of S. cerevisiae metabolic activity at the level of adenosine-5-triphosphate (ATP) synthesis, as compared with two different toxicity tests based on inhibition of Daphnia magna mobility (NF EN ISO 6341) and inhibition of Vibrio fisheri activity (NF EN ISO 11348). The S. cerevisiae bioassay is cheaper and 96 times faster than the D. magna toxicity bioassay, but has lower sensitivity. It is as fast as the V. fisheri bioassay and more sensitive. Thus, this new toxicity test can be proposed for rapid detection of pesticide residues in environmental samples as a complement to the more expensive and time-consuming D. magna toxicity test. PMID:19856193

Estève, Karine; Poupot, C; Dabert, P; Mietton-Peuchot, M; Milisic, V



Genetic engineering of industrial strains of Saccharomyces cerevisiae.  


Genetic engineering has been successfully applied to Saccharomyces cerevisiae laboratory strains for different purposes: extension of substrate range, improvement of productivity and yield, elimination of by-products, improvement of process performance and cellular properties, and extension of product range. The potential of genetically engineered yeasts for the massive production of biofuels as bioethanol and other nonfuel products from renewable resources as lignocellulosic biomass hydrolysates has been recognized. For such applications, robust industrial strains of S. cerevisiae have to be used. Here, some relevant genetic and genomic characteristics of industrial strains are discussed in relation to the problematic of the genetic engineering of such strains. General molecular tools applicable to the manipulation of S. cerevisiae industrial strains are presented and examples of genetically engineered industrial strains developed for the production of bioethanol from lignocellulosic biomass are given. PMID:22160914

Le Borgne, Sylvie



Antimutagenic and antioxidant activity of Lisosan G in Saccharomyces cerevisiae.  


In the present study the antimutagenic and antioxidant effects of a powder of grain (Lisosan G) in yeast Saccharomyces cerevisiae were studied. Results showed that Lisosan G treatment decreased significantly the intracellular ROS concentration and mutagenesis induced by hydrogen peroxide in S. cerevisiae D7 strain. The effect of Lisosan G was then evaluated by using superoxide dismutase (SOD) proficient and deficient strains of S. cerevisiae. Lisosan G showed protective activity in sod1? and sod2? mutant strains, indicating an in vivo antioxidant effect. A high radical scavenging activity of Lisosan G was also demonstrated in vitro using the oxygen radical absorbance capacity (ORAC) assay. The obtained results showed a protective effect of Lisosan G in yeast cells, indicating that its antioxidant capacity contributes to its antimutagenic action. PMID:22953954

Frassinetti, Stefania; Della Croce, Clara Maria; Caltavuturo, Leonardo; Longo, Vincenzo



Overproduction of threonine by Saccharomyces cerevisiae mutants resistant to hydroxynorvaline.  

PubMed Central

In this work, we isolated and characterized mutants that overproduce threonine from Saccharomyces cerevisiae. The mutants were selected for resistance to the threonine analog alpha-amino-beta-hydroxynorvalerate (hydroxynorvaline), and, of these, the ones able to excrete threonine to the medium were chosen. The mutant strains produce between 15 and 30 times more threonine than the wild type does, and, to a lesser degree, they also accumulate isoleucine. Genetic and biochemical studies have revealed that the threonine overproduction is, in all cases studied, associated with the presence in the strain of a HOM3 allele coding for a mutant aspartate kinase that is totally or partially insensitive to feedback inhibition by threonine. This enzyme seems, therefore, to be crucial in the regulation of threonine biosynthesis in S. cerevisiae. The results obtained suggest that this strategy could be efficiently applied to the isolation of threonine-overproducing strains of yeasts other than S. cerevisiae, even those used industrially.

Ramos, C; Calderon, I L



Genome-Scale Reconstruction of the Saccharomyces cerevisiae Metabolic Network  

PubMed Central

The metabolic network in the yeast Saccharomyces cerevisiae was reconstructed using currently available genomic, biochemical, and physiological information. The metabolic reactions were compartmentalized between the cytosol and the mitochondria, and transport steps between the compartments and the environment were included. A total of 708 structural open reading frames (ORFs) were accounted for in the reconstructed network, corresponding to 1035 metabolic reactions. Further, 140 reactions were included on the basis of biochemical evidence resulting in a genome-scale reconstructed metabolic network containing 1175 metabolic reactions and 584 metabolites. The number of gene functions included in the reconstructed network corresponds to ?16% of all characterized ORFs in S. cerevisiae. Using the reconstructed network, the metabolic capabilities of S. cerevisiae were calculated and compared with Escherichia coli. The reconstructed metabolic network is the first comprehensive network for a eukaryotic organism, and it may be used as the basis for in silico analysis of phenotypic functions. [Supplemental material is available online at The detailed genome-scale reconstructed model of Saccharomyces cerevisiae can be found at or

Forster, Jochen; Famili, Iman; Fu, Patrick; Palsson, Bernhard ?.; Nielsen, Jens



Understanding the mechanism of heat stress tolerance caused by high trehalose accumulation in Saccharomyces cerevisiae using DNA microarray.  


DNA microarray analysis was performed to examine the stress tolerance mechanism of a Saccharomyces cerevisiae recombinant strain exhibiting high trehalose accumulation and heat stress tolerance. Results suggest that the upregulation of sugar transporter genes is one of the key events for heat stress tolerance of the recombinant strain. PMID:22222142

Mahmud, Siraje Arif; Hirasawa, Takashi; Furusawa, Chikara; Yoshikawa, Katsunori; Shimizu, Hiroshi



Enhancing beta-carotene production in Saccharomyces cerevisiae by metabolic engineering.  


Beta-carotene is known to exhibit a number of pharmacological and nutraceutical benefits to human health. Metabolic engineering of beta-carotene biosynthesis in Saccharomyces cerevisiae has been attracting the interest of many researchers. A previous work has shown that S. cerevisiae successfully integrated with phytoene synthase (crtYB) and phytoene desaturase (crtI) from Xanthophyllomyces dendrorhous could produce beta-carotene. In the present study, we achieved around 200% improvement in beta-carotene production in S. cerevisiae through specific site optimization of crtI and crtYB, in which five codons of crtI and eight codons of crtYB were rationally mutated. Furthermore, the effects of the truncated HMG-CoA reductase (tHMG1) from S. cerevisiae and HMG-CoA reductase (mva) from Staphylococcus aureus on the production of beta-carotene in S. cerevisiae were also evaluated. Our results indicated that mva from a prokaryotic organism might be more effective than tHMG1 for beta-carotene production in S. cerevisiae. PMID:23718229

Li, Qian; Sun, Zhiqiang; Li, Jing; Zhang, Yansheng



Ultrastructural changes of Saccharomyces cerevisiae in response to ethanol stress.  


In the fermentative process using Saccharomyces cerevisiae to produce bioethanol, the performance of cells is often compromised by the accumulation of ethanol. However, the mechanism of how S. cerevisiae responds against ethanol stress remains elusive. In the current study, S. cerevisiae cells were cultured in YPD (yeast extract - peptone - dextrose) medium containing various concentrations of ethanol (0%, 2.5%, 5%, 7.5%, 10%, and 15% (v/v)). Compared with the control group without ethanol, the mean cell volume of S. cerevisiae decreased significantly in the presence of 7.5% and 10% ethanol after incubation for 16 h (P < 0.05), and in the presence of 15% ethanol at all 3 sampling time points (1, 8, and 16 h) (P < 0.05). The exposure of S. cerevisiae cells to ethanol also led to an increase in malonyldialdehyde content (P < 0.05) and a decrease in sulfhydryl group content (P < 0.05). Moreover, the observations through transmission electron microscopy enabled us to relate ultrastructural changes elicited by ethanol with the cellular stress physiology. Under ethanol stress, the integrity of the cell membrane was compromised. The swelling or distortion of mitochondria together with the occurrence of a single and large vacuole was correlated with the addition of ethanol. These results suggested that the cell membrane is one of the targets of ethanol, and the degeneration of mitochondria promoted the accumulation of intracellular reactive oxygen species. PMID:24011341

Ma, Manli; Han, Pei; Zhang, Ruimin; Li, Hao



Ethanol production by immobilized Saccharomyces cerevisiae, Saccharomyces uvarum, and Zymomonas mobilis  

SciTech Connect

Saccharomyces cerevisiae NRRL Y-2034, Saccharomyces uvarum NRRL Y-1347, and Zymomonas mobilis NRRL B-806 each were separately immobilized in a Ca-alginate matrix and incubated in the presence of a free-flowing and continuous 1,3,5, 10, or 20% (w/w) glucose solution. In general, the yeast cells converted 100% of the 1,3, and 5% glucose to alcohol within 48 h and maintained such a conversion rate for at least two weeks. The bacterium converted ca. 90% (w/w) of the 1,3, and 5% glucose to alcohol continuously for one week. However, both the yeast and bacterium were inhibited in the highest glucose (20% w/w) solution. All of the immobilized cultures produced some alcohol for at least 14 days. Immobilized Saccharomyces cerevisiae was the best alcohol producer of all of the glucose concentrations; the yeast yielded 4.7 g ethanol/100 g solution within 72 h in the 10% glucose solution. After 7-8 days in the 10% solution, Saccharomyces cerevisiae produced ethanol at 100% of theoretical yield (5.0 g ethanol/100g solution), with a gradual decrease in alcohol production by 14 days. Immobilized Saccharomyces uvarum produced a maximum of 4.0 g ethanol/100 g solution within 2 days and then declined to ca. 1.0 g ethanol/100g solution after 7 days continuous fermentation in the 10% glucose solution. Zymomonas mobilis reached its maximum ethanol production at 4 days (4.7 g/100 g solution), and then diminished similarly to Saccharomyces uvarum. The development of a multiple disk shaft eliminated the problem both of uneven distribution of alginate-encapsulated cells and of glucose channeling within the continuous-flow fermentor column. This invention improved alcohol production about threefold for the yeast cells. (Refs. 13).

McGhee, J.E.; St. Julian, G.; Detroy, R.W.; Bothast, R.J.



[Mitochondria inheritance in yeast saccharomyces cerevisiae].  


The review is devoted to the main mechanisms of mitochondria inheritance in yeast Saccharonmyces cerevisiae. The genetic mechanisms of functionally active mitochondria inheritance in eukaryotic cells is one of the most relevant in modem researches. A great number of genetic diseases are associated with mitochondria dysfunction. Plasticity of eukaryotic cell metabolism according to the environmental changes is ensured by adequate mitochondria functioning by means of ATP synthesis coordination, reactive oxygen species accumulation, apoptosis regulation and is an important factor of cell adaptation to stress. Mitochondria participation in important for cell vitality processes masters the presence of accurate mechanisms of mitochondria functions regulation according to environment fluctuations. The mechanisms of mitochondria division and distribution are highly conserved. Baker yeast S. cerevisiae is an ideal model object for mitochondria researches due to energetic metabolism lability, ability to switch over respiration to fermentation, and petite-positive phenotype. Correction of metabolism according to the environmental changes is necessary for cell vitality. The influence of respiratory, carbon, amino acid and phosphate metabolism on mitochondria functions was shown. As far as the mechanisms that stabilize functions of mitochondria and mtDNA are highly conserve, we can project yeast regularities on higher eukaryotes systems. This makes it possible to approximate understanding the etiology and pathogenesis of a great number of human diseases. PMID:21786681

Fizikova, A Iu



Phosphoenolpyruvate Carboxykinase as the Sole Anaplerotic Enzyme in Saccharomyces cerevisiae?  

PubMed Central

Pyruvate carboxylase is the sole anaplerotic enzyme in glucose-grown cultures of wild-type Saccharomyces cerevisiae. Pyruvate carboxylase-negative (Pyc?) S. cerevisiae strains cannot grow on glucose unless media are supplemented with C4 compounds, such as aspartic acid. In several succinate-producing prokaryotes, phosphoenolpyruvate carboxykinase (PEPCK) fulfills this anaplerotic role. However, the S. cerevisiae PEPCK encoded by PCK1 is repressed by glucose and is considered to have a purely decarboxylating and gluconeogenic function. This study investigates whether and under which conditions PEPCK can replace the anaplerotic function of pyruvate carboxylase in S. cerevisiae. Pyc? S. cerevisiae strains constitutively overexpressing the PEPCK either from S. cerevisiae or from Actinobacillus succinogenes did not grow on glucose as the sole carbon source. However, evolutionary engineering yielded mutants able to grow on glucose as the sole carbon source at a maximum specific growth rate of ca. 0.14 h?1, one-half that of the (pyruvate carboxylase-positive) reference strain grown under the same conditions. Growth was dependent on high carbon dioxide concentrations, indicating that the reaction catalyzed by PEPCK operates near thermodynamic equilibrium. Analysis and reverse engineering of two independently evolved strains showed that single point mutations in pyruvate kinase, which competes with PEPCK for phosphoenolpyruvate, were sufficient to enable the use of PEPCK as the sole anaplerotic enzyme. The PEPCK reaction produces one ATP per carboxylation event, whereas the original route through pyruvate kinase and pyruvate carboxylase is ATP neutral. This increased ATP yield may prove crucial for engineering of efficient and low-cost anaerobic production of C4 dicarboxylic acids in S. cerevisiae.

Zelle, Rintze M.; Trueheart, Josh; Harrison, Jacob C.; Pronk, Jack T.; van Maris, Antonius J. A.



RAD50 protein of S.cerevisiae exhibits ATP-dependent DNA binding.  

PubMed Central

RAD50 function of Saccharomyces cerevisiae is required during vegetative growth for recombinational repair of DNA double strand breaks, and during meiosis for initiation of meiotic recombination and formation of synaptonemal complex. RAD50 encodes a 153 kDa polypeptide which includes an amino-terminal ATP binding domain essential for function and two long heptad repeat regions. We show below that RAD50 protein purified from yeast exhibits ATP-dependent binding to double stranded DNA. Physical properties of the purified protein are also described. Models for RAD50 function in vivo are discussed. Images

Raymond, W E; Kleckner, N



Frequency and Significance of Antibodies to Saccharomyces cerevisiae in Autoimmune Hepatitis  

Microsoft Academic Search

Our aims were to determine the frequency of antibodies to Saccharomyces cerevisiae in autoimmune hepatitis and assess associations with concurrent mucosal diseases, genetic factors, and corticosteroid response. Seropositivity was determined by enzyme immunoassay in 385 samples obtained from 178 patients. Antibodies to Saccharomyces cerevisiae were detected in 49 patients (28%), and serum levels of immunoglobulin A were higher in seropositive

Albert J. Czaja; Zakera Shums; Peter T. Donaldson; Gary L. Norman



D-arabinose dehydrogenase and its gene from Saccharomyces cerevisiae.  


D-Arabinose dehydrogenase was purified 843-fold from the cytosolic fraction of Saccharomyces cerevisiae with a recovery of 9%. The purified enzyme gave two bands with a molecular mass of 40 and 39 kDa on SDS-PAGE. The native enzyme had a molecular mass of 74 kDa as estimated by Sephacryl S-200 chromatography. Therefore, this enzyme was considered to be a heterodimer. The purified enzyme exhibited maximum activity at pH 10.0 and around 30 degrees C. The enzyme catalysed the oxidation of D-arabinose, L-xylose, L-fucose and L-galactose in the presence of NADP+. The apparent Km values at pH 10.0 with 50 microM NADP+ for D-arabinose, L-xylose, L-fucose, and L-galactose were 161, 24, 98 and 180 mM, respectively. The pH profile of Vmax and kcat/Km showed one ionisable groups around pH 8.3. D-Erythroascorbic acid was formed in vitro from D-arabinose by D-arabinose dehydrogenase and D-arabinono-1,4-lactone oxidase. The N-terminal amino acid sequence of the heavy subunit was Ser-Thr-Glu-Asn-Ile-Val-Glu-Asn-Met-Leu-His-Pro-Lys-Thr-. The N-terminus of the light subunit was blocked. The obtained peptide sequence was identical to the translational product of an unknown open reading frame, YBR149W, in chromosome II of S. cerevisiae. When compared with the translational product of this open reading frame, the peptide sequence was identical to the amino acid sequences of residues 7 to 20. The first six amino acids of this open reading frame were lost in protein sequence, which may be modified post-translationally. The heavy subunit was composed of 344 amino acid residues and its deduced amino acid sequence contained the motifs I, II, and III of aldo-keto reductase and also leucine zipper motif. This enzyme is the first heterodimeric protein of aldo-keto reductase family. In the deletion mutant of this gene, D-arabinose dehydrogenase activity and D-erythroascorbic acid were not detected. PMID:9920381

Kim, S T; Huh, W K; Lee, B H; Kang, S O



Functional characterization of human methylenetetrahydrofolate reductase in Saccharomyces cerevisiae.  


Human methylenetetrahydrofolate reductase (MTHFR, EC catalyzes the reduction of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate. 5-Methyltetrahydrofolate is a major methyl donor in the remethylation of homocysteine to methionine. Impaired MTHFR can cause high levels of homocysteine in plasma, which is an independent risk factor for vascular disease and neural tube defects. We have functionally characterized wild-type and several mutant alleles of human MTHFR in yeast, Saccharomyces cerevisiae. We have shown that yeast MET11 is a functional homologue of human MTHFR. Expression of the human MTHFR cDNA in a yeast strain deleted for MET11 can restore the strain's MTHFR activity in vitro and complement its methionine auxotrophic phenotype in vivo. To understand the domain structure of human MTHFR, we have truncated the C terminus (50%) of the protein and demonstrated that expressing an N-terminal human MTHFR in met11(-) yeast cells rescues the growth phenotype, indicating that this region contains the catalytic domain of the enzyme. However, the truncation leads to the reduced protein levels, suggesting that the C terminus may be important for protein stabilization. We have also functionally characterized four missense mutations identified from patients with severe MTHFR deficiency and two common missense polymorphisms found at high frequency in the general population. Three of the four missense mutations are unable to complement the auxotrophic phenotype of met11(-) yeast cells and show less than 7% enzyme activity of the wild type in vitro. Both of the two common polymorphisms are able to complement the growth phenotype, although one exhibited thermolabile enzyme activity in vitro. These results shall be useful for the functional characterization of MTHFR mutations and analysis structure/function relationship of the enzyme. PMID:10551815

Shan, X; Wang, L; Hoffmaster, R; Kruger, W D



Potassium is an activator of homoisocitrate dehydrogenase from Saccharomyces cerevisiae.  


Potassium is an activator of the reaction catalyzed by homoisocitrate (HIc) dehydrogenase (HIcDH) from Saccharomyces cerevisiae with either the natural substrate, homoisocitrate, or the slow substrate isocitrate. On the basis of initial velocity studies, the selectivity of the activator site for monovalent ions was determined. Potassium is the best activator, and NH 4 (+) and Rb (+) are also activators of the reaction, while Cs (+), Li (+), and Na (+) are not. Chloride inhibits the reaction, while acetate is much less effective. Substitution of potassium acetate for KCl changes the kinetic mechanism of HIcDH from a steady state random to a fully ordered mechanism with the binding of MgHIc followed by K (+) and NAD. The change in mechanism likely reflects an apparent increase in the affinity of enzyme for MgHIc as a result of elimination of the inhibitory effect of Cl (-). The V/K NAD pH-rate profile in the absence of K (+) exhibits a >10-fold decrease in the affinity of enzyme for NAD upon deprotonation of an enzyme side chain with a p K a of about 5.5-6. On the other hand, the affinity for NAD is relatively constant at high pH in the presence of 200 mM KCl. Since the affinity of the dinucleotide decreases as the enzyme group is protonated and the effect is overcome by a monovalent cation, the enzyme residue may be a neutral acid, aspartate or glutamate. Data suggest that K (+) replaces the proton, and likely binds to the enzyme residue, the pyrophosphoryl moiety of NAD, or both. Viscosity and solvent deuterium isotope effects studies suggest the isomerization of E-MgHIc binary complex limits the rate in the absence of K (+). PMID:18785753

Lin, Ying; West, Ann H; Cook, Paul F



Advanced biofuel production by the yeast Saccharomyces cerevisiae.  


Replacement of conventional transportation fuels with biofuels will require production of compounds that can cover the complete fuel spectrum, ranging from gasoline to kerosene. Advanced biofuels are expected to play an important role in replacing fossil fuels because they have improved properties compared with ethanol and some of these may have the energy density required for use in heavy duty vehicles, ships, and aviation. Moreover, advanced biofuels can be used as drop-in fuels in existing internal combustion engines. The yeast cell factory Saccharomyces cerevisiae can be turned into a producer of higher alcohols (1-butanol and isobutanol), sesquiterpenes (farnesene and bisabolene), and fatty acid ethyl esters (biodiesel), and here we discusses progress in metabolic engineering of S. cerevisiae for production of these advanced biofuels. PMID:23628723

Buijs, Nicolaas A; Siewers, Verena; Nielsen, Jens



Inactivation of Saccharomyces cerevisiae with radio frequency electric fields.  


The application of radio frequency (RF) electric fields as a nonthermal alternative to thermal inactivation of microorganisms in liquids was investigated. A novel RF system producing frequencies in the range of 20 to 60 kHz was developed. Electric field strengths of 20 and 30 kV/cm were applied to suspensions of Saccharomyces cerevisiae in water over a temperature range of 35 to 55 degrees C. The flow rate was 1.2 liters/min. The S. cerevisiae population was reduced by 2.1 +/- 0.1 log units following exposure to a 30-kV/cm field at 40 degrees C. The results of the present study provide the first evidence that strong RF electric fields inactivate microorganisms at moderately low temperatures. Increasing the field strength, the number of treatments, and the temperature enhanced inactivation. Frequency had no effect on inactivation over the range of frequencies studied. PMID:14503732

Geveke, David J; Brunkhorst, Christopher



Localization and targeting of isocitrate lyases in Saccharomyces cerevisiae.  

PubMed Central

Native isocitrate lyase from castor bean and a C-terminally truncated variant were expressed in Saccharomyces cerevisiae under the control of a galactose-inducible promoter. Both forms of isocitrate lyase were targeted to the yeast peroxisomes. They co-fractionated with catalase on sucrose-density-gradient centrifugation of a post-nuclear supernatant prepared from cells grown on oleic acid plus galactose, but were found in the cytosolic fractions when the cells were grown under conditions that repress peroxisome formation. The endogenous S. cerevisiae isocitrate lyase was found solely in the cytoplasmic fractions, even under growth conditions that induce peroxisome proliferation. This result shows that the presence of isocitrate lyase in peroxisomes is not essential for a functional glyoxylate cycle. Although the heterologous enzyme was transported to peroxisomes it was not enzymically active. Immunocytochemical studies provide independent evidence that the plant enzyme is imported into the matrix of yeast peroxisomes.

Taylor, K M; Kaplan, C P; Gao, X; Baker, A



Interorganelle signaling is a determinant of longevity in Saccharomyces cerevisiae.  

PubMed Central

Replicative capacity, which is the number of times an individual cell divides, is the measure of longevity in the yeast Saccharomyces cerevisiae. In this study, a process that involves signaling from the mitochondrion to the nucleus, called retrograde regulation, is shown to determine yeast longevity, and its induction resulted in postponed senescence. Activation of retrograde regulation, by genetic and environmental means, correlated with increased replicative capacity in four different S. cerevisiae strains. Deletion of a gene required for the retrograde response, RTG2, eliminated the increased replicative capacity. RAS2, a gene previously shown to influence longevity in yeast, interacts with retrograde regulation in setting yeast longevity. The molecular mechanism of aging elucidated here parallels the results of genetic studies of aging in nematodes and fruit flies, as well as the caloric restriction paradigm in mammals, and it underscores the importance of metabolic regulation in aging, suggesting a general applicability.

Kirchman, P A; Kim, S; Lai, C Y; Jazwinski, S M



Quantifying the complexities of Saccharomyces cerevisiae's ecosystem engineering via fermentation.  


The theory of niche construction suggests that organisms may engineer environments via their activities. Despite the potential of this phenomenon being realized by Darwin, the capability of niche construction to generally unite ecological and evolutionary biology has never been empirically quantified. Here I quantify the fitness effects of Saccharomyces cerevisiae's ecosystem engineering in a natural ferment in order to understand the interaction between ecological and evolutionary processes. I show that S. cerevisiae eventually dominates in fruit niches, where it is naturally initially rare, by modifying the environment through fermentation (the Crabtree effect) in ways which extend beyond just considering ethanol production. These data show that an additional cause of S. cerevisiae's competitive advantage over the other yeasts in the community is due to the production of heat via fermentation. Even though fermentation is less energetically efficient than respiration, it seems that this trait has been selected for because its net effect provides roughly a 7% fitness advantage over the other members of the community. These data provide an elegant example of niche construction because this trait clearly modifies the environment and therefore the selection pressures to which S. cerevisiae, and other organisms that access the fruit resource, including humans, are exposed to. PMID:18724717

Goddard, Matthew R



Global analysis of RNA oxidation in Saccharomyces cerevisiae  

PubMed Central

Oxidative RNA damage has been linked to loss of RNA function and to the development of many human age-related diseases. Consequently, a need exists for methods to identify and quantify the extent of RNA oxidation on a genome-wide basis. We developed such a method by combining affinity selection of mRNA containing 8-hydroxyguanine with high throughput DNA sequencing. We demonstrate that this assay is suitable for detecting differences in the extent of oxidation between RNA transcripts. We applied this method to the yeast Saccharomyces cerevisiae grown under physiological conditions and in response to hydrogen peroxide, and detected significantly oxidized RNA transcripts.

McKinlay, Anastasia; Gerard, Wayne; Fields, Stanley



Homeostatic and Adaptive Responses to Zinc Deficiency in Saccharomyces cerevisiae*  

PubMed Central

Zinc is an essential nutrient and serves as a structural or catalytic cofactor for many proteins. Thus, cells need mechanisms to maintain zinc homeostasis when available zinc supplies decrease. In addition, cells require other mechanisms to adapt intracellular processes to suboptimal levels of zinc. By exploring the transcriptional responses to zinc deficiency, studies of the yeast Saccharomyces cerevisiae have revealed both homeostatic and adaptive responses to low zinc. The Zap1 zinc-responsive transcription factor regulates several genes in yeast, and the identity of these genes has led to new insights regarding how cells respond to the stress of zinc deficiency.

Eide, David J.



The nucleotide sequence of Saccharomyces cerevisiae chromosome V  

PubMed Central

Here we report the sequence of 569,202 base pairs of Saccharomyces cerevisiae chromosome V. Analysis of the sequence revealed a centromere, two telomeres and 271 open reading frames (ORFs) plus 13 tRNAs and four small nuclear RNAs. There are two Ty1 transposable elements, each of which contains an ORF (included in the count of 271). Of the ORFs, 78 (29%) are new, 81 (30%) have potential homologues in the public databases, and 112 (41%) are previously characterized yeast genes.

Dietrich, F. S.; Mulligan, J.; Hennessy, K.; Yelton, M. A.; Allen, E.; Araujo, R.; Aviles, E.; Berno, A.; Brennan, T.; Carpenter, J.; Chen, E.; Cherry, J. M.; Chung, E.; Duncan, M.; Guzman, E.; Hartzell, G.; Hunicke-Smith, S.; Hyman, R. W.; Kayser, A.; Komp, C.; Lashkari, D.; Lew, H.; Lin, D.; Mosedale, D.; Nakahara, K.; Namath, A.; Norgren, R.; Oefner, P.; Oh, C.; Petel, F. X.; Roberts, D.; Sehl, P.; Schramm, S.; Shogren, T.; Smith, V.; Taylor, P.; Wei, Y.; Botstein, D.; Davis, R. W.



Isobutanol production from D-xylose by recombinant Saccharomyces cerevisiae.  


Simultaneous overexpression of an optimized, cytosolically localized valine biosynthesis pathway together with overexpression of xylose isomerase XylA from Clostridium phytofermentans, transaldolase Tal1 and xylulokinase Xks1 enabled recombinant Saccharomyces cerevisiae cells to complement the valine auxotrophy of ilv2,3,5 triple deletion mutants for growth on D-xylose as the sole carbon source. Moreover, after additional overexpression of ketoacid decarboxylase Aro10 and alcohol dehydrogenase Adh2, the cells were able to ferment D-xylose directly to isobutanol. PMID:23279585

Brat, Dawid; Boles, Eckhard



DNA sequence analysis of spontaneous mutagenesis in Saccharomyces cerevisiae.  

PubMed Central

To help elucidate the mechanisms involved in spontaneous mutagenesis, DNA sequencing has been applied to characterize the types of mutation whose rates are increased or decreased in mutator or antimutator strains, respectively. Increased spontaneous mutation rates point to malfunctions in genes that normally act to reduce spontaneous mutation, whereas decreased rates are associated with defects in genes whose products are necessary for spontaneous mutagenesis. In this article, we survey and discuss the mutational specificities conferred by mutator and antimutator genes in the budding yeast Saccharomyces cerevisiae. The implications of selected aspects of the data are considered with respect to the mechanisms of spontaneous mutagenesis.

Kunz, B A; Ramachandran, K; Vonarx, E J



Stable denaturation of chromosomal DNA from Saccharomyces cerevisiae during meiosis.  

PubMed Central

Partial denaturation of Saccharomyces cerevisiae chromosomal DNA was found to occur spontaneously during meiosis. Short regions of strand separation (300 base pairs long) were seen in DNA molecules prepared for electron microscopy by the aqueous spreading technique. These regions were clustered along the DNA. The time course of their appearance indicated that the denatured regions were present during the periods of premeiotic DNA replication and recombination. A similar pattern of denaturation was also detected in the DNA from vegetatively grown cells of a conditional cdc8 mutant, which is defective in DNA replication. Images

Klein, H L; Byers, B



Nutritional Control via Tor signaling in Saccharomyces cerevisiae  

PubMed Central

The yeast Saccharomyces cerevisiae senses and responds to nutrients by adapting its growth rate and undergoing morphogenic transitions to ensure survival. The Tor pathway is a major integrator of nutrient-derived signals that in coordination with other signaling pathways orchestrates cell growth. Recent advances have identified novel Tor kinase substrates and established the protein trafficking membranous network and the nucleus as platforms for Tor signaling. These and other recent findings delineate distinct signaling branches emanating from membrane associated Tor complexes to control cell growth.

Rohde, John R.; Bastidas, Robert; Puria, Rekha; Cardenas, Maria E.



SPME-GC method as a tool to differentiate VOC profiles in Saccharomyces cerevisiae wine yeasts.  


The aim of this work was to study the variability of 36 Saccharomyces cerevisiae wild strains isolated from different grape varieties and from two very distant zones, located in Northern and Southern Italy. The strains were differentiated on the basis of parameters of technological interest, such as resistance to antimicrobial compounds frequently present in wine, and the production of volatile aromatic compounds (VOC), determined by SPME procedure in the experimental wines obtained by inoculated fermentations. The VOC profile allowed to differentiate the yeasts in function of isolation area: S. cerevisiae isolated from Southern Italy grapes were able to produce more volatile compounds than those from Northern Italy. The compounds synthesized by all the yeasts, besides the ethanol, were 3-methyl-1-butanol and ethyl acetate. The production of acids during the alcoholic fermentation was a characteristic of Southern yeast strains. The screening of S. cerevisiae strains for technological parameters, such as sulphur dioxide, copper and ethanol resistance or hydrogen sulphide production, revealed similar behaviour for sulphur dioxide resistance among Northern and Southern S. cerevisiae strains. Copper resistance and sulphur dioxide production were correlated to isolation area: S. cerevisiae "Northern" strains showed higher copper resistance and lowest hydrogen sulphide production than that exhibited from "Southern" strains. PMID:19269564

Mauriello, Giacomo; Capece, Angela; D'Auria, Maurizio; Garde-Cerdán, Teresa; Romano, Patrizia



Asparaginase II of Saccharomyces cerevisiae. Characterization of the ASP3 gene.  


Purified preparations of asparaginase II of Saccharomyces cerevisiae exhibit two protein bands upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cloning and sequencing of the ASP3 gene, and partial amino acid sequencing as asparaginase II, imply that both bands are encoded by ASP3 but have different N termini. Northern blot analysis using the cloned ASP3 gene as a probe indicates that nitrogen catabolite repression of asparaginase II is achieved by alteration in mRNA levels. Deletion of sequences greater than 600 base pairs upstream from the initiation AUG codon results in an altered response to certain nitrogen sources in strains containing the truncated gene. PMID:3042786

Kim, K W; Kamerud, J Q; Livingston, D M; Roon, R J



Biosynthesis of Riboflavine in Saccharomyces cerevisiae: the Role of Genes rib1 and rib7  

PubMed Central

Haploid strains of Saccharomyces cerevisiae with mutations in two different rib genes were constructed. These strains were studied by tetrad analysis and by quantitative determination of accumulation products. The genes rib1, rib7, and rib2 are not linked to each other. rib1-rib7 strains and rib1-rib2 strains exhibit the phenotypic properties of rib1 strains. rib7-rib2 strains show the phenotypic properties of rib7 strains. The results support the conclusion that the genes rib1 and rib7 code for the first and second enzyme of the riboflavine pathway, respectively.

Oltmanns, O.; Bacher, A.



Ureidosuccinate is transported by the allantoate transport system in Saccharomyces cerevisiae.  

PubMed Central

The regulatory characteristics exhibited by ureidosuccinate transport in Saccharomyces cerevisiae led us to hypothesize that this biosynthetic intermediate was transported via the degradative allantoate transport system. The hypothesis was verified by the finding that neither dal5 nor urep1 mutant strains could transport allantoate or ureidosuccinate. Mutations in the two loci were tightly linked and failed to complement one another, suggesting that they were allelic. The use of a common transport system for accumulation of both biosynthetic and degradative metabolites explains the paradoxical characteristics observed for control of ureidosuccinate and allantoate transport.

Turoscy, V; Cooper, T G



Functional Expression of a Bacterial Xylose Isomerase in Saccharomyces cerevisiae?  

PubMed Central

In industrial fermentation processes, the yeast Saccharomyces cerevisiae is commonly used for ethanol production. However, it lacks the ability to ferment pentose sugars like d-xylose and l-arabinose. Heterologous expression of a xylose isomerase (XI) would enable yeast cells to metabolize xylose. However, many attempts to express a prokaryotic XI with high activity in S. cerevisiae have failed so far. We have screened nucleic acid databases for sequences encoding putative XIs and finally were able to clone and successfully express a highly active new kind of XI from the anaerobic bacterium Clostridium phytofermentans in S. cerevisiae. Heterologous expression of this enzyme confers on the yeast cells the ability to metabolize d-xylose and to use it as the sole carbon and energy source. The new enzyme has low sequence similarities to the XIs from Piromyces sp. strain E2 and Thermus thermophilus, which were the only two XIs previously functionally expressed in S. cerevisiae. The activity and kinetic parameters of the new enzyme are comparable to those of the Piromyces XI. Importantly, the new enzyme is far less inhibited by xylitol, which accrues as a side product during xylose fermentation. Furthermore, expression of the gene could be improved by adapting its codon usage to that of the highly expressed glycolytic genes of S. cerevisiae. Expression of the bacterial XI in an industrially employed yeast strain enabled it to grow on xylose and to ferment xylose to ethanol. Thus, our findings provide an excellent starting point for further improvement of xylose fermentation in industrial yeast strains.

Brat, Dawid; Boles, Eckhard; Wiedemann, Beate



Propionate metabolism in Saccharomyces cerevisiae: implications for the metabolon hypothesis.  


Aerobic, glucose-limited chemostat of Saccharomyces cerevisiae CBS 8066 co-metabolized propionate when this compound was added to the reservoir medium. Co-metabolism of propionate led to an increase of the biomass and protein yields. Attempts to grow S. cerevisiae on propionate as a sole source of carbon and energy were not successful. Activities of propionyl-CoA synthetase in cell-free extracts were sufficient to account for the rates of propionate consumption observed in the chemostat cultures. Activities of propionyl-CoA carboxylase, a key enzyme of the methylmalonyl-CoA pathway of propionate metabolism, were negligible. In contrast, activities of 2-methylcitrate synthase, a key enzyme activity of the 2-methylcitrate pathway of propionate metabolism, increased substantially with increasing propionate-to-glucose ratios in the reservoir media, and were sufficient to account for the propionate consumption rates observed in the chemostat cultures. This suggested that the 2-methylcitrate pathway is the major pathway of propionate metabolism in S. cerevisiae. In the literature, labelling patterns observed after incubation of this yeast with [3-13C]propionate have been interpreted as evidence for channelling of tricarboxylic acid (TCA) cycle intermediates, possibly as a consequence of the organization of TCA cycle enzymes in a metabolon. However, this interpretation of 13C-labelling patterns rested on the assumption that propionate metabolism in S. cerevisiae occurs via the methylmalonyl-CoA pathway. Since the distribution of 13C in alanine reported in the literature is fully compatible with a major role of the 2-methylcitrate pathway in propionate metabolism, it cannot be interpreted as evidence for the existence of a TCA cycle metabolon in S. cerevisiae. PMID:7912143

Pronk, J T; van der Linden-Beuman, A; Verduyn, C; Scheffers, W A; van Dijken, J P



Microcalorimetric monitoring of growth of Saccharomyces cerevisiae: osmotolerance in relation to physiological state.  

PubMed Central

The importance of the physiological state of a culture of Saccharomyces cerevisiae for tolerance to sudden osmotic dehydration was studied, and it was investigated whether specific osmotolerance factors were demonstrable. The microcalorimeter was used to monitor growth, and different physiological states of the culture were selected and their osmotolerance was tested. In addition to cells in the stationary phase, cells from the transition phase between respirofermentative and respiratory catabolism were osmotolerant. S. cerevisiae exhibited ever-changing metabolism during batch growth on either glucose or ethanol as the carbon source. Instantaneous heat production per biomass formation (dQ/dX) and specific activity of sn-glycerol 3-phosphate dehydrogenase (GPDH) (EC were shown to differ for different physiological states. Neither high respiratory activity nor low total cellular activity, nor factors involved in osmoregulation, i.e., intracellular glycerol or activity of GPDH, correlated with the osmotolerant phenotype.

Blomberg, A; Larsson, C; Gustafsson, L



Variations of two pools of glycogen and carbohydrate in Saccharomyces cerevisiae grown with various ethanol concentrations.  


Glycogen, a major reservoir of energy in Saccharomyces cerevisiae, is found to be present as soluble and membrane-bound insoluble pools. Yeast cells can store excess glycogen when grown in media with higher concentration of sugar or when subjected to nutritional stress conditions. Saccharomyces cerevisiae NCIM-3300 was grown in media having ethanol concentrations up to 12% (v/v). The effects of externally added ethanol on glycogen and other carbohydrate content of yeast were studied by using alkali digestion process. Fermentative activities of cells grown in the presence of various ethanol concentrations (2-8% v/v) exhibited increase in values of glycogen and other carbohydrate, whereas cells grown with higher concentrations of ethanol (10-12% v/v) exhibited depletion in glycogen and carbohydrate content along with decrease in cell weight. Such inhibitory effect of ethanol was also exhibited in terms of reduction in total cell count of yeast grown in media with 2-16% (v/v) ethanol and 8% (w/v) sugar. These data suggest that, as the plasma membrane is a prime target for ethanol action, membrane-bound insoluble glycogen might play a protective role in combating ethanol stress. Elevated level of cell-surface alpha-glucans in yeast grown with ethanol, as measured by using amyloglucosidase treatment, confirms the correlation between ethanol and glycogen. PMID:20373126

Dake, M S; Jadhv, J P; Patil, N B



Transformations of inorganic mercury by Candida albicans and Saccharomyces cerevisiae  

SciTech Connect

Saccharomyces cerevisiae and Candida albicans were incubated with 0.25, 0.5, or 0.75 {mu}g of Hg (as HgCl{sub 2}) per ml of Nelson's medium in the presence of trace amounts of oxygen at 28{degree}C for 12 days. Two control media were used, one without added Hg and one without yeast inoculum. Yeast cell growth was estimated after 1, 2, 3, and 8 days of incubation. The contents of organomercury in the system and of elemental mercury released from the media and collected in traps were determined at the end of the experiments. The results were as follows: (1) C. albicans was the more mercury-resistant species, but both yeast species failed to grown in the media containing 0.75 {mu}g of Hg per ml.; (2) The amounts of organomercury produced by the two species were proportional to the amount of HgCl{sub 2} added to the medium. In all cases C. albicans produced considerably larger amounts of methylmercury than S. cerevisiae; (3) The amounts of elemental Hg produced were inversely proportional to the HgCl{sub 2} level added in the case of S. cerevisiae but were all similar in the case of C. albicans;and (4) Neither organomercury nor elemental Hg was produced in any of the control media.

Yannai, S.; Berdicevsky, I.; Duek, L. (Technion-Israel Institute of Technology, Haifa (Israel))



Biogeographical characterization of Saccharomyces cerevisiae wine yeast by molecular methods.  


Biogeography is the descriptive and explanatory study of spatial patterns and processes involved in the distribution of biodiversity. Without biogeography, it would be difficult to study the diversity of microorganisms because there would be no way to visualize patterns in variation. Saccharomyces cerevisiae, "the wine yeast," is the most important species involved in alcoholic fermentation, and in vineyard ecosystems, it follows the principle of "everything is everywhere." Agricultural practices such as farming (organic versus conventional) and floor management systems have selected different populations within this species that are phylogenetically distinct. In fact, recent ecological and geographic studies highlighted that unique strains are associated with particular grape varieties in specific geographical locations. These studies also highlighted that significant diversity and regional character, or 'terroir,' have been introduced into the winemaking process via this association. This diversity of wild strains preserves typicity, the high quality, and the unique flavor of wines. Recently, different molecular methods were developed to study population dynamics of S. cerevisiae strains in both vineyards and wineries. In this review, we will provide an update on the current molecular methods used to reveal the geographical distribution of S. cerevisiae wine yeast. PMID:23805132

Tofalo, Rosanna; Perpetuini, Giorgia; Schirone, Maria; Fasoli, Giuseppe; Aguzzi, Irene; Corsetti, Aldo; Suzzi, Giovanna



Osmo-, Thermo- and Ethanol- Tolerances of Saccharomyces cerevisiae S1  

PubMed Central

Saccharomyces cerevisiae S1, which is a locally isolated and improved strain showed viability at 40, 45 and 50°C and produced ethanol at 40, 43 and 45°C. When the cells were given heat shock at 45°C for 30min and grown at 40°C, 100% viability was observed for 60h, and addition of 200gL?1 ethanol has led to complete cell death at 30h. Heat shock given at 45°C (for 30min) has improved the tolerance to temperature induced ethanol shock leading to 37% viability at 30h. When the cells were subjected to ethanol (200gL?1 for 30 min) and osmotic shock (sorbitol 300gL?1), trehalose contents in the cells were increased. The heat shocked cells showed better viability in presence of added ethanol. Soy flour supplementation has improved the viability of S. cerevisiae S1 to 80% in presence of 100gL?1 added ethanol and to 60% in presence of 300gL?1sorbitol. In presence of sorbitol (200gL?1) and ethanol (50gL?1) at 40°C, 46% viability was retained by S. cerevisiae S1 at 48h and it was improved to 80% by soy flour supplementation.

Balakumar, Sandrasegarampillai; Arasaratnam, Vasanthy



Nature and distribution of large sequence polymorphisms in Saccharomyces cerevisiae  

PubMed Central

To obtain a better understanding of the genome-wide distribution and the nature of large sequence polymorphisms (LSPs) in Saccharomyces cerevisiae, we hybridized genomic DNA of 88 haploid or homozygous diploid S. cerevisiae strains of diverse geographic origins and source substrates onto high-density tiling arrays. Based on loss of hybridization, we identified 384 LSPs larger than 500 bp that were located in 188 non-overlapping regions of the genome. Validation by polymerase chain reaction-amplification and/or DNA sequencing revealed that 39 LSPs were due to deletions, while 74 LSPs involved sequences diverged far enough from the S288c reference genome sequence as to prevent hybridization to the microarray features. The LSP locations were biased towards the subtelomeric regions of chromosomes, where high genetic variation in genes involved in transport or fermentation is thought to facilitate rapid adaptation of S. cerevisiae to new environments. The diverged LSP sequences appear to have different allelic ancestries and were in many cases identified as S. paradoxus introgressions.

Muller, Ludo A. H.; McCusker, John H.



Biogeographical characterization of Saccharomyces cerevisiae wine yeast by molecular methods  

PubMed Central

Biogeography is the descriptive and explanatory study of spatial patterns and processes involved in the distribution of biodiversity. Without biogeography, it would be difficult to study the diversity of microorganisms because there would be no way to visualize patterns in variation. Saccharomyces cerevisiae, “the wine yeast,” is the most important species involved in alcoholic fermentation, and in vineyard ecosystems, it follows the principle of “everything is everywhere.” Agricultural practices such as farming (organic versus conventional) and floor management systems have selected different populations within this species that are phylogenetically distinct. In fact, recent ecological and geographic studies highlighted that unique strains are associated with particular grape varieties in specific geographical locations. These studies also highlighted that significant diversity and regional character, or ‘terroir,’ have been introduced into the winemaking process via this association. This diversity of wild strains preserves typicity, the high quality, and the unique flavor of wines. Recently, different molecular methods were developed to study population dynamics of S. cerevisiae strains in both vineyards and wineries. In this review, we will provide an update on the current molecular methods used to reveal the geographical distribution of S. cerevisiae wine yeast.

Tofalo, Rosanna; Perpetuini, Giorgia; Schirone, Maria; Fasoli, Giuseppe; Aguzzi, Irene; Corsetti, Aldo; Suzzi, Giovanna



Production of aromatics in Saccharomyces cerevisiae--a feasibility study.  


Aromatics are amongst the most important bulk feedstocks for the chemical industry, however, no viable bioprocess exists today and production is still dependent on petro-chemistry. In this article the production of aromatic precursors such as p-hydroxybenzoic acid (PHBA) and p-amino benzoic acid (PABA) in Saccharomyces cerevisiae was evaluated using metabolic network analysis. Theoretical mass yields for PHBA and for PABA obtained by metabolic network analysis were 0.58 and 0.53 g g(glucose)?¹, respectively. A major setback for microbial production of aromatics is the high toxicity of the products. Therefore, PHBA and PABA toxicity was evaluated in S. cerevisiae. Minimal inhibitory concentrations of 38.3 g L?¹ for PHBA and 0.62 g L?¹ for PABA were observed. However, PABA toxicity could be alleviated in adaptation experiments. Finally, metabolic engineering was used to create proof of principle first generation strains of S. cerevisiae. Overall accumulation of 650 ?M PHBA and 250 ?M PABA could be achieved. PMID:22579724

Krömer, Jens O; Nunez-Bernal, Dariela; Averesch, Nils J H; Hampe, Jennifer; Varela, Javier; Varela, Cristian



Indirect immunofluorescence labeling in the yeast Saccharomyces cerevisiae.  


The budding yeast Saccharomyces cerevisiae is an ideal model system for the initial characterization of novel genes and their corresponding gene products. Genetic analysis is straightforward, and sophisticated cell biological methods are available. Furthermore, only in a genetically tractable organism is it possible to use epitope-tagged proteins to the best advantage. In S. cerevisiae, it is a trivial matter to determine whether a modified version of a protein is functional. This is accomplished by testing whether the tagged protein can complement a null phenotype or rescue a conditional phenotype. Methods such as indirect immunofluorescence and the use of green fluorescent protein (GFP) allow the investigator to correlate the intracellular localization of the protein with its function in vivo. This article includes a detailed protocol for performing indirect immunofluorescence with S. cerevisiae. Cells are grown exponentially and then fixed. After fixation, an enzyme is used to remove the cell wall, and the cells are adhered to slides. Subsequent steps involve the application of primary and secondary antibodies and the final processing of the slide. PMID:20150056

Silver, Pamela



The postmitotic Saccharomyces cerevisiae after spaceflight showed higher viability  

NASA Astrophysics Data System (ADS)

The budding yeast Saccharomyces cerevisiae has been proposed as an ideal model organism for clarifying the biological effects caused by spaceflight conditions. The postmitotic S. cerevisiae cells onboard Practice eight recoverable satellite were subjected to spaceflight for 15 days. After recovery, the viability, the glycogen content, the activities of carbohydrate metabolism enzymes, the DNA content and the lipid peroxidation level in yeast cells were analyzed. The viability of the postmitotic yeast cells after spaceflight showed a three-fold increase as compared with that of the ground control cells. Compared to the ground control cells, the lipid peroxidation level in the spaceflight yeast cells markedly decreased. The spaceflight yeast cells also showed an increase in G2/M cell population and a decrease in Sub-G1 cell population. The glycogen content and the activities of hexokinase and succinate dehydrogenase significantly decreased in the yeast cells after spaceflight. In contrast, the activity of malate dehydrogenase showed an obvious increase after spaceflight. These results suggested that microgravity or spaceflight could promote the survival of postmitotic S. cerevisiae cells through regulating carbohydrate metabolism, ROS level and cell cycle progression.

Yi, Zong-Chun; Li, Xiao-Fei; Wang, Yan; Wang, Jie; Sun, Yan; Zhuang, Feng-Yuan



Expression of Pneumocystis jirovecii major surface glycoprotein in Saccharomyces cerevisiae.  


The major surface glycoprotein (Msg), which is the most abundant protein expressed on the cell surface of Pneumocystis organisms, plays an important role in the attachment of this organism to epithelial cells and macrophages. In the present study, we expressed Pneumocystis jirovecii Msg in Saccharomyces cerevisiae, a phylogenetically related organism. Full-length P. jirovecii Msg was expressed with a DNA construct that used codons optimized for expression in yeast. Unlike in Pneumocystis organisms, recombinant Msg localized to the plasma membrane of yeast rather than to the cell wall. Msg expression was targeted to the yeast cell wall by replacing its signal peptide, serine-threonine-rich region, and glycophosphatidylinositol anchor signal region with the signal peptide of cell wall protein ?-agglutinin of S. cerevisiae, the serine-threonine-rich region of epithelial adhesin (Epa1) of Candida glabrata, and the carboxyl region of the cell wall protein (Cwp2) of S. cerevisiae, respectively. Immunofluorescence analysis and treatment with ?-1,3 glucanase demonstrated that the expressed Msg fusion protein localized to the yeast cell wall. Surface expression of Msg protein resulted in increased adherence of yeast to A549 alveolar epithelial cells. Heterologous expression of Msg in yeast will facilitate studies of the biologic properties of Pneumocystis Msg. PMID:23532098

Kutty, Geetha; England, Katherine J; Kovacs, Joseph A



Ciclohexadespipeptide beauvericin degradation by different strains of Saccharomyces cerevisiae.  


The interaction between the mycotoxin beauvericin (BEA) and 9 yeast strains of Saccharomyces cerevisiae named LO9, YE-2, YE5, YE-6, YE-4, A34, A17, A42 and A08 was studied. The biological degradations were carried out under aerobic conditions in the liquid medium of Potato Dextrose Broth (PDB) at 25°C for 48h and in a food/feed system composed of corn flour at 37°C for 3days, respectively. BEA present in fermented medium and corn flour was determined using liquid chromatography coupled to the mass spectrometry detector in tandem (LC-MS/MS) and the BEA degradation products produced during the fermentations were determined using the technique of the liquid chromatography coupled to a linear ion trap (LIT). Results showed that the S. cerevisiae strains reduced meanly the concentration of the BEA present in PDB by 86.2% and in a food system by 71.1%. All the S. cerevisiae strains used in this study showed a significant BEA reduction during the fermentation process employed. PMID:23791659

Meca, G; Zhou, T; Li, X Z; Ritieni, A; Mañes, J



PGM2 overexpression improves anaerobic galactose fermentation in Saccharomyces cerevisiae  

PubMed Central

Background In Saccharomyces cerevisiae galactose is initially metabolized through the Leloir pathway after which glucose 6-phosphate enters glycolysis. Galactose is controlled both by glucose repression and by galactose induction. The gene PGM2 encodes the last enzyme of the Leloir pathway, phosphoglucomutase 2 (Pgm2p), which catalyses the reversible conversion of glucose 1-phosphate to glucose 6-phosphate. Overexpression of PGM2 has previously been shown to enhance aerobic growth of S. cerevisiae in galactose medium. Results In the present study we show that overexpression of PGM2 under control of the HXT7'promoter from an integrative plasmid increased the PGM activity 5 to 6 times, which significantly reduced the lag phase of glucose-pregrown cells in an anaerobic galactose culture. PGM2 overexpression also increased the anaerobic specific growth rate whereas ethanol production was less influenced. When PGM2 was overexpressed from a multicopy plasmid instead, the PGM activity increased almost 32 times. However, this increase of PGM activity did not further improve aerobic galactose fermentation as compared to the strain carrying PGM2 on the integrative plasmid. Conclusion PGM2 overexpression in S. cerevisiae from an integrative plasmid is sufficient to reduce the lag phase and to enhance the growth rate in anaerobic galactose fermentation, which results in an overall decrease in fermentation duration. This observation is of particular importance for the future development of stable industrial strains with enhanced PGM activity.



Antimicrobial action of palmarosa oil (Cymbopogon martinii) on Saccharomyces cerevisiae.  


The essential oil extracted from palmarosa (Cymbopogon martinii) has proven anti-microbial properties against cells of Saccharomyces cerevisiae. Low concentrations of the oil (0.1%) inhibited the growth of S. cerevisiae cells completely. The composition of the sample of palmarosa oil was determined as 65% geraniol and 20% geranyl acetate as confirmed by GC-FTIR. The effect of palmarosa oil in causing K(+) leakage from yeast cells was attributed mainly to geraniol. Some leakage of magnesium ions was also observed. Blocking potassium membrane channels with caesium ions before addition of palmarosa oil did not change the extent of K(+) ion leakage, which was equal to the total sequestered K(+) in the cells. Palmarosa oil led to changes in the composition of the yeast cell membrane, with more saturated and less unsaturated fatty acids in the membrane after exposure of S. cerevisiae cells to the oil. Some of the palmarosa oil was lost by volatilization during incubation of the oil with the yeast cells. The actual concentration of the oil components affecting the yeast cells could not therefore be accurately determined. PMID:12809717

Prashar, Anjali; Hili, Pauline; Veness, Robert G; Evans, Christine S



Replicative Deoxyribonucleic Acid Synthesis in Isolated Mitochondria from Saccharomyces cerevisiae  

PubMed Central

The characteristics of a system for the in vitro synthesis of mitochondrial deoxyribonucleic acid (mtDNA) in mitochondria isolated from Saccharomyces cerevisiae are described. In this system the exclusive product of the reaction is mtDNA. Under optimal conditions the initial rate of synthesis is close to the calculated in vivo rate; the rate is approximately linear for 20 min but then decreases gradually with time. DNA synthesis proceeds for at least 60 min and the de novo synthesis of an amount of mtDNA equivalent to 15% of the mtDNA initially present is achieved. The rate and extent of synthesis observed with mitochondria isolated from grande and petite (rho?) strains were similar. The mode of DNA synthesis is semiconservative; after density labeling with 5-bromodeoxyuridine triphosphate, in vitro, the majority of labeled DNA fragments of duplex molecular weight, 6 × 106, are of a density close to that calculated for hybrid yeast mtDNA. The density label is incorporated into one strand of the duplex molecules. These properties indicate that the synthesis resembles replicative rather than repair synthesis. This system therefore provides a convenient method for the study of mtDNA synthesis in S. cerevisiae. The observation that mtDNA synthesis is semiconservative in vitro suggests that the dispersive mode of synthesis observed in S. cerevisiae in vivo labeling studies is the result of some other process, possibly a high recombination rate.

Mattick, John S.; Hall, Ruth M.



Overall kinetic mechanism of saccharopine dehydrogenase from Saccharomyces cerevisiae.  


Kinetic data have been measured for the histidine-tagged saccharopine dehydrogenase from Saccharomyces cerevisiae, suggesting the ordered addition of nicotinamide adenine dinucleotide (NAD) followed by saccharopine in the physiologic reaction direction. In the opposite direction, the reduced nicotinamide adenine dinucleotide (NADH) adds to the enzyme first, while there is no preference for the order of binding of alpha-ketoglutarate (alpha-Kg) and lysine. In the direction of saccharopine formation, data also suggest that, at high concentrations, lysine inhibits the reaction by binding to free enzyme. In addition, uncompetitive substrate inhibition by alpha-Kg and double inhibition by NAD and alpha-Kg suggest the existence of an abortive E:NAD:alpha-Kg complex. Product inhibition by saccharopine is uncompetitive versus NADH, suggesting a practical irreversibility of the reaction at pH 7.0 in agreement with the overall K(eq). Saccharopine is noncompetitive versus lysine or alpha-Kg, suggesting the existence of both E:NADH:saccharopine and E:NAD:saccharopine complexes. NAD is competitive versus NADH, and noncompetitive versus lysine and alpha-Kg, indicating the combination of the dinucleotides with free enzyme. Dead-end inhibition studies are also consistent with the random addition of alpha-Kg and lysine. Leucine and oxalylglycine serve as lysine and alpha-Kg dead-end analogues, respectively, and are uncompetitive against NADH and noncompetitive against alpha-Kg and lysine, respectively. Oxaloacetate (OAA), pyruvate, and glutarate behave as dead-end analogues of lysine, which suggests that the lysine-binding site has a higher affinity for keto acid analogues than does the alpha-Kg site or that dicarboxylic acids have more than one binding mode on the enzyme. In addition, OAA and glutarate also bind to free enzyme as does lysine at high concentrations. Glutarate gives S-parabolic noncompetitive inhibition versus NADH, indicating the formation of a E:(glutarate)2 complex as a result of occupying both the lysine- and alpha-Kg-binding sites. Pyruvate, a slow alternative keto acid substrate, exhibits competitive inhibition versus both lysine and alpha-Kg, suggesting the combination to the E:NADH:alpha-Kg and E:NADH:lysine enzyme forms. The equilibrium constant for the reaction has been measured at pH 7.0 as 3.9 x 10(-7) M by monitoring the change in NADH upon the addition of the enzyme. The Haldane relationship is in very good agreement with the directly measured value. PMID:17002315

Xu, Hengyu; West, Ann H; Cook, Paul F



Preeclamptic cord blood hemolysis and the effect of Monascus purpureus and Saccharomyces cerevisiae in modulating preeclamptic stress.  


 Background: Preeclampsia is associated with impaired antioxidant defense that results in materno- fetal complications. In addition to antioxidant deficiency, hemolytic disorder has also been observed in preeclamptic mother. Methods: This study aims in analyzing the fetal complications using cord blood RBC (red blood cell); further the antihemolytic and antioxidant efficiency of two common probiotic yeasts Monascus purpureus and Saccharomyces cerevisiae in preeclamptic and normotensive RBCs were assessed. Results: There was a significant decrease in the antioxidant status (p<0.05) with increased oxidative stress, nitrative stress (p<0.05) and hemolysis (p<0.001) in preeclamptic RBC comparatively. M. purpureus demonstrated a highly significant reactive oxygen radical scavenging activity (p<0.001) whereas S. cerevisiae exhibited a highly significant nitric oxide radical scavenging activity (p<0.001). It was noted that oxidative stress hemolysis was decreased with increased antioxidant level in cord blood RBC from both samples after incubation with both yeasts in a similar manner. The antihemolytic property of M. purpureus and S. cerevisiae suggests that S. cerevisiae functions efficiently with increasing stress. Conclusion: This study demonstrates for the first time that despite their differential scavenging activities, a diet rich in M. purpureus and S. cerevisiae could equally serve as a good natural supplement to alleviate the stress status in the preeclamptic fetus (Tab. 4, Fig. 1, Ref. 39). Keywords: Cord blood RBC, hemolysis, Monascus purpureus, Preeclampsia, Saccharomyces cerevisiae. PMID:24020706

Ekambaram, P; Jayachandran, T; Venkatraman, U; Leonard, S



Improved bread-baking process using Saccharomyces cerevisiae displayed with engineered cyclodextrin glucanotransferase.  


A bread-baking process was developed using a potential novel enzyme, cyclodextrin glucanotransferase[3-18] (CGTase[3-18]), that had previously been engineered to have enhanced hydrolyzing activity with little cyclodextrin (CD) formation activity toward starch. CGTase[3-18] was primarily manipulated to be displayed on the cell surface of Saccharomyces cerevisiae. S. cerevisiae carrying pdeltaCGT integrated into the chromosome exhibited starch-hydrolyzing activity at the same optimal pH and temperature as the free enzyme. Volumes of the bread loaves and rice cakes prepared using S. cerevisiae/pdeltaCGT increased by 20% and 45%, respectively, with no detectable CD. Retrogradation rates of the bread and rice cakes decreased significantly during storage. In comparison to the wild type, S. cerevisiae/pdeltaCGT showed improved viability during four freeze-thaw cycles. The results indicated that CGTase[3-18] displayed on the surface of yeast hydrolyzed starch to glucose and maltose that can be used more efficiently for yeast fermentation. Therefore, display of an antistaling enzyme on the cell surface of yeast has potential for enhancing the baking process. PMID:17488117

Shim, Jae-Hoon; Seo, Nam-Seok; Roh, Sun-Ah; Kim, Jung-Wan; Cha, Hyunju; Park, Kwan-Hwa



Regulation of thiamine synthesis in Saccharomyces cerevisiae for improved pyruvate production.  


Metabolic engineering of Saccharomyces cerevisiae for high-yield production of carboxylic acid requires a cytosolic pyruvate pool as precursor. In this study, a novel strategy to improve pyruvate production and reduce metabolic by-products via regulating thiamine synthesis was explored. Two of the thiamine biosynthesis regulatory genes, THI2 and THI3, were disrupted in the S. cerevisiae parent strain FMME-002. The mutants FMME-002?THI2 and FMME-002?THI3 both exhibited an enhanced pyruvate yield. Moreover, FMME-002?THI2 achieved a relatively higher pyruvate production, and the highest concentration of pyruvate was achieved when 0.04?µ m thiamine was added. Enzyme assays and fermentation profiles of the THI2-complemented strain indicated that the observed metabolic changes represented intrinsic effects of THI2 deletion on the physiology of S. cerevisiae. Under optimal C:N ratio conditions, FMME-002?THI2 produced pyruvate up to 8.21?±?0.30?g/l, whereas the ethanol titre decreased to 2.21?±?0.24?g/l after 96?h of cultivation. These results demonstrate the possibility of improving pyruvate production by regulating thiamine synthesis in S. cerevisiae. PMID:22674684

Xu, Guoqiang; Hua, Qiang; Duan, Ningjun; Liu, Liming; Chen, Jian



Modulation of Promoter and Secretion Efficiency for Improved Heterologous Gene Expression in the Yeast 'Saccharomyces cerevisiae'.  

National Technical Information Service (NTIS)

The yeast Saccharomyces cerevisiae has certain advantages as a host for the production of heterologous proteins. It is a eukaryote, in which the production of proteins of higher eukaryotes may be more successful than in bacterial cells. Specific posttrans...

L. Ruohonen



Effects of the RAD52 Gene on Recombination in Saccharomyces Cerevisiae.  

National Technical Information Service (NTIS)

Effects of the rad52 mutation in Saccharomyces cerevisiae on meiotic, gamma -ray-induced, uv-induced, and spontaneous mitotic recombination were studied. The rad52/rad52 diploids undergo premeiotic DNA synthesis; sporulation occurs but inviable spores are...

S. Prakash L. Prakash W. Burke B. A. Montelone



Presence and Regulation of the Synthesis of Two Alcohol Dehydrogenases from Saccharomyces Cerevisiae.  

National Technical Information Service (NTIS)

Saccharomyces cerevisiae is able to synthesize two different alcohol dehydrogenases according to the culture carbon substrate: the alcohol dehydrogenase 1 produced by the yeast grown on glucose is the fermentative alcohol dehydrogenase, and the alcohol 2 ...

L. Schimpfessel



A metacaspase of Trypanosoma brucei causes loss of respiration competence and clonal death in the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

Metacaspases constitute a new group of cysteine proteases homologous to caspases. Heterologous expression of Trypanosoma brucei metacaspase TbMCA4 in the budding yeast Saccharomyces cerevisiae resulted in growth inhibition, mitochondrial dysfunction and clonal death. The metacaspase orthologue of yeast, ScMCA1 (YOR197w), exhibited genetic interaction with WWM1 (YFL010c), which encodes a small WW domain protein. WWM1 overexpression resulted in growth arrest and

Alexander Szallies; Bruno K. Kubata; Michael Duszenko



Novel interaction of the Hsp90 chaperone machine with Ssl2, an essential DNA helicase in Saccharomyces cerevisiae  

Microsoft Academic Search

Hsp90 is an essential molecular chaperone that is critical for the activity of diverse cellular proteins. Hsp90 functions with a number of co-chaperone proteins, including Sti1\\/Hop. We conducted a genetic screen in Saccharomyces cerevisiae to isolate mutations that exhibit enhanced growth defects in the absence of STI1. We obtained mutations in genes encoding components of the Hsp90 chaperone machine, HSC82,

Gary Flom; Jared Weekes; Jill L. Johnson



The Influence of Microgravity on Invasive Growth in Saccharomyces cerevisiae  

NASA Astrophysics Data System (ADS)

This study investigates the effects of microgravity on colony growth and the morphological transition from single cells to short invasive filaments in the model eukaryotic organism Saccharomyces cerevisiae. Two-dimensional spreading of the yeast colonies grown on semi-solid agar medium was reduced under microgravity in the ?1278b laboratory strain but not in the CMBSESA1 industrial strain. This was supported by the ?1278b proteome map under microgravity conditions, which revealed upregulation of proteins linked to anaerobic conditions. The ?1278b strain showed a reduced invasive growth in the center of the yeast colony. Bud scar distribution was slightly affected, with a switch toward more random budding. Together, microgravity conditions disturb spatially programmed budding patterns and generate strain-dependent growth differences in yeast colonies on semi-solid medium.

Van Mulders, Sebastiaan E.; Stassen, Catherine; Daenen, Luk; Devreese, Bart; Siewers, Verena; van Eijsden, Rudy G. E.; Nielsen, Jens; Delvaux, Freddy R.; Willaert, Ronnie



Mutants of Saccharomyces cerevisiae with defective vacuolar function  

SciTech Connect

Mutants of the yeast Saccharomyces cerevisiae that have a small vacuolar lysine pool were isolated and characterized. Mutant KL97 (lys1 slp1-1) and strain KL197-1A (slp1-1), a prototrophic derivative of KL97, did not grow well in synthetic medium supplemented with 10 mM lysine. Genetic studies indicated that the slp1-1mutation (for small lysine pool) is recessive and is due to a single chromosomal mutation. Mutant KL97 shows the following pleiotropic defects in vacuolar functions. (i) It has small vacuolar pools for lysine, arginine, and histidine. (ii) Its growth is sensitive to lysine, histidine, Ca/sup 2 +/, heavy metal ions, and antibiotics. (iii) It has many small vesicles but no large central vacuole. (iv) It has a normal amount of the vacuolar membrane marker ..cap alpha..-mannosidase but shows reduced activities of the vacuole sap markers proteinase A, proteinase B, and carboxypeptidase Y.

Kitamoto, K.; Yoshizawa, K.; Ohsumi, Y.; Anraku, Y.



Polyadenylation of rRNA in Saccharomyces cerevisiae.  


In contrast to mRNAs, rRNAs are transcribed by RNA polymerase I or III and are not believed to be polyadenylated. Here we show that in Saccharomyces cerevisiae, at least a small fraction of rRNAs do have a poly(A) tail. The levels of polyadenylated rRNAs are dramatically increased in strains lacking the degradation function of Rrp6p, a component of the nuclear exosome. Pap1p, the poly(A) polymerase, is responsible for adenylating the rRNAs despite the fact that the rRNAs do not have a canonical polyadenylation signal. Polyadenylated rRNAs reside mainly within the nucleus and are in turn degraded. For at least one rRNA type, the polyadenylation preferentially occurs on the precursor rather than the mature product. The existence of polyadenylated rRNAs may reflect a quality-control mechanism of rRNA biogenesis. PMID:15173578

Kuai, Letian; Fang, Feng; Butler, J Scott; Sherman, Fred



Preferentially Quantized Linker DNA Lengths in Saccharomyces cerevisiae  

PubMed Central

The exact lengths of linker DNAs connecting adjacent nucleosomes specify the intrinsic three-dimensional structures of eukaryotic chromatin fibers. Some studies suggest that linker DNA lengths preferentially occur at certain quantized values, differing one from another by integral multiples of the DNA helical repeat, ?10 bp; however, studies in the literature are inconsistent. Here, we investigate linker DNA length distributions in the yeast Saccharomyces cerevisiae genome, using two novel methods: a Fourier analysis of genomic dinucleotide periodicities adjacent to experimentally mapped nucleosomes and a duration hidden Markov model applied to experimentally defined dinucleosomes. Both methods reveal that linker DNA lengths in yeast are preferentially periodic at the DNA helical repeat (?10 bp), obeying the forms 10n+5 bp (integer n). This 10 bp periodicity implies an ordered superhelical intrinsic structure for the average chromatin fiber in yeast.

Wang, Ji-Ping; Fondufe-Mittendorf, Yvonne; Xi, Liqun; Tsai, Guei-Feng; Segal, Eran; Widom, Jonathan



Regulation of Phospholipid Synthesis in the Yeast Saccharomyces cerevisiae  

PubMed Central

The yeast Saccharomyces cerevisiae, with its full complement of organelles, synthesizes membrane phospholipids by pathways that are generally common to those found in higher eukaryotes. Phospholipid synthesis in yeast is regulated in response to a variety of growth conditions (e.g., inositol supplementation, zinc depletion, and growth stage) by a coordination of genetic (e.g., transcriptional activation and repression) and biochemical (e.g., activity modulation and localization) mechanisms. Phosphatidate (PA), whose cellular levels are controlled by the activities of key phospholipid synthesis enzymes, plays a central role in the transcriptional regulation of phospholipid synthesis genes. In addition to the regulation of gene expression, phosphorylation of key phospholipid synthesis catalytic and regulatory proteins controls the metabolism of phospholipid precursors and products.

Carman, George M.; Han, Gil-Soo



Architecture and Biosynthesis of the Saccharomyces cerevisiae Cell Wall  

PubMed Central

The wall gives a Saccharomyces cerevisiae cell its osmotic integrity; defines cell shape during budding growth, mating, sporulation, and pseudohypha formation; and presents adhesive glycoproteins to other yeast cells. The wall consists of ?1,3- and ?1,6-glucans, a small amount of chitin, and many different proteins that may bear N- and O-linked glycans and a glycolipid anchor. These components become cross-linked in various ways to form higher-order complexes. Wall composition and degree of cross-linking vary during growth and development and change in response to cell wall stress. This article reviews wall biogenesis in vegetative cells, covering the structure of wall components and how they are cross-linked; the biosynthesis of N- and O-linked glycans, glycosylphosphatidylinositol membrane anchors, ?1,3- and ?1,6-linked glucans, and chitin; the reactions that cross-link wall components; and the possible functions of enzymatic and nonenzymatic cell wall proteins.

Orlean, Peter



Cloning of Saccharomyces cerevisiae gene conferring cadmium resistance  

SciTech Connect

A gene conferring resistance to the toxic effect of cadmium ions was cloned out of a Saccharomyces cerevisiae gene library based on vector pL3. Cadmium resistance was phenotypically expressed only if the CAD1 gene was present in a multicopy plasmid. Intrachromosomal integration of the plasmid or insertion of the centromeric region into the plasmid lowered the number of CAD1-gene copies and, correspondingly, the resistance to cadmium ions. The cloned, Sau3A-cleaved fragment of the yeast chromosome was 3.5 kg in size. Restriction analysis and subcloning of different portions of this DNA stretch determined the minimal fragment responsible for the Cad/sup r/ phenotype: the CAD1 gene was found to be located on an XhoI-Sau3A fragment 1.6 kb long. In Escherichia coli, instability of a yeast-DNA region adjacent to the XhoI-Sau3A fragment was observed.

Yanushka, A.P.; Sasnauskas, K.V.; Yanulaitis, A.A.



MLH1 mutations differentially affect meiotic functions in Saccharomyces cerevisiae.  

PubMed Central

To test whether missense mutations in the cancer susceptibility gene MLH1 adversely affect meiosis, we examined 14 yeast MLH1 mutations for effects on meiotic DNA transactions and gamete viability in the yeast Saccharomyces cerevisiae. Mutations analogous to those associated with hereditary nonpolyposis colorectal cancer (HNPCC) or those that reduce Mlh1p interactions with ATP or DNA all impair replicative mismatch repair as measured by increased mutation rates. However, their effects on meiotic heteroduplex repair, crossing over, chromosome segregation, and gametogenesis vary from complete loss of meiotic functions to no meiotic defect, and mutants defective in one meiotic process are not necessarily defective in others. DNA binding and ATP binding but not ATP hydrolysis are required for meiotic crossing over. The results reveal clear separation of different Mlh1p functions in mitosis and meiosis, and they suggest that some, but not all, MLH1 mutations may be a source of human infertility.

Hoffmann, Eva R; Shcherbakova, Polina V; Kunkel, Thomas A; Borts, Rhona H



Water treatment process and system for metals removal using Saccharomyces cerevisiae  


A process and a system for removal of metals from ground water or from soil by bioreducing or bioaccumulating the metals using metal tolerant microorganisms Saccharomyces cerevisiae. Saccharomyces cerevisiae is tolerant to the metals, able to bioreduce the metals to the less toxic state and to accumulate them. The process and the system is useful for removal or substantial reduction of levels of chromium, molybdenum, cobalt, zinc, nickel, calcium, strontium, mercury and copper in water.

Krauter, Paula A. W. (Livermore, CA); Krauter, Gordon W. (Livermore, CA)



Quantifying the individual effects of ethanol and temperature on the fitness advantage of Saccharomyces cerevisiae  

Microsoft Academic Search

The presence of Saccharomyces cerevisiae in grape berries and fresh musts is usually very low. However, as fermentation progresses, the population levels of this species considerably increase. In this study, we use the concept of fitness advantage to measure how increasing ethanol concentrations (0–25%) and temperature values (4–46 °C) in wine fermentations affects competition between S. cerevisiae and several non-Saccharomyces yeasts (Hanseniaspora

Z. Salvadó; F. N. Arroyo-López; E. Barrio; A. Querol; J. M. Guillamón



Variations of two pools of glycogen and carbohydrate in Saccharomyces cerevisiae grown with various ethanol concentrations  

Microsoft Academic Search

Glycogen, a major reservoir of energy in Saccharomyces cerevisiae, is found to be present as soluble and membrane-bound insoluble pools. Yeast cells can store excess glycogen when grown in\\u000a media with higher concentration of sugar or when subjected to nutritional stress conditions. Saccharomyces cerevisiae NCIM-3300 was grown in media having ethanol concentrations up to 12% (v\\/v). The effects of externally

M. S. Dake; J. P. Jadhv; N. B. Patil



Mead production: selection and characterization assays of Saccharomyces cerevisiae strains.  


Mead is a traditional drink, which results from the alcoholic fermentation of diluted honey carried out by yeasts. However, when it is produced in a homemade way, mead producers find several problems, namely, the lack of uniformity in the final product, delayed and arrested fermentations, and the production of "off-flavours" by the yeasts. These problems are usually associated with the inability of yeast strains to respond and adapt to unfavourable and stressful growth conditions. The main objectives of this work were to evaluate the capacity of Saccharomyces cerevisiae strains, isolated from honey of the Trás-os-Montes (Northeast Portugal), to produce mead. Five strains from honey, as well as one laboratory strain and one commercial wine strain, were evaluated in terms of their fermentation performance under ethanol, sulphur dioxide and osmotic stress. All the strains showed similar behaviour in these conditions. Two yeasts strains isolated from honey and the commercial wine strain were further tested for mead production, using two different honey (a dark and a light honey), enriched with two supplements (one commercial and one developed by the research team), as fermentation media. The results obtained in this work show that S. cerevisiae strains isolated from honey, are appropriate for mead production. However it is of extreme importance to take into account the characteristics of the honey, and supplements used in the fermentation medium formulation, in order to achieve the best results in mead production. PMID:19481129

Pereira, Ana Paula; Dias, Teresa; Andrade, João; Ramalhosa, Elsa; Estevinho, Letícia M



Metabolic fluxes regulate the success of sporulation in Saccharomyces cerevisiae.  


In this work we investigated to what extent cellular metabolism and energetics regulate sporulation in Saccharomyces cerevisiae and which metabolic pathways are involved in such regulation. Sporulation, meiosis, and associated metabolic fluxes in S. cerevisiae strain CH1211 were studied in several experimental protocols involving changes of carbon source (acetate, lactate, or pyruvate) or cell density in sporulation medium, or changing the phase of batch growth at which cells were harvested before transfer to sporulation medium. In acetate-based sporulation medium, the rate at which cells utilized glyoxylate and gluconeogenic pathways correlated positively with the percentage of asci per cell at 72 h. In contrast, in lactate sporulation medium the frequency of sporulation correlated negatively with both the rate of lactate consumption and the fluxes through gluconeogenesis and the pyruvate-carboxylase catalyzed step. In the presence of lactate, the respiratory capacity did correlate positively with the percentage of asci per cell. The experimental data suggest that acetate limits fluxes to anabolic precursors during sporulation. In contrast, sporulation on lactate appears to be influenced by catabolic processes or, even more precisely, by the respiratory capacity of yeast cells. The results obtained are discussed in terms of the hypothesis that an imbalance between anabolic and catabolic fluxes may be required for an efficient sporulation. PMID:8549658

Aon, J C; Rapisarda, V A; Cortassa, S



Interactions between adenylyl cyclase, CAP and RAS from Saccharomyces cerevisiae.  


The adenylyl cyclase complex from Saccharomyces cerevisiae contains at least two subunits, a catalytic subunit of M(r) 200,000, encoded by CYR1 and a cyclase associated subunit, of apparent M(r) 70,000, encoded by CAP. The complex is a major effector of RAS proteins in S. cerevisiae. The interactions between CAP, adenylyl cyclase and RAS were explored in a strain of yeast that lacked CAP and contained an epitope tagged adenylyl cyclase. Adenylyl cyclase activity in this strain was not immunoprecipitated with anti-CAP antibodies, but was immunoprecipitated with anti-epitope antibodies. Two anti-CAP polyclonal antisera and five anti-CAP monoclonal antibodies were used in these studies. Like CAP-bound adenylyl cyclase, the CAP-free adenylyl cyclase was fully activated by yeast RAS2. Transformation of cap strains with plasmids expressing portions of CAP allowed the adenylyl cyclase binding sites on CAP to be mapped by immunoprecipitation experiments. In other experiments, deletion mutations of adenylyl cyclase were used to map the CAP binding site on adenylyl cyclase. The adenylyl cyclase binding site localized to the amino one third of CAP (amino acids 1-168), and the CAP binding site localized to the carboxyl terminus of adenylyl cyclase (amino acids 1768-2026). PMID:7531994

Mintzer, K A; Field, J



Sweet wine production by two osmotolerant Saccharomyces cerevisiae strains.  


The use of Saccharomyces cerevisiae to produce sweet wine is difficult because yeast is affected by a hyperosmotic stress due to the high sugar concentrations in the fermenting must. One possible alternative could be the coimmobilization of the osmotolerant yeast strains S. cerevisiae X4 and X5 on Penicillium chrysogenum strain H3 (GRAS) for the partial fermentation of raisin musts. This immobilized has been, namely, as yeast biocapsules. Traditional sweet wine (that is, without fermentation of the must) and must partially fermented by free yeast cells were also used for comparison. Partially fermented sweet wines showed higher concentration of the volatile compounds than traditionally produced wines. The wines obtained by immobilized yeast cells reached minor concentrations of major alcohols than wines by free cells. The consumption of specific nitrogen compounds was dependent on yeast strain and the cellular immobilization. A principal component analysis shows that the compounds related to the response to osmotic stress (glycerol, acetaldehyde, acetoin, and butanediol) clearly differentiate the wines obtained with free yeasts but not the wines obtained with immobilized yeasts. PMID:23601087

García-Martínez, Teresa; de Lerma, Nieves López; Moreno, Juan; Peinado, Rafael A; Millán, M Carmen; Mauricio, Juan C



Production of recombinant Agaricus bisporus tyrosinase in Saccharomyces cerevisiae cells.  


It has been demonstrated that Agaricus bisporus tyrosinase is able to oxidize various phenolic compounds, thus being an enzyme of great importance for a number of biotechnological applications. The tyrosinase-coding PPO2 gene was isolated by reverse-transcription polymerase chain reaction (RT-PCR) using total RNA extracted from the mushroom fruit bodies as template. The gene was sequenced and cloned into pYES2 plasmid, and the resulting pY-PPO2 recombinant vector was then used to transform Saccharomyces cerevisiae cells. Native polyacrylamide gel electrophoresis followed by enzymatic activity staining with L-3,4-dihydroxyphenylalanine (L-DOPA) indicated that the recombinant tyrosinase is biologically active. The recombinant enzyme was overexpressed and biochemically characterized, showing that the catalytic constants of the recombinant tyrosinase were higher than those obtained when a commercial tyrosinase was used, for all the tested substrates. The present study describes the recombinant production of A. bisporus tyrosinase in active form. The produced enzyme has similar properties to the one produced in the native A. bisporus host, and its expression in S. cerevisiae provides good potential for protein engineering and functional studies of this important enzyme. PMID:22996308

Lezzi, Chiara; Bleve, Gianluca; Spagnolo, Stefano; Perrotta, Carla; Grieco, Francesco



Genetic Manipulation of Palmitoylethanolamide Production and Inactivation in Saccharomyces cerevisiae  

PubMed Central

Background Lipids can act as signaling molecules, activating intracellular and membrane-associated receptors to regulate physiological functions. To understand how a newly discovered signaling lipid functions, it is necessary to identify and characterize the enzymes involved in their production and inactivation. The signaling lipid N-palmitoylethanolamine (PEA) is known to activate intracellular and membrane-associated receptors and regulate physiological functions, but little is known about the enzymes involved in its production and inactivation. Principal Findings Here we show that Saccharomyces cerevisiae produce and inactivate PEA, suggesting that genetic manipulations of this lower eukaryote may be used to identify the enzymes involved in PEA metabolism. Accordingly, using single gene deletion mutants, we identified yeast genes that control PEA metabolism, including SPO14 (a yeast homologue of the mammalian phospholipase D) that controls PEA production and YJU3 (a yeast homologue of the mammalian monoacylglycerol lipase) that controls PEA inactivation. We also found that PEA metabolism is affected by heterologous expression of two mammalian proteins involved in neurodegenerative diseases, namely huntingtin and ?-synuclein. Significance Together these findings show that forward and reverse genetics in S. cerevisiae can be used to identify proteins involved in PEA production and inactivation, and suggest that mutated proteins causing neurodegenerative diseases might affect the metabolism of this important signaling lipid.

Muccioli, Giulio G.; Sia, Angela; Muchowski, Paul J.; Stella, Nephi



Regulation of partitioned sterol biosynthesis in Saccharomyces cerevisiae.  

PubMed Central

Using yeast strains with null mutations in structural genes which encode delta-aminolevulinic acid synthetase (HEM1), isozymes of 3-hydroxy-3-methylglutaryl coenzyme A (HMG1 and HMG2), squalene epoxidase (ERG1), and fatty acid delta 9-desaturase (OLE1), we were able to determine the effect of hemes, sterols, and unsaturated fatty acids on both sterol production and the specific activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) in Saccharomyces cerevisiae. We found that the HMGR isozymes direct essentially equal amounts of carbon to the biosynthesis of sterols under heme-competent conditions, despite a huge disparity (57-fold) in the specific activities of the reductases. Our results demonstrate that palmitoleic acid (16:1) acts as a rate-limiting positive regulator and that ergosterol acts as a potent inhibitor of sterol production in strains which possess only the HMGR1 isozyme (HMG1 hmg2). In strains which contain only the HMGR2 isozyme (hmg1 HMG2), sterol production was inhibited by oleic acid (18:1) and to a lesser degree by ergosterol. The specific activities of the two reductases (HMGR1 and HMGR2) were found to be differentially regulated by hemes but not by ergosterol, palmitoleic acid, or oleic acid. The disparate effects of unsaturated fatty acids and sterols on these strains lead us to consider the possibility of separate, compartmentalized isoprenoid pathways in S. cerevisiae.

Casey, W M; Keesler, G A; Parks, L W



Exploring improved endoglucanase expression in Saccharomyces cerevisiae strains.  


The endoglucanase I and II genes (egI or Cel7B and egII or Cel5A) of Trichoderma reesei QM6a were successfully cloned and expressed in Saccharomyces cerevisiae under the transcriptional control of the yeast ENO1 promoter and terminator sequences. Random mutagenesis of the egI-bearing plasmid resulted in a twofold increase in extracellular EGI activity. Both endoglucanase genes were co-expressed with the synthetic, codon-optimised cellobiohydrolase gene (s-cbhI) from T. reesei as well as the beta-glucosidase gene (bgl1) from Saccharomycopsis fibuligera in S. cerevisiae. Extracellular endoglucanase activity was lower when co-expressed with s-cbhI or bgl1. Recombinant strains were able to hydrolyse phosphoric acid swollen cellulose, generating mainly cellotriose, cellobiose and glucose. Cellobiose accumulated, suggesting the beta-glucosidase activity to be the rate-limiting factor. As a consequence, the recombinant strains were unable to produce enough glucose for growth on amorphous cellulose. The results of this study provide insight into further optimisation of recombinantly expressed cellulase combinations for saccharification and fermentation of cellulose to ethanol. PMID:20041241

du Plessis, Lisa; Rose, Shaunita H; van Zyl, Willem H



Gene relics in the genome of the yeast Saccharomyces cerevisiae.  


There is increasing evidence that DNA duplication is a common and ongoing process that plays a major role in molecular evolution of genomes and that a large fraction of the duplicated gene copies becomes non-functional by accumulation of deleterious mutations. In order to describe this phenomenon, we systematically searched the 6404 intergenic regions (IRs) of the genome of Saccharomyces cerevisiae for traces of coding sequences presenting degenerated but still recognizable sequence similarity with active open reading frames (5823 annotated ORFs). We detected a total of 124 anciently coding regions, or "gene relics", showing similarity to a total of 149 distinct active ORFs. This set of relics shows a continuum of sequence degeneration from those whose sequence is slightly altered compared to the functional ORF (classically defined as pseudogenes), to those that contains so many deleterious mutations, as to reach the limit of recognition. Gene relics are more concentrated in the subtelomeric regions of the chromosomes, reflecting the high plasticity of these regions. The presence of relics also revealed ancestral duplication events of chromosomal segments that were previously undetected. Some of these segments are intermingled with the more easily recognizable ancestral blocks of duplication, indicating successive duplication events. We present a compilation of all the data available, leading to a total of 278 pseudogenes in the genome of S. cerevisiae. PMID:15194185

Lafontaine, Ingrid; Fischer, Gilles; Talla, Emmanuel; Dujon, Bernard



Identification of a Saccharomyces cerevisiae Glucosidase That Hydrolyzes Flavonoid Glucosides? †  

PubMed Central

Baker's yeast (Saccharomyces cerevisiae) whole-cell bioconversions of naringenin 7-O-?-glucoside revealed considerable ?-glucosidase activity, which impairs any strategy to generate or modify flavonoid glucosides in yeast transformants. Up to 10 putative glycoside hydrolases annotated in the S. cerevisiae genome database were overexpressed with His tags in yeast cells. Examination of these recombinant, partially purified polypeptides for hydrolytic activity with synthetic chromogenic ?- or ?-glucosides identified three efficient ?-glucosidases (EXG1, SPR1, and YIR007W), which were further assayed with natural flavonoid ?-glucoside substrates and product verification by thin-layer chromatography (TLC) or high-performance liquid chromatography (HPLC). Preferential hydrolysis of 7- or 4?-O-glucosides of isoflavones, flavonols, flavones, and flavanones was observed in vitro with all three glucosidases, while anthocyanins were also accepted as substrates. The glucosidase activities of EXG1 and SPR1 were completely abolished by Val168Tyr mutation, which confirmed the relevance of this residue, as reported for other glucosidases. Most importantly, biotransformation experiments with knockout yeast strains revealed that only EXG1 knockout strains lost the capability to hydrolyze flavonoid glucosides.

Schmidt, Sabine; Rainieri, Sandra; Witte, Simone; Matern, Ulrich; Martens, Stefan



Mismatch Correction Acts as a Barrier to Homeologous Recombination in Saccharomyces Cerevisiae  

PubMed Central

A homeologous mitotic recombination assay was used to test the role of Saccharomyces cerevisiae mismatch repair genes PMS1, MSH2 and MSH3 on recombination fidelity. A homeologous gene pair consisting of S. cerevisiae SPT15 and its S. pombe homolog were present as a direct repeat on chromosome V, with the exogenous S. pombe sequences inserted either upstream or downstream of the endogenous S. cerevisiae gene. Each gene carried a different inactivating mutation, rendering the starting strain Spt15(-). Recombinants that regenerated SPT15 function were scored after nonselective growth of the cells. In strains wild type for mismatch repair, homeologous recombination was depressed 150- to 180-fold relative to homologous controls, indicating that recombination between diverged sequences is inhibited. In one orientation of the homeologous gene pair, msh2 or msh3 mutations resulted in 17- and 9.6-fold elevations in recombination and the msh2 msh3 double mutant exhibited an 43-fold increase, implying that each MSH gene can function independently in trans to prevent homeologous recombination. Homologous recombination was not significantly affected by the msh mutations. In the other orientation, only msh2 strains were elevated (12-fold) for homeologous recombination. A mutation in MSH3 did not affect the rate of recombination in this orientation. Surprisingly, a pms1 deletion mutant did not exhibit elevated homeologous recombination.

Selva, E. M.; New, L.; Crouse, G. F.; Lahue, R. S.



Influence of temperature and nutrient strength on the susceptibility of Saccharomyces cerevisiae to heavy metals  

SciTech Connect

Saccharomyces cerevisiae is not only a key microorganism in brewing or fermentation processes, it has also been employed for monitoring aquatic pollutants. The major advantage of using Saccharomyces cerevisiae as a bioassay system is that this yeast can be easily obtained as dry pellets from commercial sources at low cost. In addition to its economical aspect, Saccharomyces cerevisiae, like other microorganisms, is easy to handle, grows rapidly, and provides a large number of homogeneous individuals for utilization in toxicity tests. Although cell growth, cell viability, electron transport and mitochondrial respiration of Saccharomyces cerevisiaes have all been selected as parameters for toxicity assessment, measuring cell growth by absorbance is by farm the most convenient and rapid method when large amounts of water samples are to be tested. Mochida et al. (1988), however, reported that Saccharomyces cerevisiae was five to ten times less sensitive than cell culture systems to cadmium, mercury and nickel, when cell growth of both systems was monitored. This relative insensitivity to heavy metals might handicap the practical use of this yeast strain for bioassays. Since previous studies indicated that the susceptibility of microorganisms to environmental toxicants can be influenced by incubation temperature and nutrient strength, we attempted to examine the effect of incubation temperature and nutrient strength on the susceptibility of Saccharomyces cerevisiae to heavy metals in order to obtain the optimum bioassay sensitivity. In this study, we used cadmium and mercury as model toxicants. 9 refs., 2 figs., 1 tab.

Hsu, T.; Lee, L.W.; Chang, T.H. (Development Center for Biotechnology, Taiwan (China))



Non-Coding RNA Prediction and Verification in Saccharomyces cerevisiae  

PubMed Central

Non-coding RNA (ncRNA) play an important and varied role in cellular function. A significant amount of research has been devoted to computational prediction of these genes from genomic sequence, but the ability to do so has remained elusive due to a lack of apparent genomic features. In this work, thermodynamic stability of ncRNA structural elements, as summarized in a Z-score, is used to predict ncRNA in the yeast Saccharomyces cerevisiae. This analysis was coupled with comparative genomics to search for ncRNA genes on chromosome six of S. cerevisiae and S. bayanus. Sets of positive and negative control genes were evaluated to determine the efficacy of thermodynamic stability for discriminating ncRNA from background sequence. The effect of window sizes and step sizes on the sensitivity of ncRNA identification was also explored. Non-coding RNA gene candidates, common to both S. cerevisiae and S. bayanus, were verified using northern blot analysis, rapid amplification of cDNA ends (RACE), and publicly available cDNA library data. Four ncRNA transcripts are well supported by experimental data (RUF10, RUF11, RUF12, RUF13), while one additional putative ncRNA transcript is well supported but the data are not entirely conclusive. Six candidates appear to be structural elements in 5? or 3? untranslated regions of annotated protein-coding genes. This work shows that thermodynamic stability, coupled with comparative genomics, can be used to predict ncRNA with significant structural elements.

Kavanaugh, Laura A.; Dietrich, Fred S.



High level secretion of cellobiohydrolases by Saccharomyces cerevisiae  

PubMed Central

Background The main technological impediment to widespread utilization of lignocellulose for the production of fuels and chemicals is the lack of low-cost technologies to overcome its recalcitrance. Organisms that hydrolyze lignocellulose and produce a valuable product such as ethanol at a high rate and titer could significantly reduce the costs of biomass conversion technologies, and will allow separate conversion steps to be combined in a consolidated bioprocess (CBP). Development of Saccharomyces cerevisiae for CBP requires the high level secretion of cellulases, particularly cellobiohydrolases. Results We expressed various cellobiohydrolases to identify enzymes that were efficiently secreted by S. cerevisiae. For enhanced cellulose hydrolysis, we engineered bimodular derivatives of a well secreted enzyme that naturally lacks the carbohydrate-binding module, and constructed strains expressing combinations of cbh1 and cbh2 genes. Though there was significant variability in the enzyme levels produced, up to approximately 0.3 g/L CBH1 and approximately 1 g/L CBH2 could be produced in high cell density fermentations. Furthermore, we could show activation of the unfolded protein response as a result of cellobiohydrolase production. Finally, we report fermentation of microcrystalline cellulose (Avicel™) to ethanol by CBH-producing S. cerevisiae strains with the addition of beta-glucosidase. Conclusions Gene or protein specific features and compatibility with the host are important for efficient cellobiohydrolase secretion in yeast. The present work demonstrated that production of both CBH1 and CBH2 could be improved to levels where the barrier to CBH sufficiency in the hydrolysis of cellulose was overcome.



Heat shock response improves heterologous protein secretion in Saccharomyces cerevisiae.  


The yeast Saccharomyces cerevisiae is a widely used platform for the production of heterologous proteins of medical or industrial interest. However, heterologous protein productivity is often low due to limitations of the host strain. Heat shock response (HSR) is an inducible, global, cellular stress response, which facilitates the cell recovery from many forms of stress, e.g., heat stress. In S. cerevisiae, HSR is regulated mainly by the transcription factor heat shock factor (Hsf1p) and many of its targets are genes coding for molecular chaperones that promote protein folding and prevent the accumulation of mis-folded or aggregated proteins. In this work, we over-expressed a mutant HSF1 gene HSF1-R206S which can constitutively activate HSR, so the heat shock response was induced at different levels, and we studied the impact of HSR on heterologous protein secretion. We found that moderate and high level over-expression of HSF1-R206S increased heterologous ?-amylase yield 25 and 70 % when glucose was fully consumed, and 37 and 62 % at the end of the ethanol phase, respectively. Moderate and high level over-expression also improved endogenous invertase yield 118 and 94 %, respectively. However, human insulin precursor was only improved slightly and this only by high level over-expression of HSF1-R206S, supporting our previous findings that the production of this protein in S. cerevisiae is not limited by secretion. Our results provide an effective strategy to improve protein secretion and demonstrated an approach that can induce ER and cytosolic chaperones simultaneously. PMID:23208612

Hou, Jin; Osterlund, Tobias; Liu, Zihe; Petranovic, Dina; Nielsen, Jens



Phenotypic Landscape of Saccharomyces cerevisiae during Wine Fermentation: Evidence for Origin-Dependent Metabolic Traits  

Microsoft Academic Search

The species Saccharomyces cerevisiae includes natural strains, clinical isolates, and a large number of strains used in human activities. The aim of this work was to investigate how the adaptation to a broad range of ecological niches may have selectively shaped the yeast metabolic network to generate specific phenotypes. Using 72 S. cerevisiae strains collected from various sources, we provide,

Carole Camarasa; Isabelle Sanchez; Pascale Brial; Frédéric Bigey; Sylvie Dequin; Steven Harris



Epidemiological Investigation of Vaginal Saccharomyces cerevisiae Isolates by a Genotypic Method  

Microsoft Academic Search

Saccharomyces cerevisiae is a ubiquitous, ascomycetous yeast, and vaginitis caused by this organism has been reported only very rarely. The aim of the present investigation was to assess the epidemiological relatedness of a group of vaginal and commercial S. cerevisiae isolates by a previously reported genetic typing method, which divided the isolates into two broad groups with numerous subtypes. Nineteen




Choice of an adequate promoter for efficient complementation in Saccharomyces cerevisiae: a case study  

Microsoft Academic Search

Conservation of the function of open reading frames recently identified in fungal genome projects can be assessed by complementation of deletion mutants of putative Saccharomyces cerevisiae orthologs. A parallel complementation assay expressing the homologous wild type S. cerevisiae gene is generally performed as a positive control. However, we and others have found that failure of complementation can occur in this case.

Libera Lo Presti; Lorenzo Cerutti; Michel Monod; Philippe M. Hauser



Effects of cyclohexane, an industrial solvent, on the yeast Saccharomyces cerevisiae and on isolated yeast mitochondria  

Microsoft Academic Search

Little information on the effects of cyclohexane at the cellular or subcellular level is available. In Saccharomyces cerevisiae, cyclohexane inhibited respiration and diverse energy-dependent processes. In mitochondria isolated from S. cerevisiae, oxygen uptake and ATP synthesis were inhibited, although ATPase activity was not affected. Cyclohexane effects were similar to those reported for beta-pinene and limonene, suggesting that the cyclohexane ring

S. Uribe; P. Rangel; G. Espinola; G. Aguirre



Contribution of winery-resident Saccharomyces cerevisiae strains to spontaneous grape must fermentation  

Microsoft Academic Search

The origin of the Saccharomyces cerevisiae strains that are responsible for spontaneous grape must fermentation was investigated in a long-established industrial winery by means of two different approaches. First, seven selected components of the analytical profiles of the wines produced by 58 strains of S. cerevisiae isolated from different sites and phases of the production cycle of a Grechetto wine

Maurizio Ciani; Ilaria Mannazzu; Paola Marinangeli; Francesca Clementi; Alessandro Martini



The influence of low-intensity ultrasonic on some physiological characteristics of Saccharomyces cerevisiae  

Microsoft Academic Search

In this paper, we studied about some vital physiological characteristics of Saccharomyces cerevisiae at logarithmic phase under ultrasonic stimulation including the stimulated conditions of the 24 kHz in frequency, 2 W in the power efficiency, 1 s for every 15 s in the stimulation time and 30 min in the duration cycle. After S. cerevisiae was inoculated and stimulated for

Shi Lanchun; Wang Bochu; Zhu Liancai; Liu Jie; Yang Yanhong; Duan Chuanren




EPA Science Inventory

We have transformed a Saccharomyces cerevisiae host with an S. cerevisiae genomic library contained in the shuttle vector YEp24 and screened the resultant transformants for resistance to ketoconazole (Kc), an inhibitor of the cytochrome P-450 (P-450) enzyme lanosterol 14-demethyl...


The use of genetically modified Saccharomyces cerevisiae strains in the wine industry  

Microsoft Academic Search

In recent decades, science and food technology have contributed at an accelerated rate to the introduction of new products to satisfy nutritional, socio-economic and quality requirements. With the emergence of modern molecular genetics, the industrial importance of Saccharomyces cerevisiae, is continuously extended. The demand for suitable genetically modified (GM) S. cerevisiae strains for the biofuel, bakery and beverage industries or

Dorit Schuller; Margarida Casal



Isolation and characterization of Saccharomyces cerevisiae mutants resistant to T-2 toxin  

Microsoft Academic Search

T-2 toxin, a trichothecene mycotoxin, inhibits the growth of Saccharomyces cerevisiae. We have isolated nine spontaneous S. cerevisiae mutants resistant to this toxin. The mutants were distinguished from the wild type according to their degree of resistance to T-2 toxin on media with dextrose or glycerol as the carbon source. Generation time, mutation stability and level of cross-resistance to roridin

H. A. Koshinsky; K. T. Schappert; G. G. Khachatourians



Effects of T-2 toxin on induction of petite mutants and mitochondrial function in Saccharomyces cerevisiae  

Microsoft Academic Search

The influence of the trichothecene mycotoxin T-2 on the mitochondria of Saccharomyces cerevisiae was studied. T-2 is a cytotoxic molecule inhibiting growth and macromolecular synthesis in S. cerevisiae. At low concentrations, T-2 toxin arrested yeast growth on glycerol medium and at higher concentrations, it arrested growth on glucose medium. The toxin was not capable itself of inducing petite mutations. Its

Keith T. Schappert; George G. Khachatourians



Effects of salts on Debaryomyces hansenii and Saccharomyces cerevisiae under stress conditions  

Microsoft Academic Search

The effect of Na+ and K+ on growth and thermal death of Debaryomyces hansenii and Saccharomyces cerevisiae were compared under stress conditions as those commonly found in food environments. At the supraoptimal temperature of 34°C both cations at concentrations of 0.5 M stimulated growth of D. hansenii, while K+ had no effect and Na+ inhibited growth of S. cerevisiae. At

Anabel Almagro; Catarina Prista; Santiago Castro; Célia Quintas; Amândio Madeira-Lopes; José Ramos; Maria C Loureiro-Dias



Hxt-Carrier-Mediated Glucose Efflux upon Exposure of Saccharomyces cerevisiae to Excess Maltose  

Microsoft Academic Search

When wild-type Saccharomyces cerevisiae strains pregrown in maltose-limited chemostat cultures were ex- posed to excess maltose, release of glucose into the external medium was observed. Control experiments confirmed that glucose release was not caused by cell lysis or extracellular maltose hydrolysis. To test the hypothesis that glucose efflux involved plasma membrane glucose transporters, experiments were performed with an S. cerevisiae

Mickel L. A. Jansen; Johannes H. De Winde; Jack T. Pronk



Physiological aspects of growth and recombinant DNA stability in Saccharomyces cerevisiae  

Microsoft Academic Search

Despite the fact that plasmid stability in the yeastSaccharomyces cerevisiae is influenced by both genetical and physiological parameters most attention has been focussed on the former. Physiological factors affecting the stability of plasmids have been poorly characterized despite the need for such information in order to optimize the use ofS. cerevisiae as a host for recombinant protein production processes. The

C. Anthony Mason



Saccharomyces cerevisiae: Population Divergence and Resistance to Oxidative Stress in Clinical, Domesticated and Wild Isolates  

Microsoft Academic Search

BackgroundSaccharomyces cerevisiae has been associated with human life for millennia in the brewery and bakery. Recently it has been recognized as an emerging opportunistic pathogen. To study the evolutionary history of S. cerevisiae, the origin of clinical isolates and the importance of a virulence-associated trait, population genetics and phenotypic assays have been applied to an ecologically diverse set of 103

Stephanie Diezmann; Fred S. Dietrich; Justin C. Fay



Genome-wide expression analyses: Metabolic adaptation of Saccharomyces cerevisiae to high sugar stress  

Microsoft Academic Search

The transcriptional response of laboratory strains of Saccharomyces cerevisiae to salt or sorbitol stress has been well studied. These studies have yielded valuable data on how the yeast adapts to these stress conditions. However, S. cerevisiae is a saccharophilic fungus and in its natural environment this yeast encounters high concentrations of sugars. For the production of dessert wines, the sugar

Daniel J Erasmus; George K van der Merwe; Hennie J. J van Vuuren



Molecular Characterization of New Natural Hybrids of Saccharomyces cerevisiae and S. kudriavzevii in Brewing  

Microsoft Academic Search

We analyzed 24 beer strains from different origins by using PCR-restriction fragment length polymorphism analysis of different gene regions, and six new Saccharomyces cerevisiae Saccharomyces kudriavzevii hybrid strains were found. This is the first time that the presence in brewing of this new type of hybrid has been demonstrated. From the comparative molecular analysis of these natural hybrids with respect

Sara S. Gonzalez; Eladio Barrio; Amparo Querol



Effects of fermentation temperature on the strain population of Saccharomyces cerevisiae  

Microsoft Academic Search

The influence of fermentation temperature (from 15 to 35 °C) on a mixed strain population was studied. Mitochondrial DNA analysis was used to differentiate Saccharomyces cerevisiae strains and the frequency of each strain during the alcoholic fermentation was determined. The chemical analyses of resulting wines were carried out. The temperature determined how Saccharomyces strains developed and how effectively they fermented.

Ma. Jesús Torija; Nicolas Rozès; Montse Poblet; José Manuel Guillamón; Albert Mas



Effects of fermentation temperature on the strain population of Saccharomyces cerevisiae  

Microsoft Academic Search

The influence of fermentation temperature (from 15 to 35 jC) on a mixed strain population was studied. Mitochondrial DNA analysis was used to differentiate Saccharomyces cerevisiae strains and the frequency of each strain during the alcoholic fermentation was determined. The chemical analyses of resulting wines were carried out. The temperature determined how Saccharomyces strains developed and how effectively they fermented.

Nicolas Rozes; Montse Poblet; Albert Mas


Cisplatin sensitivity in cAMP-dependent protein kinase mutants of Saccharomyces cerevisiae.  


The emergence of cisplatin resistance poses a significant problem to the treatment of a variety of human malignancies. Therefore, understanding the molecular basis of cisplatin resistance could improve the clinical effectiveness of this anticancer agent. Recently, our laboratory has demonstrated that cAMP-dependent protein kinase (PKA) mutants of the Chinese hamster ovary (CHO) and the mouse adrenocortical carcinoma Y1 cells exhibited increased resistance to cisplatin as well as other DNA-damaging drugs. Further studies showed that either the functional inactivation of PKA or the mutation in the regulatory subunit gene may cause increased recognition of cisplatin-damaged DNA and enhanced DNA repair capacity. In this study, we evaluated the role of PKA in modulating cellular sensitivity to cisplatin in a series of PKA mutants of Saccharomyces cerevisiae. Mutants with decreased kinase activity resulting from a srv2 mutation showed no alterations in cisplatin sensitivity. Complementation of TPK1 in a yeast strain containing mutant tpk1 and also tpk2 and tpk3 deletions did not significantly alter its sensitivity to this DNA-damaging agent. Yeast transformants containing increased kinase activity resulting from overexpression of RAS2Val19 or TPK1 and yeast strains having increased kinase activities due to mutations in the BCY1 gene also did not show alterations in their sensitivity to cisplatin. Therefore, results from these studies unambiguously demonstrate that changes in PKA activity have no effect on cisplatin sensitivity in Saccharomyces cerevisiae. PMID:9858882

Cvijic, M E; Yang, W L; Chin, K V


Relatedness of medically important strains of Saccharomyces cerevisiae as revealed by phylogenetics and metabolomics  

Microsoft Academic Search

Ten medically important Saccharomyces strains, comprising six clinical isolates of Saccharomyces cerevisiae and four probiotic strains of Saccharomyces boulardii ,w ere characterized at the genetic and metabolic level and compared with non-medical, commercial yeast strains used in baking and wine-making. Strains were compared by genetic fingerprinting using amplified fragment length polymorphism (AFLP) analysis, by ribosomal DNA ITS1 sequencing and by

Donald A. MacKenzie; Marianne Defernez; Warwick B. Dunn; Marie Brown; Linda J. Fuller; Andreas Günther; Steve A. James; John Eagles; Mark Philo; Royston Goodacre; Ian N. Roberts



Relationship between sugar uptake kinetics and total sugar consumption in different industrial Saccharomyces cerevisiae strains during alcoholic fermentation  

Microsoft Academic Search

Summary The sugar transport, fermentative alcohol dehydrogenase (ADH) and protease activities of different industrial strains ofSaccharomyces cerevisiae were measured during batch alcoholic fermentation.

J. M. Salmon; J. C. Mauricio



Comparison of pyruvate decarboxylases from Saccharomyces cerevisiae and Komagataella pastoris (Pichia pastoris).  


Pyruvate decarboxylases (PDCs) are a class of enzymes which carry out the non-oxidative decarboxylation of pyruvate to acetaldehyde. These enzymes are also capable of carboligation reactions and can generate chiral intermediates of substantial pharmaceutical interest. Typically, the decarboxylation and carboligation processes are carried out using whole cell systems. However, fermentative organisms such as Saccharomyces cerevisiae are known to contain several PDC isozymes; the precise suitability and role of each of these isozymes in these processes is not well understood. S. cerevisiae has three catalytic isozymes of pyruvate decarboxylase (ScPDCs). Of these, ScPDC1 has been investigated in detail by various groups with the other two catalytic isozymes, ScPDC5 and ScPDC6 being less well characterized. Pyruvate decarboxylase activity can also be detected in the cell lysates of Komagataella pastoris, a Crabtree-negative yeast, and consequently it is of interest to investigate whether this enzyme has different kinetic properties. This is also the first report of the expression and functional characterization of pyruvate decarboxylase from K. pastoris (PpPDC). This investigation helps in understanding the roles of the three isozymes at different phases of S. cerevisiae fermentation as well as their relevance for ethanol and carboligation reactions. The kinetic and physical properties of the four isozymes were determined using similar conditions of expression and characterization. ScPDC5 has comparable decarboxylation efficiency to that of ScPDC1; however, the former has the highest rate of reaction, and thus can be used for industrial production of ethanol. ScPDC6 has the least decarboxylation efficiency of all three isozymes of S. cerevisiae. PpPDC in comparison to all isozymes of S. cerevisiae is less efficient at decarboxylation. All the enzymes exhibit allostery, indicating that they are substrate activated. PMID:23423327

Agarwal, Praveen Kumar; Uppada, Vanita; Noronha, Santosh B



Genome-wide analysis of the effects of location and number of stress response elements on gene expression in Saccharomyces cerevisiae.  


We analyzed the effects of the location and number of stress response elements (STREs) on gene expression in Saccharomyces cerevisiae. Genes containing STRE between 51 and 300 bp upstream from translational start codon tended to be up-regulated and genes with multiple STREs exhibited higher up-regulation under stress conditions. PMID:19111649

Yoshikawa, Katsunori; Furusawa, Chikara; Hirasawa, Takashi; Shimizu, Hiroshi



V-ATPase dysfunction suppresses polyphosphate synthesis in Saccharomyces cerevisiae.  


The yeast Saccharomyces cerevisiae accumulates the high levels of inorganic polyphosphates (polyPs) performing in the cells numerous functions, including phosphate and energy storage. The effects of vacuolar membrane ATPase (V-ATPase) dysfunction were studied on polyP accumulation under short-term cultivation in the Pi-excess media after Pi starvation. The addition of bafilomycin A1, a specific inhibitor of V-ATPase, to the medium with glucose resulted in strong inhibition of the synthesis of long-chain polyP and in substantial suppression of short-chain polyP. The addition of bafilomycin to the medium with ethanol resulted in decreased accumulation of high-molecular polyP, while the accumulation of low-molecular polyP was not affected. The levels of polyP synthesis in the mutant strain with a deletion in the vma2 gene encoding a V-ATPase subunit were significantly lower than in the parent strain in the media with glucose and with ethanol. The synthesis of the longest chain polyP was not observed in the mutant cells. The synthesis of only the low-polymer acid-soluble polyP fraction occurred in the cells of the mutant strain. However, the level of polyP1 was nearly tenfold lower than compared to the cells of the parent strain. Both bafilomycin A1 and the mutation in vacuolar ATPase subunit vma2 lead to a considerable decrease of cellular polyP accumulation. Thus, the defects in ??H(+) formation on the vacuolar membrane resulted in the decrease of polyP biosynthesis in S. cerevisiae. PMID:23371743

Trilisenko, Ludmila; Tomashevsky, Alexander; Kulakovskaya, Tatiana; Kulaev, Igor



Isolation and characterization of a Saccharomyces cerevisiae peptide transport gene.  

PubMed Central

We have cloned and characterized a Saccharomyces cerevisiae peptide transport gene (PTR2) isolated from a genomic DNA library by directly selecting for functional complementation of a peptide transport-deficient mutant. Deletion and frameshift mutageneses were used to localize the complementing activity to a 3.1-kbp region on the transforming plasmid. DNA sequencing of the complementing region identified an open reading frame spanning 1,803 bp. The deduced amino acid sequence predicts a hydrophobic peptide consisting of 601 amino acids, having a molecular mass of 68.1 kDa, composed in part of 12 hydrophobic segments, and sharing significant similarities with a nitrate transport protein encoded by the CHL1 gene of Arabidopsis thaliana. Northern (RNA) hybridization experiments demonstrated a single transcript that was 1.8 kb in length and that was transiently induced by the addition of L-leucine to the growth medium. The PTR2 gene was localized to the right arm of chromosome XI by contour-clamped homogeneous electric field gel chromosome blotting and by hybridization to known chromosome XI lambda phage clones of S. cerevisiae DNA. PTR2 was tightly linked to the UBI2 gene, with the coding sequences being separated by a 466-bp region and oriented so that the genes were transcribed convergently. A chromosomal disruption of the PTR2 gene in a haploid strain was not lethal under standard growth conditions. The cloning of PTR2 represents the first example of the molecular genetic characterization of a eucaryotic peptide transport gene. Images

Perry, J R; Basrai, M A; Steiner, H Y; Naider, F; Becker, J M



Saccharomyces cerevisiae RAD2 gene: isolation, subcloning, and partial characterization.  

PubMed Central

A plasmid (pNF2000) containing a 9.7-kilobase pair DNA insert that complements the UV sensitivity of rad2-1, rad2-2, and rad2-4 mutants of Saccharomyces cerevisiae has been isolated from a yeast genomic library. Genetic analysis of strains derived by transformation of rad2 mutants with an integrating plasmid containing a 9.3-kilobase pair fragment from pNF2000 shows that the fragment integrates exclusively at the chromosomal rad2 gene. We therefore conclude that this plasmid contains the RAD2 gene. The 9.3-kilobase pair fragment was partially digested with Sau3A and cloned into a multicopy yeast vector designed for easy retrieval of Sau3A inserts. The smallest subclone that retains the RAD2 gene is 4.5 kilobase pairs. This fragment was partially digested with Sau3A and cloned into an integrating plasmid. These plasmids were isolated and integrated into a heterozygous rad2/RAD2 strain. Plasmids containing internal fragments of the RAD2 gene were identified because they yielded UV-sensitive transformants due to disruption of the RAD2 gene. Sporulation of diploids transformed with integrating plasmids containing internal fragments of RAD2 gave rise to four viable haploids per tetrad, indicating that unlike the RAD3 gene of S. cerevisiae, the RAD2 gene is not essential for the viability of haploid cells under normal growth conditions. Measurements of the RNA transcript by RNA-DNA hybridization with the internal fragment as the probe indicate a size of approximately 3.2 kilobases. Images

Naumovski, L; Friedberg, E C



Effect of cell cycle position on thermotolerance in Saccharomyces cerevisiae.  


We showed that the heat killing curve for exponentially growing Saccharomyces cerevisiae was biphasic. This suggests two populations of cells with different thermal killing characteristics. When exponentially growing cells separated into cell cycle-specific fractions via centrifugal elutriation were heat shocked, the fractions enriched in small unbudded cells showed greater resistance to heat killing than did other cell cycle fractions. Cells arrested as unbudded cells fell into two groups on the basis of thermotolerance. Sulfur-starved cells and the temperature-sensitive mutants cdc25, cdc33, and cdc35 arrested as unbudded cells were in a thermotolerant state. Alpha-factor-treated cells arrested in a thermosensitive state, as did the temperature-sensitive mutant cdc36 when grown at the restrictive temperature. cdc7, which arrested at the G1-S boundary, arrested in a thermosensitive state. Our results suggest that there is a subpopulation of unbudded cells in exponentially growing cultures that is in G0 and not in G1 and that some but not all methods which cause arrest as unbudded cells lead to arrest in G0 as opposed to G1. It has been shown previously that yeast cells acquire thermotolerance to a subsequent challenge at an otherwise lethal temperature during a preincubation at 36 degrees C. We showed that this acquisition of thermotolerance was corrected temporally with a transient increase in the percentage of unbudded cells during the preincubation at 36 degrees C. The results suggest a relationship between the heat shock phenomenon and the cell cycle in S. cerevisiae and relate thermotolerance to transient as well as to more prolonged residence in the G0 state. PMID:3542970

Plesset, J; Ludwig, J R; Cox, B S; McLaughlin, C S



Effect of cell cycle position on thermotolerance in Saccharomyces cerevisiae.  

PubMed Central

We showed that the heat killing curve for exponentially growing Saccharomyces cerevisiae was biphasic. This suggests two populations of cells with different thermal killing characteristics. When exponentially growing cells separated into cell cycle-specific fractions via centrifugal elutriation were heat shocked, the fractions enriched in small unbudded cells showed greater resistance to heat killing than did other cell cycle fractions. Cells arrested as unbudded cells fell into two groups on the basis of thermotolerance. Sulfur-starved cells and the temperature-sensitive mutants cdc25, cdc33, and cdc35 arrested as unbudded cells were in a thermotolerant state. Alpha-factor-treated cells arrested in a thermosensitive state, as did the temperature-sensitive mutant cdc36 when grown at the restrictive temperature. cdc7, which arrested at the G1-S boundary, arrested in a thermosensitive state. Our results suggest that there is a subpopulation of unbudded cells in exponentially growing cultures that is in G0 and not in G1 and that some but not all methods which cause arrest as unbudded cells lead to arrest in G0 as opposed to G1. It has been shown previously that yeast cells acquire thermotolerance to a subsequent challenge at an otherwise lethal temperature during a preincubation at 36 degrees C. We showed that this acquisition of thermotolerance was corrected temporally with a transient increase in the percentage of unbudded cells during the preincubation at 36 degrees C. The results suggest a relationship between the heat shock phenomenon and the cell cycle in S. cerevisiae and relate thermotolerance to transient as well as to more prolonged residence in the G0 state.

Plesset, J; Ludwig, J R; Cox, B S; McLaughlin, C S



Introducing a new breed of wine yeast: interspecific hybridisation between a commercial Saccharomyces cerevisiae wine yeast and Saccharomyces mikatae.  


Interspecific hybrids are commonplace in agriculture and horticulture; bread wheat and grapefruit are but two examples. The benefits derived from interspecific hybridisation include the potential of generating advantageous transgressive phenotypes. This paper describes the generation of a new breed of wine yeast by interspecific hybridisation between a commercial Saccharomyces cerevisiae wine yeast strain and Saccharomyces mikatae, a species hitherto not associated with industrial fermentation environs. While commercially available wine yeast strains provide consistent and reliable fermentations, wines produced using single inocula are thought to lack the sensory complexity and rounded palate structure obtained from spontaneous fermentations. In contrast, interspecific yeast hybrids have the potential to deliver increased complexity to wine sensory properties and alternative wine styles through the formation of novel, and wider ranging, yeast volatile fermentation metabolite profiles, whilst maintaining the robustness of the wine yeast parent. Screening of newly generated hybrids from a cross between a S. cerevisiae wine yeast and S. mikatae (closely-related but ecologically distant members of the Saccharomyces sensu stricto clade), has identified progeny with robust fermentation properties and winemaking potential. Chemical analysis showed that, relative to the S. cerevisiae wine yeast parent, hybrids produced wines with different concentrations of volatile metabolites that are known to contribute to wine flavour and aroma, including flavour compounds associated with non-Saccharomyces species. The new S. cerevisiae x S. mikatae hybrids have the potential to produce complex wines akin to products of spontaneous fermentation while giving winemakers the safeguard of an inoculated ferment. PMID:23614011

Bellon, Jennifer R; Schmid, Frank; Capone, Dimitra L; Dunn, Barbara L; Chambers, Paul J



Introducing a New Breed of Wine Yeast: Interspecific Hybridisation between a Commercial Saccharomyces cerevisiae Wine Yeast and Saccharomyces mikatae  

PubMed Central

Interspecific hybrids are commonplace in agriculture and horticulture; bread wheat and grapefruit are but two examples. The benefits derived from interspecific hybridisation include the potential of generating advantageous transgressive phenotypes. This paper describes the generation of a new breed of wine yeast by interspecific hybridisation between a commercial Saccharomyces cerevisiae wine yeast strain and Saccharomyces mikatae, a species hitherto not associated with industrial fermentation environs. While commercially available wine yeast strains provide consistent and reliable fermentations, wines produced using single inocula are thought to lack the sensory complexity and rounded palate structure obtained from spontaneous fermentations. In contrast, interspecific yeast hybrids have the potential to deliver increased complexity to wine sensory properties and alternative wine styles through the formation of novel, and wider ranging, yeast volatile fermentation metabolite profiles, whilst maintaining the robustness of the wine yeast parent. Screening of newly generated hybrids from a cross between a S. cerevisiae wine yeast and S. mikatae (closely-related but ecologically distant members of the Saccharomyces sensu stricto clade), has identified progeny with robust fermentation properties and winemaking potential. Chemical analysis showed that, relative to the S. cerevisiae wine yeast parent, hybrids produced wines with different concentrations of volatile metabolites that are known to contribute to wine flavour and aroma, including flavour compounds associated with non-Saccharomyces species. The new S. cerevisiae x S. mikatae hybrids have the potential to produce complex wines akin to products of spontaneous fermentation while giving winemakers the safeguard of an inoculated ferment.

Bellon, Jennifer R.; Schmid, Frank; Capone, Dimitra L.; Dunn, Barbara L.; Chambers, Paul J.



Outlining a future for non- Saccharomyces yeasts: Selection of putative spoilage wine strains to be used in association with Saccharomyces cerevisiae for grape juice fermentation  

Microsoft Academic Search

The use of non-Saccharomyces yeasts that are generally considered as spoilage yeasts, in association with Saccharomyces cerevisiae for grape must fermentation was here evaluated. Analysis of the main oenological characteristics of pure cultures of 55 yeasts belonging to the genera Hanseniaspora, Pichia, Saccharomycodes and Zygosaccharomyces revealed wide biodiversity within each genus. Moreover, many of these non-Saccharomyces strains had interesting oenological

Paola Domizio; Cristina Romani; Livio Lencioni; Francesca Comitini; Mirko Gobbi; Ilaria Mannazzu; Maurizio Ciani



Improved ethanol fermentation by heterologous endoinulinase and inherent invertase from inulin by Saccharomyces cerevisiae.  


It is hypothesized that introduction of an endoinulinase gene into Saccharomyces cerevisiae will improve its inulin utilization and ethanol fermentation through collaboration between the heterologous endoinulinase and the inherent invertase SUC2. The aim of this work was to test the hypothesis by introducing the endoinulinase gene inuA from Aspergillus niger into S. cerevisiae. The results showed that heterologous inuA expressed in S. cerevisiae selectively digested long chains of inulin into short fructooligosaccharides and parts of these fructooligosaccharides could be efficiently utilized by the yeast. This study demonstrated that collaboration between heterologous endoinulinase and inherent invertase improved inulin degradation and ethanol fermentation in S. cerevisiae. PMID:23683966

Yuan, Bo; Wang, Shi-An; Li, Fu-Li



Competition for glucose between the yeasts Saccharomyces cerevisiae and Candida utilis.  

PubMed Central

The competition between the yeasts Saccharomyces cerevisiae CBS 8066 and Candida utilis CBS 621 for glucose was studied in sugar-limited chemostat cultures. Under aerobic conditions, C. utilis always successfully completed against S. cerevisiae. Only under anaerobic conditions did S. cerevisiae become the dominant species. The rationale behind these observations probably is that under aerobic glucose-limited conditions, high-affinity glucose/proton symporters are present in C. utilis, whereas in S. cerevisiae, glucose transport occurs via facilitated diffusion with low-affinity carriers. Our results explain the frequent occurrence of infections by Crabtree-negative yeasts during bakers' yeast production.

Postma, E; Kuiper, A; Tomasouw, W F; Scheffers, W A; van Dijken, J P



Competition for glucose between the yeasts Saccharomyces cerevisiae and Candida utilis.  


The competition between the yeasts Saccharomyces cerevisiae CBS 8066 and Candida utilis CBS 621 for glucose was studied in sugar-limited chemostat cultures. Under aerobic conditions, C. utilis always successfully completed against S. cerevisiae. Only under anaerobic conditions did S. cerevisiae become the dominant species. The rationale behind these observations probably is that under aerobic glucose-limited conditions, high-affinity glucose/proton symporters are present in C. utilis, whereas in S. cerevisiae, glucose transport occurs via facilitated diffusion with low-affinity carriers. Our results explain the frequent occurrence of infections by Crabtree-negative yeasts during bakers' yeast production. PMID:2694963

Postma, E; Kuiper, A; Tomasouw, W F; Scheffers, W A; van Dijken, J P



Temperature-sensitive glucosamine auxotroph of Saccharomyces cerevisiae.  


Temperature-sensitive revertants were isolated from Saccharomyces cerevisiae D-glucosamine auxotrophs previously obtained in this laboratory (W. L. Whelan and C. E. Ballou, J. Bacteriol. 124:1545-1557, 1975). The auxotrophs lack the enzyme 2-amino-2-deoxy-D-glucose-6-phosphate ketol-isomerase (EC, and the revertants appear to be temperature sensitive in the formation of enzyme activity. The enzyme they produce under permissive conditions decays in activity at a rate comparable to that of the wild-type enzyme, and it has similar kinetic properties. The homozygous diploid mutant fails to sporulate at the nonpermissive temperature. Temperature shift experiments were carried out in an effort to determine what effect glucosamine deficiency had on mannoprotein secretion as reflected in the formation of external asparaginase. Although the results were complicated by the slow decay of the residual ketol-isomerase activity, they did show that mannoprotein synthesis or secretion was altered when the internal pool of D-glucosamine was depleted. PMID:6765596

Ballou, L; Grove, J R; Roon, R J; Wiggs, J; Ballou, C E



Functional and Physical Interactions among Saccharomyces cerevisiae ?-Factor Receptors  

PubMed Central

The ?-factor receptor Ste2p is a G protein-coupled receptor (GPCR) expressed on the surface of MATa haploid cells of the yeast Saccharomyces cerevisiae. Binding of ?-factor to Ste2p results in activation of a heterotrimeric G protein and of the pheromone response pathway. Functional interactions between ?-factor receptors, such as dominant-negative effects and recessive behavior of constitutive and hypersensitive mutant receptors, have been reported previously. We show here that dominant-negative effects of mutant receptors persist over a wide range of ratios of the abundances of G protein to receptor and that such effects are not blocked by covalent fusion of G protein ? subunits to normal receptors. In addition, we detected dominant effects of mutant C-terminally truncated receptors, which had not been previously reported to act in a dominant manner. Furthermore, coexpression of C-terminally truncated receptors with constitutively active mutant receptors results in enhancement of constitutive signaling. Together with previous evidence for oligomerization of Ste2p receptors, these results are consistent with the idea that functional interactions between coexpressed receptors arise from physical interactions between them rather than from competition for limiting downstream components, such as G proteins.

Gehret, Austin U.; Connelly, Sara M.



Dual Effects of Plant Steroidal Alkaloids on Saccharomyces cerevisiae  

PubMed Central

Many plant species accumulate sterols and triterpenes as antimicrobial glycosides. These secondary metabolites (saponins) provide built-in chemical protection against pest and pathogen attack and can also influence induced defense responses. In addition, they have a variety of important pharmacological properties, including anticancer activity. The biological mechanisms underpinning the varied and diverse effects of saponins on microbes, plants, and animals are only poorly understood despite the ecological and pharmaceutical importance of this major class of plant secondary metabolites. Here we have exploited budding yeast (Saccharomyces cerevisiae) to investigate the effects of saponins on eukaryotic cells. The tomato steroidal glycoalkaloid ?-tomatine has antifungal activity towards yeast, and this activity is associated with membrane permeabilization. Removal of a single sugar from the tetrasaccharide chain of ?-tomatine results in a substantial reduction in antimicrobial activity. Surprisingly, the complete loss of sugars leads to enhanced antifungal activity. Experiments with ?-tomatine and its aglycone tomatidine indicate that the mode of action of tomatidine towards yeast is distinct from that of ?-tomatine and does not involve membrane permeabilization. Investigation of the effects of tomatidine on yeast by gene expression and sterol analysis indicate that tomatidine inhibits ergosterol biosynthesis. Tomatidine-treated cells accumulate zymosterol rather than ergosterol, which is consistent with inhibition of the sterol C24 methyltransferase Erg6p. However, erg6 and erg3 mutants (but not erg2 mutants) have enhanced resistance to tomatidine, suggesting a complex interaction of erg mutations, sterol content, and tomatidine resistance.

Simons, Veronika; Morrissey, John P.; Latijnhouwers, Maita; Csukai, Michael; Cleaver, Adam; Yarrow, Carol; Osbourn, Anne



Regulation of the phosphatidylethanolamine methylation pathway in Saccharomyces cerevisiae.  


1. Phosphatidyl-N-methylethanolamine methyltransferase mutants of Saccharomyces cerevisiae were isolated. Genetic analysis showed that phosphatidylethanolamine methyltransferase and phosphatidyl-N-methylethanolamine methyltransferase are coded for by separate genes. Phosphatidyl-N-methylethanolamine methyltransferase activity and phosphatidyl-N,N-dimethylethanolamine methyltransferase activity appeared to be catalyzed by the same enzyme. 2. Phosphatidyl-N-methylethanolamine methyltransferase was found to be repressed by myo-inositol and choline. Both myo-inositol and choline at concentrations of 10 micrograms/ml were required for repression. The decreased enzyme level was restored by the removal of myo-inositol or choline or both. 3. Both myo-inositol and choline were required for the maximum repression of phosphatidylethanolamine methyltransferase in wild-type cells. In contrast, choline was not required for the repression of the enzyme in mutant strain 172. This was due to a single nuclear gene mutation in the genome of strain 172. 4. The activity of the phosphatidylethanolamine methylation pathway in cells decreased with time on incubation of cells with myo-inositol and choline, myo-Inositol could not be replaced by other structurally related compounds, such as scyllo-inositol or mannitol. 5. The physiological significance of the repression of the phosphatidylethanolamine methylation pathway is discussed with respect to the mechanism for maintaining the contents of phosphatidylethanolamine and phosphatidylcholine at normal levels. PMID:6759124

Yamashita, S; Oshima, A; Nikawa, J; Hosaka, K



Coordinated Concentration Changes of Transcripts and Metabolites in Saccharomyces cerevisiae  

PubMed Central

Metabolite concentrations can regulate gene expression, which can in turn regulate metabolic activity. The extent to which functionally related transcripts and metabolites show similar patterns of concentration changes, however, remains unestablished. We measure and analyze the metabolomic and transcriptional responses of Saccharomyces cerevisiae to carbon and nitrogen starvation. Our analysis demonstrates that transcripts and metabolites show coordinated response dynamics. Furthermore, metabolites and gene products whose concentration profiles are alike tend to participate in related biological processes. To identify specific, functionally related genes and metabolites, we develop an approach based on Bayesian integration of the joint metabolomic and transcriptomic data. This algorithm finds interactions by evaluating transcript–metabolite correlations in light of the experimental context in which they occur and the class of metabolite involved. It effectively predicts known enzymatic and regulatory relationships, including a gene–metabolite interaction central to the glycolytic–gluconeogenetic switch. This work provides quantitative evidence that functionally related metabolites and transcripts show coherent patterns of behavior on the genome scale and lays the groundwork for building gene–metabolite interaction networks directly from systems-level data.

Bradley, Patrick H.; Brauer, Matthew J.; Rabinowitz, Joshua D.; Troyanskaya, Olga G.



Transcriptional remodeling in response to iron deprivation in Saccharomyces cerevisiae.  


The budding yeast Saccharomyces cerevisiae responds to depletion of iron in the environment by activating Aft1p, the major iron-dependent transcription factor, and by transcribing systems involved in the uptake of iron. Here, we have studied the transcriptional response to iron deprivation and have identified new Aft1p target genes. We find that other metabolic pathways are regulated by iron: biotin uptake and biosynthesis, nitrogen assimilation, and purine biosynthesis. Two enzymes active in these pathways, biotin synthase and glutamate synthase, require an iron-sulfur cluster for activity. Iron deprivation activates transcription of the biotin importer and simultaneously represses transcription of the entire biotin biosynthetic pathway. Multiple genes involved in nitrogen assimilation and amino acid metabolism are induced by iron deprivation, whereas glutamate synthase, a key enzyme in nitrogen assimilation, is repressed. A CGG palindrome within the promoter of glutamate synthase confers iron-regulated expression, suggesting control by a transcription factor of the binuclear zinc cluster family. We provide evidence that yeast subjected to iron deprivation undergo a transcriptional remodeling, resulting in a shift from iron-dependent to parallel, but iron-independent, metabolic pathways. PMID:14668481

Shakoury-Elizeh, Minoo; Tiedeman, John; Rashford, Jared; Ferea, Tracey; Demeter, Janos; Garcia, Emily; Rolfes, Ronda; Brown, Patrick O; Botstein, David; Philpott, Caroline C



Genetically controlled cell lysis in the yeast Saccharomyces cerevisiae.  


The cell wall of the yeast Saccharomyces cerevisiae is a tough, rigid structure, which presents a significant barrier to the release of native or recombinant proteins from this biotechnologically important organism. There is hence a need to develop inexpensive and efficient methods of lysing yeast cells in order to release their intracellular contents. To develop such a method, a tightly regulated promoter, pMET3, has been used to control three genes involved in cell wall biogenesis: PDE2, SRB1/PSA1, and PKC1. Two of these regulation cassettes, pMET3-SRB1/PSA1 and pMET3-PKC1, have been integrated at the chromosomal loci of the respective genes in order to overcome problems of plasmid instability. Although repression of PDE2 did not cause cell lysis, cells depleted of Srb1p/Psa1p gradually lost their viability and integrity, releasing about 10% of total protein into the medium. Repression of PKC1 led to extensive cell lysis, accompanied by the release of 45% of cellular protein into the medium. A double mutant, carrying both pMET3-SRB1/PSA1 and pMET3-PKC1 cassettes in place of SRB1/PSA1 and PKC1, was constructed and found to permit the efficient release of both homologous and heterologous proteins. © 1999 John Wiley & Sons, Inc., PMID:10404241

Zhang, N; Gardner, D C; Oliver, S G; Stateva, L I



Cellular Memory of Acquired Stress Resistance in Saccharomyces cerevisiae  

PubMed Central

Cellular memory of past experiences has been observed in several organisms and across a variety of experiences, including bacteria “remembering” prior nutritional status and amoeba “learning” to anticipate future environmental conditions. Here, we show that Saccharomyces cerevisiae maintains a multifaceted memory of prior stress exposure. We previously demonstrated that yeast cells exposed to a mild dose of salt acquire subsequent tolerance to severe doses of H2O2. We set out to characterize the retention of acquired tolerance and in the process uncovered two distinct aspects of cellular memory. First, we found that H2O2 resistance persisted for four to five generations after cells were removed from the prior salt treatment and was transmitted to daughter cells that never directly experienced the pretreatment. Maintenance of this memory did not require nascent protein synthesis after the initial salt pretreatment, but rather required long-lived cytosolic catalase Ctt1p that was synthesized during salt exposure and then distributed to daughter cells during subsequent cell divisions. In addition to and separable from the memory of H2O2 resistance, these cells also displayed a faster gene-expression response to subsequent stress at >1000 genes, representing transcriptional memory. The faster gene-expression response requires the nuclear pore component Nup42p and serves an important function by facilitating faster reacquisition of H2O2 tolerance after a second cycle of salt exposure. Memory of prior stress exposure likely provides a significant advantage to microbial populations living in ever-changing environments.

Guan, Qiaoning; Haroon, Suraiya; Bravo, Diego Gonzalez; Will, Jessica L.; Gasch, Audrey P.



Automatic bioprocess control. 4. A prototype batch of Saccharomyces cerevisiae.  


The recent investigations in our high performance bioreactors have shown that living cells can be extremely sensitive to physical-chemical environmental conditions and their changes. Consequently, the relationship bioreactor-living cell must thoroughly be investigated in order to discuss both: whether bioreactor characteristics are limiting/dominating during cultivation and to what extent controlled changes of the cellular environment can lead the cells to a desired physiological state. For these investigations, a generally accepted biological test organism would be helpful, of which the requirements and reactions under certain conditions are well known. Saccharomyces cerevisiae is a well known, very robust but nevertheless sensitive organism, eligible for this purpose. In this article a typical batch cultivation on glucose is presented, collected from approx. 300 experiments. Regarding metabolite production and consumption, seven different phases are distinguished on the basis of approx. 20 sensor signals and their metabolic background is discussed. Prerequisite, however, was an exhaustive knowledge upon extracellular conditions, a task which could successfully be fulfilled with the highly automated equipment introduced in the preceding articles of this series. PMID:7763708

Locher, G; Hahnemann, U; Sonnleitner, B; Fiechter, A



Dependency of size of Saccharomyces cerevisiae cells on growth rate.  

PubMed Central

The mean size and percentage of budded cells of a wild-type haploid strain of Saccharomyces cerevisiae grown in batch culture over a wide range of doubling times (tau) have been measured using microscopic measurements and a particle size analyzer. Mean size increased over a 2.5-fold range with increasing growth rate (from tau = 450 min to tau = 75 min). Mean size is principally a function of growth rate and not of a particular carbon source. The duration of the budded phase increased at slow growth rates according to the empirical equation, budded phase = 0.5 tau + 27 (all in minutes). Using a recent model of the cell cycle in which division is thought to be asymmetric, equations have been derived for mean cell age and mean cell volume. The data are consistent with the notion that initiation of the cell cycle occurs at "start" after attainment of a critical cell size, and this size is dependent on growth rate, being, at slow growth rates, 63% of the volume of fast growth rates. Previous reports are reanalyzed in the light of the unequal division model and associated population equations.

Tyson, C B; Lord, P G; Wheals, A E



Identification of genes affecting vacuole membrane fragmentation in Saccharomyces cerevisiae.  


The equilibrium of membrane fusion and fission influences the volume and copy number of organelles. Fusion of yeast vacuoles has been well characterized but their fission and the mechanisms determining vacuole size and abundance remain poorly understood. We therefore attempted to systematically characterize factors necessary for vacuole fission. Here, we present results of an in vivo screening for deficiencies in vacuolar fragmentation activity of an ordered collection deletion mutants, representing 4881 non-essential genes of the yeast Saccharomyces cerevisiae. The screen identified 133 mutants with strong defects in vacuole fragmentation. These comprise numerous known fragmentation factors, such as the Fab1p complex, Tor1p, Sit4p and the V-ATPase, thus validating the approach. The screen identified many novel factors promoting vacuole fragmentation. Among those are 22 open reading frames of unknown function and three conspicuous clusters of proteins with known function. The clusters concern the ESCRT machinery, adaptins, and lipases, which influence the production of diacylglycerol and phosphatidic acid. A common feature of these factors of known function is their capacity to change membrane curvature, suggesting that they might promote vacuole fragmentation via this property. PMID:23383298

Michaillat, Lydie; Mayer, Andreas



Two distinct subfractions in isolated Saccharomyces cerevisiae plasma membranes.  

PubMed Central

The plasma membrane from Saccharomyces cerevisiae X2180-1A and a secretion-blocked mutant, secl (P. Novick and R. Schekman, Proc. Natl. Acad. Sci. U.S.A. 76:1858-1862, 1979) has been purified. Cell walls were digested by treatment with lyticase followed by concanavalin A coating of spheroplasts. alpha-Methylmannoside treatment after lysis, sonication at high salt concentration, and fractionation on a Renografin gradient resulted in two highly purified membrane fractions sedimenting at densities of 1.15 and 1.17 g/cm3. Yields determined by recovery of vanadate-sensitive ATPase activity were 11 to 18%, and those determined by recovery of the spheroplast surface label 125I were 17 to 29%. Iodinated cells have most of their label in sedimentable, nonspheroplast material. However, both membrane populations contain some 125I surface label and show ATPase activity with pH optima only at 5.5. The apparent Vmax of the plasma membrane ATPase equals 360 to 560 nmol of ATP hydrolyzed per min per mg of protein, with a Km for ATP of 0.7 mM. ATPase specific activity is not decreased in mutant plasma membrane. Analysis of 125I-labeled plasma membrane proteins by two-dimensional gel electrophoresis revealed seven major proteins on the plasma membrane surface. Images

Tschopp, J; Schekman, R



Structural and Functional Analysis of Saccharomyces Cerevisiae Mob1  

SciTech Connect

The Mob proteins function as activator subunits for the Dbf2/Dbf20 family of protein kinases. Human and Xenopus Mob1 protein structures corresponding to the most conserved C-terminal core, but lacking the variable N-terminal region, have been reported and provide a framework for understanding the mechanism of Dbf2/Dbf20 regulation. Here, we report the 2.0 {angstrom} X-ray crystal structure of Saccharomyces cerevisiae Mob1 containing both the conserved C-terminal core and the variable N-terminal region. Within the N-terminal region, three novel structural elements are observed; namely, an {alpha}-helix denoted H0, a strand-like element denoted S0 and a short {beta} strand denoted S-1. Helix H0 associates in an intermolecular manner with a second Mob1 molecule to form a Mob1 homodimer. Strand S0 binds to the core domain in an intramolecular manner across a putative Dbf2 binding site mapped by Mob1 temperature-sensitive alleles and NMR binding experiments. In vivo functional analysis demonstrates that Mob1 mutants that target helix H0 or its reciprocal binding site are biologically compromised. The N-terminal region of Mob1 thus contains structural elements that are functionally important.

Mrkobrada,S.; Boucher, L.; Tyers, D.; Sicheri, F.



Analysis of feedback-resistant anthranilate synthases from Saccharomyces cerevisiae.  

PubMed Central

The initial step of tryptophan biosynthesis is catalyzed by the enzyme anthranilate synthase, which in most microorganisms is subject to feedback inhibition by the end product of the pathway. We have characterized the TRP2 gene from a mutant Saccharomyces cerevisiae strain coding for an anthranilate synthase that is unresponsive to tryptophan. Sequence analysis of this TRP2(Fbr) (feedback-resistant) allele revealed numerous differences from a previously published TRP2 sequence. However, TRP2(Fbr) was found to differ in only one single-point mutation from its own parent wild type, a C-to-T transition resulting in a serine 76-to-leucine 76 amino acid substitution. Therefore, serine 76 is a crucial amino acid for proper regulation of the yeast enzyme. We constructed additional feedback-resistant enzyme forms of the yeast anthranilate synthase by site-directed mutagenesis of the conserved LLES sequence in the TRP2 gene. From analysis of these variants, we propose an extended sequence, LLESX10S, as the regulatory element in tryptophan-responsive anthranilate synthases from prokaryotic and eukaryotic organisms.

Graf, R; Mehmann, B; Braus, G H



Vacuolar Cation/H+ Antiporters of Saccharomyces cerevisiae*  

PubMed Central

We previously demonstrated that Saccharomyces cerevisiae vnx1? mutant strains displayed an almost total loss of Na+ and K+/H+ antiporter activity in a vacuole-enriched fraction. However, using different in vitro transport conditions, we were able to reveal additional K+/H+ antiporter activity. By disrupting genes encoding transporters potentially involved in the vnx1 mutant strain, we determined that Vcx1p is responsible for this activity. This result was further confirmed by complementation of the vnx1?vcx1? nhx1? triple mutant with Vcx1p and its inactivated mutant Vcx1p-H303A. Like the Ca2+/H+ antiporter activity catalyzed by Vcx1p, the K+/H+ antiporter activity was strongly inhibited by Cd2+ and to a lesser extend by Zn2+. Unlike as previously observed for NHX1 or VNX1, VCX1 overexpression only marginally improved the growth of yeast strain AXT3 in the presence of high concentrations of K+ and had no effect on hygromycin sensitivity. Subcellular localization showed that Vcx1p and Vnx1p are targeted to the vacuolar membrane, whereas Nhx1p is targeted to prevacuoles. The relative importance of Nhx1p, Vnx1p, and Vcx1p in the vacuolar accumulation of monovalent cations will be discussed.

Cagnac, Olivier; Aranda-Sicilia, Maria Nieves; Leterrier, Marina; Rodriguez-Rosales, Maria-Pilar; Venema, Kees



Vacuolar cation/H+ antiporters of Saccharomyces cerevisiae.  


We previously demonstrated that Saccharomyces cerevisiae vnx1? mutant strains displayed an almost total loss of Na(+) and K(+)/H(+) antiporter activity in a vacuole-enriched fraction. However, using different in vitro transport conditions, we were able to reveal additional K(+)/H(+) antiporter activity. By disrupting genes encoding transporters potentially involved in the vnx1 mutant strain, we determined that Vcx1p is responsible for this activity. This result was further confirmed by complementation of the vnx1?vcx1? nhx1? triple mutant with Vcx1p and its inactivated mutant Vcx1p-H303A. Like the Ca(2+)/H(+) antiporter activity catalyzed by Vcx1p, the K(+)/H(+) antiporter activity was strongly inhibited by Cd(2+) and to a lesser extend by Zn(2+). Unlike as previously observed for NHX1 or VNX1, VCX1 overexpression only marginally improved the growth of yeast strain AXT3 in the presence of high concentrations of K(+) and had no effect on hygromycin sensitivity. Subcellular localization showed that Vcx1p and Vnx1p are targeted to the vacuolar membrane, whereas Nhx1p is targeted to prevacuoles. The relative importance of Nhx1p, Vnx1p, and Vcx1p in the vacuolar accumulation of monovalent cations will be discussed. PMID:20709757

Cagnac, Olivier; Aranda-Sicilia, Maria Nieves; Leterrier, Marina; Rodriguez-Rosales, Maria-Pilar; Venema, Kees



Identification of genes affecting hydrogen sulfide formation in Saccharomyces cerevisiae.  


A screen of the Saccharomyces cerevisiae deletion strain set was performed to identify genes affecting hydrogen sulfide (H(2)S) production. Mutants were screened using two assays: colony color on BiGGY agar, which detects the basal level of sulfite reductase activity, and production of H(2)S in a synthetic juice medium using lead acetate detection of free sulfide in the headspace. A total of 88 mutants produced darker colony colors than the parental strain, and 4 produced colonies significantly lighter in color. There was no correlation between the appearance of a dark colony color on BiGGY agar and H(2)S production in synthetic juice media. Sixteen null mutations were identified as leading to the production of increased levels of H(2)S in synthetic juice using the headspace analysis assay. All 16 mutants also produced H(2)S in actual juices. Five of these genes encode proteins involved in sulfur containing amino acid or precursor biosynthesis and are directly associated with the sulfate assimilation pathway. The remaining genes encode proteins involved in a variety of cellular activities, including cell membrane integrity, cell energy regulation and balance, or other metabolic functions. The levels of hydrogen sulfide production of each of the 16 strains varied in response to nutritional conditions. In most cases, creation of multiple deletions of the 16 mutations in the same strain did not lead to a further increase in H(2)S production, instead often resulting in decreased levels. PMID:18192430

Linderholm, Angela L; Findleton, Carrie L; Kumar, Gagandeep; Hong, Yeun; Bisson, Linda F



Production of resveratrol from tyrosine in metabolically engineered Saccharomyces cerevisiae.  


Resveratrol, a polyphenol compound found in grape skins, has been proposed to account for the beneficial effects of red wine against heart disease. To produce resveratrol in Saccharomyces cerevisiae, four heterologous genes were introduced: the phenylalanine ammonia lyase gene from Rhodosporidium toruloides, the cinnamic acid 4-hydroxylase and 4-coumarate:coenzyme A ligase genes both from Arabidopsis thaliana, and the stilbene synthase gene from Arachis hypogaea. When this recombinant yeast was cultivated by batch fermentation in YP medium containing 2% galactose, it produced 2.6 mg/L p-coumaric acid and 3.3 mg/L resveratrol. In order to increase the pool of malonyl-CoA, a key precursor in resveratrol biosynthesis, the acetyl-CoA carboxylase (ACC1) gene was additionally overexpressed in the yeast by replacing the native promoter of the ACC1 gene with the stronger GAL1 promoter and this resulted in enhanced production of resveratrol (4.3 mg/L). Furthermore, when tyrosine was supplemented in the medium, the concentration of resveratrol increased up to 5.8 mg/L. This result illustrates a possible strategy for developing metabolically engineered yeast strain for the economical production of resveratrol from cheap amino acids. PMID:22883555

Shin, So-Yeon; Jung, Sang-Min; Kim, Myoung-Dong; Han, Nam Soo; Seo, Jin-Ho



Dissection of filamentous growth by transposon mutagenesis in Saccharomyces cerevisiae.  


Diploid Saccharomyces cerevisiae strains starved for nitrogen undergo a developmental transition from growth as single yeast form (YF) cells to a multicellular form consisting of filaments of pseudohyphal (PH) cells. Filamentous growth is regulated by an evolutionarily conserved signaling pathway that includes the small GTP-binding proteins Ras2p and Cdc42p, the protein kinases Ste20p, Ste11p and Ste7p, and the transcription factor Ste12p. Here, we designed a genetic screen for mutant strains defective for filamentous growth (dfg) to identify novel targets of the filamentation signaling pathway, and we thereby identified 16 different genes, CDC39, STE12, TEC1, WHI3, NAB1, DBR1, CDC55, SRV2, TPM1, SPA2, BNI1, DFG5, DFG9, DFG10, BUD8 and DFG16, mutations that block filamentous growth. Phenotypic analysis of dfg mutant strains genetically dissects filamentous growth into the cellular processes of signal transduction, bud site selection, cell morphogenesis and invasive growth. Epistasis tests between dfg mutant alleles and dominant activated alleles of the RAS2 and STE11 genes, RAS2Val19 and STE11-4, respectively, identify putative targets for the filamentation signaling pathway. Several of the genes described here have homologues in filamentous fungi, where they also regulate fungal development. PMID:9055077

Mösch, H U; Fink, G R



Peroxisomal fatty acid uptake mechanism in Saccharomyces cerevisiae.  


Peroxisomes play a major role in human cellular lipid metabolism, including fatty acid ?-oxidation. The most frequent peroxisomal disorder is X-linked adrenoleukodystrophy, which is caused by mutations in ABCD1. The biochemical hallmark of X-linked adrenoleukodystrophy is the accumulation of very long chain fatty acids (VLCFAs) due to impaired peroxisomal ?-oxidation. Although this suggests a role of ABCD1 in VLCFA import into peroxisomes, no direct experimental evidence is available to substantiate this. To unravel the mechanism of peroxisomal VLCFA transport, we use Saccharomyces cerevisiae as a model organism. Here we provide evidence that in this organism very long chain acyl-CoA esters are hydrolyzed by the Pxa1p-Pxa2p complex prior to the actual transport of their fatty acid moiety into the peroxisomes with the CoA presumably being released into the cytoplasm. The Pxa1p-Pxa2p complex functionally interacts with the acyl-CoA synthetases Faa2p and/or Fat1p on the inner surface of the peroxisomal membrane for subsequent re-esterification of the VLCFAs. Importantly, the Pxa1p-Pxa2p complex shares this molecular mechanism with HsABCD1 and HsABCD2. PMID:22493507

van Roermund, Carlo W T; Ijlst, Lodewijk; Majczak, Wiktor; Waterham, Hans R; Folkerts, Hendrik; Wanders, Ronald J A; Hellingwerf, Klaas J



Production of lipid compounds in the yeast Saccharomyces cerevisiae.  


This review describes progress using the yeast Saccharomyces cerevisiae as a model organism for the fast and efficient analysis of genes and enzyme activities involved in the lipid biosynthetic pathways of several donor organisms. Furthermore, we assess the impact of baker's yeast on the production of novel, high-value lipid compounds. Yeast can be genetically modified to produce selected substances in relatively high amounts. A major advantage in choosing yeast as an object for metabolic engineering is the fact that the lipid pathways in this organism have been described in detail and are well characterized. We focus on the de novo production of three major families of lipid products. These are: (1) sterols, providing some previously known and some novel applications as examples of the lipid pathway enhancement that occurs naturally in yeast, (2) the reconstitution of the biosynthetic pathway of steroid hormones and (3) the biosynthesis of polyunsaturated fatty acids, leading to the biosynthesis of different omega-3 and omega-6 fatty acids which do not occur naturally in yeast. We utilize the current knowledge and point out perspectives and problems for future biotechnological applications in the field of lipid compounds. PMID:14586578

Veen, M; Lang, C



d-Xylulose Fermentation to Ethanol by Saccharomyces cerevisiae  

PubMed Central

We used commercial bakers' yeast (Saccharomyces cerevisiae) to study the conversion of d-xylulose to ethanol in the presence of d-xylose. The rate of ethanol production increased with an increase in yeast cell density. The optimal temperature for d-xylulose fermentation was 35°C, and the optimal pH range was 4 to 6. The fermentation of d-xylulose by yeast resulted in the production of ethanol as the major product; small amounts of xylitol and glycerol were also produced. The production of xylitol was influenced by pH as well as temperature. High pH values and low temperatures enhanced xylitol production. The rate of d-xylulose fermentation decreased when the production of ethanol yielded concentrations of 4% or more. The slow conversion rate of d-xylulose to ethanol was increased by increasing the yeast cell density. The overall production of ethanol from d-xylulose by yeast cells under optimal conditions was 90% of the theoretical yield.

Chiang, Lin-Chang; Gong, Cheng-Shung; Chen, Li-Fu; Tsao, George T.



Saccharomyces cerevisiae HSP70 heat shock elements are functionally distinct.  

PubMed Central

The Saccharomyces cerevisiae HSP70 gene SSA1 has multiple heat shock elements (HSEs). To determine the significance of each of these sequences for expression of SSA1, we analyzed expression from a set of promoters containing point mutations in each of the HSEs, individually and in pairwise combinations. Of the three HSE-like sequences, two (HSE2 and HSE3) were active promoter elements; only one, HSE2, was active under basal growth conditions. Either HSE2 or HSE3 alone was able to drive SSA1 transcription at near-normal rates after heat shock. Both HSE2 and HSE3 were capable of driving basal transcription when placed in the context of the CYC1 promoter. Previous analysis had identified an upstream repressing sequence overlapping HSE2 that repressed basal transcription driven by HSE2. Our analysis showed that basal transcription driven by HSE3 was repressed both by the distant upstream repressing sequence and by closer flanking sequences. The ability to drive basal transcription is not inherent in all natural HSEs, since the HSEs from the heat-inducible SSA3 and SSA4 genes showed no basal activity when placed in the CYC1 vector. Gel mobility shift experiments showed that the same population of heat shock transcription factor molecules bound to HSEs capable of driving basal activity and to HSEs having very low or undetectable basal activity. Images

Young, M R; Craig, E A



Factors involved in anaerobic growth of Saccharomyces cerevisiae.  


Life in the absence of molecular oxygen requires several adaptations. Traditionally, the switch from respiratory metabolism to fermentation has attracted much attention in Saccharomyces cerevisiae, as this is the basis for the use of this yeast in the production of alcohol and in baking. It has also been clear that under anaerobic conditions the yeast is not able to synthesize sterols and unsaturated fatty acids and that for anaerobic growth these have to be added to the media. More recently it has been found that many more factors play a role. Several other biosynthetic reactions also require molecular oxygen and the yeast must have alternatives for these. In addition, the composition of the cell wall and cell membrane show major differences when aerobic and anaerobic cells are compared. All these changes are reflected by the observation that the transcription of more than 500 genes changes significantly between aerobically and anaerobically growing cultures. In this review we will give an overview of the factors that play a role in the survival in the absence of molecular oxygen. PMID:17192845

Ishtar Snoek, I S; Yde Steensma, H



TOR and RAS pathways regulate desiccation tolerance in Saccharomyces cerevisiae  

PubMed Central

Tolerance to desiccation in cultures of Saccharomyces cerevisiae is inducible; only one in a million cells from an exponential culture survive desiccation compared with one in five cells in stationary phase. Here we exploit the desiccation sensitivity of exponentially dividing cells to understand the stresses imposed by desiccation and their stress response pathways. We found that induction of desiccation tolerance is cell autonomous and that there is an inverse correlation between desiccation tolerance and growth rate in glucose-, ammonia-, or phosphate-limited continuous cultures. A transient heat shock induces a 5000–fold increase in desiccation tolerance, whereas hyper-ionic, -reductive, -oxidative, or -osmotic stress induced much less. Furthermore, we provide evidence that the Sch9p-regulated branch of the TOR and Ras-cAMP pathway inhibits desiccation tolerance by inhibiting the stress response transcription factors Gis1p, Msn2p, and Msn4p and by activating Sfp1p, a ribosome biogenesis transcription factor. Among 41 mutants defective in ribosome biogenesis, a subset defective in 60S showed a dramatic increase in desiccation tolerance independent of growth rate. We suggest that reduction of a specific intermediate in 60S biogenesis, resulting from conditions such as heat shock and nutrient deprivation, increases desiccation tolerance.

Welch, Aaron Z.; Gibney, Patrick A.; Botstein, David; Koshland, Douglas E.



Bioflavour production from orange peel hydrolysate using immobilized Saccharomyces cerevisiae.  


The rising trend of bioflavour synthesis by microorganisms is hindered by the high manufacturing costs, partially attributed to the cost of the starting material. To overcome this limitation, in the present study, dilute-acid hydrolysate of orange peel was employed as a low-cost, rich in fermentable sugars substrate for the production of flavour-active compounds by Saccharomyces cerevisiae. With this purpose, the use of immobilized cell technology to protect cells against the various inhibitory compounds present in the hydrolysate was evaluated with regard to yeast viability, carbon and nitrogen consumption and cell ability to produce flavour active compounds. For cell immobilization the encapsulation in Ca alginate beads was used. The results were compared with those obtained using free-cell system. Based on the data obtained immobilized cells showed better growth performance and increased ability for de novo synthesis of volatile esters of "fruity" aroma (phenylethyl acetate, ethyl hexanoate, octanoate, decanoate and dodecanoate) than those of free cells. The potential for in situ production of new formulations containing flavour-active compounds derive from yeast cells and also from essential oil of orange peel (limonene, ?-terpineol) was demonstrated by the fact that bioflavour mixture was found to accumulate within the beads. Furthermore, the ability of the immobilized yeast to perform efficiently repeated batch fermentations of orange peel hydrolysate for bioflavour production was successfully maintained after six consecutive cycles of a total period of 240 h. PMID:23995224

Lalou, Sofia; Mantzouridou, Fani; Paraskevopoulou, Adamantini; Bugarski, Branko; Levic, Steva; Nedovic, Victor



Metabolic functions of duplicate genes in Saccharomyces cerevisiae  

PubMed Central

The roles of duplicate genes and their contribution to the phenomenon of enzyme dispensability are a central issue in molecular and genome evolution. A comprehensive classification of the mechanisms that may have led to their preservation, however, is currently lacking. In a systems biology approach, we classify here back-up, regulatory, and gene dosage functions for the 105 duplicate gene families of Saccharomyces cerevisiae metabolism. The key tool was the reconciled genome-scale metabolic model iLL672, which was based on the older iFF708. Computational predictions of all metabolic gene knockouts were validated with the experimentally determined phenotypes of the entire singleton yeast library of 4658 mutants under five environmental conditions. iLL672 correctly identified 96%-98% and 73%-80% of the viable and lethal singleton phenotypes, respectively. Functional roles for each duplicate family were identified by integrating the iLL672-predicted in silico duplicate knockout phenotypes, genome-scale carbon-flux distributions, singleton mutant phenotypes, and network topology analysis. The results provide no evidence for a particular dominant function that maintains duplicate genes in the genome. In particular, the back-up function is not favored by evolutionary selection because duplicates do not occur more frequently in essential reactions than singleton genes. Instead of a prevailing role, multigene-encoded enzymes cover different functions. Thus, at least for metabolism, persistence of the paralog fraction in the genome can be better explained with an array of different, often overlapping functional roles.

Kuepfer, Lars; Sauer, Uwe; Blank, Lars M.



Genetic Analysis of Default Mating Behavior in Saccharomyces Cerevisiae  

PubMed Central

Haploid Saccharomyces cerevisiae cells find each other during conjugation by orienting their growth toward each other along pheromone gradients (chemotropism). However, when their receptors are saturated for pheromone binding, yeast cells must select a mate by executing a default pathway in which they choose a mating partner at random. We previously demonstrated that this default pathway requires the SPA2 gene. In this report we show that the default mating pathway also requires the AXL1, FUS1, FUS2, FUS3, PEA2, RVS161, and BNI1 genes. These genes, including SPA2, are also important for efficient cell fusion during chemotropic mating. Cells containing null mutations in these genes display defects in cell fusion that subtly affect mating efficiency. In addition, we found that the defect in default mating caused by mutations in SPA2 is partially suppressed by multiple copies of two genes, FUS2 and MFA2. These findings uncover a molecular relationship between default mating and cell fusion. Moreover, because axl1 mutants secrete reduced levels of a-factor and are defective at both cell fusion and default mating, these results reveal an important role for a-factor in cell fusion and default mating. We suggest that default mating places a more stringent requirement on some aspects of cell fusion than does chemotropic mating.

Dorer, R.; Boone, C.; Kimbrough, T.; Kim, J.; Hartwell, L. H.



Tor1 regulates protein solubility in Saccharomyces cerevisiae  

PubMed Central

Accumulation of insoluble protein in cells is associated with aging and aging-related diseases; however, the roles of insoluble protein in these processes are uncertain. The nature and impact of changes to protein solubility during normal aging are less well understood. Using quantitative mass spectrometry, we identify 480 proteins that become insoluble during postmitotic aging in Saccharomyces cerevisiae and show that this ensemble of insoluble proteins is similar to those that accumulate in aging nematodes. SDS-insoluble protein is present exclusively in a nonquiescent subpopulation of postmitotic cells, indicating an asymmetrical distribution of this protein. In addition, we show that nitrogen starvation of young cells is sufficient to cause accumulation of a similar group of insoluble proteins. Although many of the insoluble proteins identified are known to be autophagic substrates, induction of macroautophagy is not required for insoluble protein formation. However, genetic or chemical inhibition of the Tor1 kinase is sufficient to promote accumulation of insoluble protein. We conclude that target of rapamycin complex 1 regulates accumulation of insoluble proteins via mechanisms acting upstream of macroautophagy. Our data indicate that the accumulation of proteins in an SDS-insoluble state in postmitotic cells represents a novel autophagic cargo preparation process that is regulated by the Tor1 kinase.

Peters, Theodore W.; Rardin, Matthew J.; Czerwieniec, Gregg; Evani, Uday S.; Reis-Rodrigues, Pedro; Lithgow, Gordon J.; Mooney, Sean D.; Gibson, Bradford W.; Hughes, Robert E.



gTME for improved xylose fermentation of Saccharomyces cerevisiae.  


Global transcription machinery engineering (gTME) is an approach for reprogramming gene transcription to elicit cellular phenotypes important for technological applications. In our study, the application of gTME to Saccharomyces cerevisiae was to improve xylose utilization and tolerance, which is a key trait for many biofuel programs. Mutation of the transcription factor spt15 was introduced by error-prone polymerase chain reaction and then screened on media using xylose as the sole carbon source. The selected out strain spt15-25 showed modest growth rates in the media containing 50, 100, and 150 g/L of xylose or glucose. Under the following fermentation condition: 30 degrees C, rotating speed of 200 r/min, 500-mL Erlenmeyer flask containing 100-mL media, after 109 h, 93.5% of xylose was consumed in 50 g/L xylose medium. Meanwhile, 98.3% glucose can be metabolized in 50-g/L glucose medium. And the carbon source was 50 g/L glucose-xylose (w/w = 1); the utilization ratio of xylose and glucose was 90.8% and 97.3%, respectively. And all the xylitol concentration was below 2.48 g/L. PMID:19067246

Liu, Hongmei; Yan, Ming; Lai, Cangang; Xu, Lin; Ouyang, Pingkai



Unraveling condition specific gene transcriptional regulatory networks in Saccharomyces cerevisiae  

PubMed Central

Background Gene expression and transcription factor (TF) binding data have been used to reveal gene transcriptional regulatory networks. Existing knowledge of gene regulation can be presented using gene connectivity networks. However, these composite connectivity networks do not specify the range of biological conditions of the activity of each link in the network. Results We present a novel method that utilizes the expression and binding patterns of the neighboring nodes of each link in existing experimentally-based, literature-derived gene transcriptional regulatory networks and extend them in silico using TF-gene binding motifs and a compendium of large expression data from Saccharomyces cerevisiae. Using this method, we predict several hundreds of new transcriptional regulatory TF-gene links, along with experimental conditions in which known and predicted links become active. This approach unravels new links in the yeast gene transcriptional regulatory network by utilizing the known transcriptional regulatory interactions, and is particularly useful for breaking down the composite transcriptional regulatory network to condition specific networks. Conclusion Our methods can facilitate future binding experiments, as they can considerably help focus on the TFs that must be surveyed to understand gene regulation. (Supplemental material and the latest version of the MATLAB implementation of the United Signature Algorithm is available online at [1] or [see Additional files 1, 2, 3, 4, 5, 6, 7, 8, 9, 10])

Kim, Hyunsoo; Hu, William; Kluger, Yuval



Transcriptional response of Saccharomyces cerevisiae to desiccation and rehydration.  


A transcriptional analysis of the response of Saccharomyces cerevisiae strain BY4743 to controlled air-drying (desiccation) and subsequent rehydration under minimal glucose conditions was performed. Expression of genes involved in fatty acid oxidation and the glyoxylate cycle was observed to increase during drying and remained in this state during the rehydration phase. When the BY4743 expression profile for the dried sample was compared to that of a commercially prepared dry active yeast, strikingly similar expression changes were observed. The fact that these two samples, dried by different means, possessed very similar transcriptional profiles supports the hypothesis that the response to desiccation is a coordinated event independent of the particular conditions involved in water removal. Similarities between "stationary-phase-essential genes" and those upregulated during desiccation were also noted, suggesting commonalities in different routes to reduced metabolic states. Trends in extracellular and intracellular glucose and trehalose levels suggested that the cells were in a "holding pattern" during the rehydration phase, a concept that was reinforced by cell cycle analyses. Application of a "redescription mining" algorithm suggested that sulfur metabolism is important for cell survival during desiccation and rehydration. PMID:16332871

Singh, Jatinder; Kumar, Deept; Ramakrishnan, Naren; Singhal, Vibha; Jervis, Jody; Garst, James F; Slaughter, Stephen M; DeSantis, Andrea M; Potts, Malcolm; Helm, Richard F



Metabolic Engineering of the Phenylpropanoid Pathway in Saccharomyces cerevisiae  

PubMed Central

Flavonoids are valuable natural products derived from the phenylpropanoid pathway. The objective of this study was to create a host for the biosynthesis of naringenin, the central precursor of many flavonoids. This was accomplished by introducing the phenylpropanoid pathway with the genes for phenylalanine ammonia lyase (PAL) from Rhodosporidium toruloides, 4-coumarate:coenzyme A (CoA) ligase (4CL) from Arabidopsis thaliana, and chalcone synthase (CHS) from Hypericum androsaemum into two Saccharomyces cerevisiae strains, namely, AH22 and a pad1 knockout mutant. Each gene was cloned and inserted into an expression vector under the control of a separate individual GAL10 promoter. Besides its PAL activity, the recombinant PAL enzyme showed tyrosine ammonia lyase activity, which enabled the biosynthesis of naringenin without introducing cinnamate 4-hydroxylase (C4H). 4CL catalyzed the conversion of both trans-cinnamic acid and p-coumaric acid to their corresponding CoA products, which were further converted to pinocembrin chalcone and naringenin chalcone by CHS. These chalcones were cyclized to pinocembrin and naringenin. The yeast AH22 strain coexpressing PAL, 4CL, and CHS produced approximately 7 mg liter?1 of naringenin and 0.8 mg liter?1 of pinocembrin. Several by-products, such as 2?,4?,6?-trihydroxydihydrochalcone and phloretin, were also identified. Precursor feeding studies indicated that metabolic flux to the engineered flavonoid pathway was limited by the flux to the precursor l-tyrosine.

Jiang, Hanxiao; Wood, Karl V.; Morgan, John A.



Characterization of Alcohol-induced Filamentous Growth in Saccharomyces cerevisiae  

PubMed Central

Diploid cells of the budding yeast Saccharomyces cerevisiae starved for nitrogen differentiate into a filamentous growth form. Poor carbon sources such as starches can also stimulate filamentation, whereas haploid cells undergo a similar invasive growth response in rich medium. Previous work has demonstrated a role for various alcohols, by-products of amino acid metabolism, in altering cellular morphology. We found that several alcohols, notably isoamyl alcohol and 1-butanol, stimulate filamentous growth in haploid cells in which this differentiation is normally repressed. Butanol also induces cell elongation and changes in budding pattern, leading to a pseudohyphal morphology, even in liquid medium. The filamentous colony morphology and cell elongation require elements of the pheromone-responsive MAPK cascade and TEC1, whereas components of the nutrient-sensing machinery, such as MEP2, GPA2, and GPR1, do not affect this phenomenon. A screen for 1-butanol–insensitive mutants identified additional proteins that regulate polarized growth (BUD8, BEM1, BEM4, and FIG1), mitochondrial function (MSM1, MRP21, and HMI1), and a transcriptional regulator (CHD1). Furthermore, we have also found that ethanol stimulates hyperfilamentation in diploid cells, again in a MAPK-dependent manner. Together, these results suggest that yeast may sense a combination of nutrient limitation and metabolic by-products to regulate differentiation.

Lorenz, Michael C.; Cutler, N. Shane; Heitman, Joseph



An overview of membrane transport proteins in Saccharomyces cerevisiae.  


All eukaryotic cells contain a wide variety of proteins embedded in the plasma and internal membranes, which ensure transmembrane solute transport. It is now established that a large proportion of these transport proteins can be grouped into families apparently conserved throughout organisms. This article presents the data of an in silicio analysis aimed at establishing a preliminary classification of membrane transport proteins in Saccharomyces cerevisiae. This analysis was conducted at a time when about 65% of all yeast genes were available in public databases. In addition to approximately 60 transport proteins whose function was at least partially known, approximately 100 deduced protein sequences of unknown function display significant sequence similarity to membrane transport proteins characterized in yeast and/or other organisms. While some protein families have been well characterized by classical genetic experimental approaches, others have largely if not totally escaped characterization. The proteins revealed by this in silicio analysis also include a putative K+ channel, proteins similar to aquaporins of plant and animal origin, proteins similar to Na+-solute symporters, a protein very similar to electroneural cation-chloride cotransporters, and a putative Na+-H+ antiporter. A new research area is anticipated: the functional analysis of many transport proteins whose existence was revealed by genome sequencing. PMID:8720066

Andre, B



MAP Kinase Pathways in the Yeast Saccharomyces cerevisiae  

PubMed Central

A cascade of three protein kinases known as a mitogen-activated protein kinase (MAPK) cascade is commonly found as part of the signaling pathways in eukaryotic cells. Almost two decades of genetic and biochemical experimentation plus the recently completed DNA sequence of the Saccharomyces cerevisiae genome have revealed just five functionally distinct MAPK cascades in this yeast. Sexual conjugation, cell growth, and adaptation to stress, for example, all require MAPK-mediated cellular responses. A primary function of these cascades appears to be the regulation of gene expression in response to extracellular signals or as part of specific developmental processes. In addition, the MAPK cascades often appear to regulate the cell cycle and vice versa. Despite the success of the gene hunter era in revealing these pathways, there are still many significant gaps in our knowledge of the molecular mechanisms for activation of these cascades and how the cascades regulate cell function. For example, comparison of different yeast signaling pathways reveals a surprising variety of different types of upstream signaling proteins that function to activate a MAPK cascade, yet how the upstream proteins actually activate the cascade remains unclear. We also know that the yeast MAPK pathways regulate each other and interact with other signaling pathways to produce a coordinated pattern of gene expression, but the molecular mechanisms of this cross talk are poorly understood. This review is therefore an attempt to present the current knowledge of MAPK pathways in yeast and some directions for future research in this area.

Gustin, Michael C.; Albertyn, Jacobus; Alexander, Matthew; Davenport, Kenneth



Host Factors That Affect Ty3 Retrotransposition in Saccharomyces cerevisiae  

PubMed Central

The retrovirus-like element Ty3 of Saccharomyces cerevisiae integrates at the transcription initiation region of RNA polymerase III. To identify host genes that affect transposition, a collection of insertion mutants was screened using a genetic assay in which insertion of Ty3 activates expression of a tRNA suppressor. Fifty-three loci were identified in this screen. Corresponding knockout mutants were tested for the ability to mobilize a galactose-inducible Ty3, marked with the HIS3 gene. Of 42 mutants tested, 22 had phenotypes similar to those displayed in the original assay. The proteins encoded by the defective genes are involved in chromatin dynamics, transcription, RNA processing, protein modification, cell cycle regulation, nuclear import, and unknown functions. These mutants were induced for Ty3 expression and assayed for Gag3p protein, integrase, cDNA, and Ty3 integration upstream of chromosomal tDNAVal(AAC) genes. Most mutants displayed differences from the wild type in one or more intermediates, although these were typically not as severe as the genetic defect. Because a relatively large number of genes affecting retrotransposition can be identified in yeast and because the majority of these genes have mammalian homologs, this approach provides an avenue for the identification of potential antiviral targets.

Aye, Michael; Irwin, Becky; Beliakova-Bethell, Nadejda; Chen, Eric; Garrus, Jennifer; Sandmeyer, Suzanne



Meiotic Diploid Progeny and Meiotic Nondisjunction in SACCHAROMYCES CEREVISIAE  

PubMed Central

Abnormalities in chromosome number that occurred during meiosis were evaluated with a specially-constructed diploid strain of Saccharomyces cerevisiae. The strain is heterozygous for six markers of the right arm of chromosome V and heterozygous for cyh2 (resistance to cycloheximide) on chromosome VII.—Selection of meiotic spores on a medium containing cycloheximide and required nutrilites—except those for the markers of the right arm of chromosome V—allows the growth of aberrant clones belonging only to two classes: a) diploid clones, caused by failure of the second meiotic division, with a frequency of 0.54 x 10-4 per viable spore; and b) diplo V, aneuploids derived from nondisjunctions in meiosis I or meiosis II, with a total spontaneous frequency of 0.95 x 10-4 per viable spore. About two-thirds of the aneuploids originated during meiosis I, the rest during meiosis II. An investigation of these events in control meioses and after treatment with MMS, Benomyl and Amphotericin B suggests that this assay system is suitable for screening environmental mutagens for their effects on meiotic segregation.

Sora, Silvio; Lucchini, Giovanna; Magni, Giovanni E.



Genetic engineering of taxol biosynthetic genes in Saccharomyces cerevisiae.  


Baccatin III, an intermediate of Taxol biosynthesis and a useful precursor for semisynthesis of the anti-cancer drug, is produced in yew (Taxus) species by a sequence of 15 enzymatic steps from primary metabolism. Ten genes encoding enzymes of this extended pathway have been described, thereby permitting a preliminary attempt to reconstruct early steps of taxane diterpenoid (taxoid) metabolism in Saccharomyces cerevisiae as a microbial production host. Eight of these taxoid biosynthetic genes were functionally expressed in yeast from episomal vectors containing one or more gene cassettes incorporating various epitope tags to permit protein surveillance and differentiation of those pathway enzymes of similar size. All eight recombinant proteins were readily detected by immunoblotting using specific monoclonal antibodies and each expressed protein was determined to be functional by in vitro enzyme assay, although activity levels differed considerably between enzyme types. Using three plasmids carrying different promoters and selection markers, genes encoding five sequential pathway steps leading from primary isoprenoid metabolism to the intermediate taxadien-5alpha- acetoxy-10beta-ol were installed in a single yeast host. Metabolite analysis showed that yeast isoprenoid precursors could be utilized in the reconstituted pathway because products accumulated from the first two engineered pathway steps (leading to the committed intermediate taxadiene); however, a pathway restriction was encountered at the first cytochrome P450 hydroxylation step. The means of overcoming this limitation are described in the context of further development of this novel approach for production of Taxol precursors and related taxoids in yeast. PMID:16161138

Dejong, JingHong M; Liu, Yule; Bollon, Arthur P; Long, Robert M; Jennewein, Stefan; Williams, David; Croteau, Rodney B



Saccharomyces cerevisiae MutL? IS A MISMATCH REPAIR ENDONUCLEASE*  

PubMed Central

MutL homologs are crucial for mismatch repair and genetic stability, but their function is not well understood. Human MutL? (MLH1-PMS2 heterodimer) harbors a latent endonuclease that is dependent on integrity of a PMS2 DQHA(X)2E(X)4E motif (Kadyrov et al. (2006) Cell 126, 297-308). This sequence element is conserved in many MutL homologs, including the PMS1 subunit of Saccharomyces cerevisiae MutL?, but is absent in MutL proteins from bacteria like Escherichia coli that rely on d(GATC) methylation for strand directionality. We show that yeast MutL? is a strand-directed endonuclease that incises DNA in a reaction that depends on a mismatch, yMutS?, yRFC, yPCNA, ATP, and a pre-existing strand break, whereas E. coli MutL is not. Amino acid substitution within the PMS1 DQHA(X)2E(X)4E motif abolishes yMutL? endonuclease activity in vitro and confers strong genetic instability in vivo, but does not affect yMutL? ATPase activity or the ability of the protein to support assembly of the yMutL?•yMutS?•heteroduplex ternary complex. The loaded form of yPCNA may play an important effector role in directing yMutL? incision to the discontinuous strand of a nicked heteroduplex.

Kadyrov, Farid A.; Holmes, Shannon F.; Arana, Mercedes E.; Lukianova, Olga A.; O'Donnell, Mike; Kunkel, Thomas A.; Modrich, Paul



Directed evolution of a highly efficient cellobiose utilizing pathway in an industrial Saccharomyces cerevisiae strain.  


Balancing and increasing the flux through an engineered heterologous pathway in a target organism to achieve high yield and productivity remains an overwhelming challenge in metabolic engineering. Here we report a novel strategy combining directed evolution and promoter engineering for rapid and efficient multi-gene pathway optimization. As proof of concept, this strategy was applied to optimize a cellobiose utilizing pathway in an industrial Saccharomyces cerevisiae strain for highly efficient cellulosic biofuels production. The resulting strain exhibited significantly higher cellobiose consumption rate (6.41-fold) and ethanol productivity (6.36-fold) compared to its parent strain. This study also showed that both the ratios and absolute values of the expression levels of the genes in the cellobiose utilizing pathway play an important role in cellobiose uptake, and ?-glucose is likely one of the key factors affecting cellobiose metabolism. Biotechnol. Biotechnol. Bioeng. 2013;110: 2874-2881. © 2013 Wiley Periodicals, Inc. PMID:23616289

Yuan, Yongbo; Zhao, Huimin



Fluorescent measurement of the intracellular pH during sporulation of Saccharomyces cerevisiae.  


This work reports the intracellular pH (pHi) dynamics of Saccharomyces cerevisiae cells in sporulation medium. Cells loaded with the pH-sensitive dye carboxy-seminaphthorhodafluor-1 (C.SNARF-1) exhibited an alkalization of the pHi following the extracellular pH during sporulation in the absence of buffer and almost no change in pHi or delta pH when sporulation was carried out in buffered medium. The results indicate that the pH gradient does not appear to be directly involved in the regulation of acetate uptake during sporulation. However, the alkalization of pHi by eliciting a decrease in metabolic fluxes could account for a lower demand for acetate. PMID:9252568

Aon, J C; Cortassa, S



Corresponding changes in kynurenine hydroxylase activity, membrane fluidity, and sterol composition in Saccharomyces cerevisiae mitochondria.  

PubMed Central

The effect of sterol composition on the properties of the mitochondrial membrane of Saccharomyces cerevisiae was investigated. The physical state of mitochondrial membranes from wild-type strains and sterol mutants was compared, using a fluorescence polarization technique with 1,6-diphenyl-1,3-5-hexatriene. Changes in the rate of depolarization of the probe molecule as a function of temperature suggest the occurrence of a phase transition in the mitochondrial membranes isolated from the sterol mutants but not in the membranes isolated from the wild types. Arrhenius kinetics of the mitochondrial membrane-bound enzyme L-kynurenine-3-hydroxylase exhibited changes in activation energy at temperatures similar to those observed in the fluorescence polarization study. The ratio of mitochondrial sterol to phospholipid and the phospholipid fatty acid composition of the organisms were characterized.

McLean-Bowen, C A; Parks, L W



Behavior of a fenhexamid photoproduct during the alcoholic fermentation of Saccharomyces cerevisiae.  


The fungicide fenhexamid [N-(2,3-dichloro-4-hydroxyphenyl)-1-methylcyclohexanecarboxamide] degraded rapidly by UV or sunlight irradiation, yielding 7-chloro-6-hydroxy-2-(1-methylcyclohexyl)-1,3-benzoxazole (CHB) as a main photoproduct. CHB was isolated, and its effect on alcoholic fermentation of Saccharomyces cerevisiae was studied. The results indicate that the presence of CHB does not affect the extent of alcohol production. After 12 days, the amount of CHB in the fermentation medium decreased by ca. 65%. Only 25% of the missing CHB was recovered unchanged from yeasts, most likely because it was adsorbed on the yeast wall cell. The remaining part degraded during the fermentation process. Glucan and chitin, two potential adsorbents, which constitute yeast cell walls, exhibited affinity for CHB. PMID:15612795

Cabras, Paolo; Farris, Giovanni A; Pinna, Maria V; Pusino, Alba



Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae  

PubMed Central

Background Hydrocarbon alkanes, components of major fossil fuels, are considered as next-generation biofuels because their biological production has recently been shown to be possible. However, high-yield alkane production requires robust host cells that are tolerant against alkanes, which exhibit cytotoxicity. In this study, we aimed to improve alkane tolerance in Saccharomyces cerevisiae, a key industrial microbial host, by harnessing heterologous transporters that potentially pump out alkanes. Results To this end, we attempted to exploit ABC transporters in Yarrowia lipolytica based on the observation that it utilizes alkanes as a carbon source. We confirmed the increased transcription of ABC2 and ABC3 transporters upon exposure to a range of alkanes in Y. lipolytica. We then showed that the heterologous expression of ABC2 and ABC3 transporters significantly increased tolerance against decane and undecane in S. cerevisiae through maintaining lower intracellular alkane level. In particular, ABC2 transporter increased the tolerance limit of S. cerevisiae about 80-fold against decane. Furthermore, through site-directed mutagenesis for glutamate (E988 for ABC2, and E989 for ABC3) and histidine (H1020 for ABC2, and H1021 for ABC3), we provided the evidence that glutamate was essential for the activity of ABC2 and ABC3 transporters, with ATP most likely to be hydrolyzed by a catalytic carboxylate mechanism. Conclusions Here, we demonstrated that transporter engineering through expression of heterologous efflux pumps led to significantly improved tolerance against alkane biofuels in S. cerevisiae. We believe that our results laid the groundwork for developing robust alkane-producing yeast cells through transporter engineering, which will greatly aid in next-generation alkane biofuel production and recovery.



L-Carnosine Affects the Growth of Saccharomyces cerevisiae in a Metabolism-Dependent Manner  

PubMed Central

The dipeptide L-carnosine (?-alanyl-L-histidine) has been described as enigmatic: it inhibits growth of cancer cells but delays senescence in cultured human fibroblasts and extends the lifespan of male fruit flies. In an attempt to understand these observations, the effects of L-carnosine on the model eukaryote, Saccharomyces cerevisiae, were examined on account of its unique metabolic properties; S. cerevisiae can respire aerobically, but like some tumor cells, it can also exhibit a metabolism in which aerobic respiration is down regulated. L-Carnosine exhibited both inhibitory and stimulatory effects on yeast cells, dependent upon the carbon source in the growth medium. When yeast cells were not reliant on oxidative phosphorylation for energy generation (e.g. when grown on a fermentable carbon source such as 2% glucose), 10–30 mM L-carnosine slowed growth rates in a dose-dependent manner and increased cell death by up to 17%. In contrast, in media containing a non-fermentable carbon source in which yeast are dependent on aerobic respiration (e.g. 2% glycerol), L-carnosine did not provoke cell death. This latter observation was confirmed in the respiratory yeast, Pichia pastoris. Moreover, when deletion strains in the yeast nutrient-sensing pathway were treated with L-carnosine, the cells showed resistance to its inhibitory effects. These findings suggest that L-carnosine affects cells in a metabolism-dependent manner and provide a rationale for its effects on different cell types.

Cartwright, Stephanie P.; Bill, Roslyn M.; Hipkiss, Alan R.



New insights into the Saccharomyces cerevisiae fermentation switch: Dynamic transcriptional response to anaerobicity and glucose-excess  

Microsoft Academic Search

BACKGROUND: The capacity of respiring cultures of Saccharomyces cerevisiae to immediately switch to fast alcoholic fermentation upon a transfer to anaerobic sugar-excess conditions is a key characteristic of Saccharomyces cerevisiae in many of its industrial applications. This transition was studied by exposing aerobic glucose-limited chemostat cultures grown at a low specific growth rate to two simultaneous perturbations: oxygen depletion and

Joost van den Brink; Pascale Daran-Lapujade; Jack T Pronk; Johannes H de Winde



Biophysical characterization of elongin C from Saccharomyces cerevisiae.  


Elongin C (ELC) is an essential component of the mammalian CBC(VHL) E3 ubiquitin ligase complex. As a step toward understanding the role of ELC in assembly and function of CBC-type ubiquitin ligases, we analyzed the quaternary structure and backbone dynamics of the highly homologous Elc1 protein from Saccharomyces cerevisiae. Analytical ultracentrifugation experiments in conjunction with size exclusion chromatography showed that Elc1 is a nonglobular monomer over a wide range of concentrations. Pronounced line broadening in (1)H,(15)N-HSQC NMR spectra and failure to assign peaks corresponding to the carboxy-terminal helix 4 of Elc1 indicated that helix 4 is conformationally labile. Measurement of (15)N NMR relaxation parameters including T(1), T(2), and the (1)H-(15)N nuclear Overhauser effect revealed (i) surprisingly high flexibility of residues 69-77 in loop 5, and (ii) chemical exchange contributions for a large number of residues throughout the protein. Addition of 2,2,2-trifluoroethanol (TFE) stabilized helix 4 and reduced chemical exchange contributions, suggesting that stabilization of helix 4 suppresses the tendency of Elc1 to undergo conformational exchange on a micro- to millisecond time scale. Binding of a peptide representing the major ELC binding site of the von Hippel-Lindau (VHL) tumor suppressor protein almost completely eliminated chemical exchange processes, but induced substantial conformational changes in Elc1 leading to pronounced rotational anisotropy. These results suggest that elongin C interacts with various target proteins including the VHL protein by an induced fit mechanism involving the conformationally flexible carboxy-terminal helix 4. PMID:10998253

Buchberger, A; Howard, M J; Freund, S M; Proctor, M; Butler, P J; Fersht, A R; Bycroft, M



Metabolic engineering of muconic acid production in Saccharomyces cerevisiae.  


The dicarboxylic acid muconic acid has garnered significant interest due to its potential use as a platform chemical for the production of several valuable consumer bio-plastics including nylon-6,6 and polyurethane (via an adipic acid intermediate) and polyethylene terephthalate (PET) (via a terephthalic acid intermediate). Many process advantages (including lower pH levels) support the production of this molecule in yeast. Here, we present the first heterologous production of muconic acid in the yeast Saccharomyces cerevisiae. A three-step synthetic, composite pathway comprised of the enzymes dehydroshikimate dehydratase from Podospora anserina, protocatechuic acid decarboxylase from Enterobacter cloacae, and catechol 1,2-dioxygenase from Candida albicans was imported into yeast. Further genetic modifications guided by metabolic modeling and feedback inhibition mitigation were introduced to increase precursor availability. Specifically, the knockout of ARO3 and overexpression of a feedback-resistant mutant of aro4 reduced feedback inhibition in the shikimate pathway, and the zwf1 deletion and over-expression of TKL1 increased flux of necessary precursors into the pathway. Further balancing of the heterologous enzyme levels led to a final titer of nearly 141mg/L muconic acid in a shake-flask culture, a value nearly 24-fold higher than the initial strain. Moreover, this strain has the highest titer and second highest yield of any reported shikimate and aromatic amino acid-based molecule in yeast in a simple batch condition. This work collectively demonstrates that yeast has the potential to be a platform for the bioproduction of muconic acid and suggests an area that is ripe for future metabolic engineering efforts. PMID:23164574

Curran, Kathleen A; Leavitt, John M; Karim, Ashty S; Alper, Hal S



Experimental bioenergetics of Saccharomyces cerevisiae in respiration and fermentation  

SciTech Connect

Aerobic growth of Saccharomyces cerevisiae on glucose was investigated, focusing on the heat evolution as it relates to biomass and ethanol synthesis. ''Aerobic fermentation'' and ''aerobic respiration'' were established respectively in the experimental system by performing batch and fed-batch experiments. ''Balanced growth'' batch cultivations were carried out with initial sugar concentrations ranging from 10 to 70 g/L, resulting in different degrees of catabolite repression. The fermentative heat generation was continuously monitored in addition to the key culture parameters such as ethanol production rate, CO2 evolution rate, O2 uptake rate, specific growth rate, and sugar consumption rate. The respective variations of the above quantities reflecting the variations in the catabolic activity of the culture were studied. This was done in order to evaluate the microbial regulatory system, the energetics of microbial growth including the rate of heat evolution and the distribution of organic substrate between respiration and fermentation. This study was supported by closing c, energy, and electron balances on the system. The comparison of the fractions of substrate energy evolved as heat (delta h) with the fraction of available electrons transferred to oxygen (epsilon O2) indicated equal values of the two (0.46) in the aerobic respiration (fed-batch cultivation). However, the glucose effect in batch cultivations resulted in smaller epsilon O2 than delta h, while both values decreased in their absolute values. The evaluation of the heat energetic yield coefficient, together with the fraction of the available electrons transferred to O, contributed to the estimation of the extent of heat production through oxidative phosphorylation. (Refs. 19).

Yerushalmi, L.; Volesky, B.



Experimental bioenergetics of Saccharomyces cerevisiae in respiration and fermentation  

SciTech Connect

Aerobic growth of Saccharomyces cerevisiae on glucose was investigated, focusing on the heat evolution as it relates to biomass and EtOH synthesis. Aerobic fermentation and aerobic respiration were established respectively in the experimental system by performing batch and fed-batch experiments. Balanced growth batch cultivations were carried out with initial sugar concentrations of 10-70 g/L, resulting in different degrees of catabolite repression. The fermentative heat generation was continuously monitored in addition to the key culture parameters such as EtOH production rate, CO/sub 2/ evolution rate, O/sub 2/ uptake rate, sp. growth rate, and sugar consumption rate. The respective variations of the above quantities reflecting the variations in the catabolic activity of the culture were studied. This was done in order to evaluate the microbial regulatory system, the energetics of microbial growth including the rate of heat evolution, and the distribution of organic substrate between respiration and fermentation. This study was supported by closing C, energy, and electron balances on the system. The comparison of the fractions of substrate energy evolved as heat (delta h) with the fraction of available electrons transferred to O/sub 2/ (epsilon O/sub 2/) indicated equal values of the 2 (0.46) in the aerobic respiration (fed-batch cultivation). However, the glucose effect in batch cultivations resulted in smaller epsilon O/sub 2/ than delta h, while both values decreased in their absolute values. The evaluation of the heat energetic yield coefficient, together with the fraction of the available electrons transferred to O/sub 2/, contributed to the estimation of the extent of heat production through oxidative phosphorylation.

Yerushalmi, L.; Volesky, B.



Mutational Analysis of NHAoc/NHA2 in Saccharomyces cerevisiae  

PubMed Central

Background NHAoc/NHA2 is highly and selectively expressed in osteoclasts and plays a role(s) in normal osteoclast differentiation, apoptosis and bone resorptive function in vitro. Extensive mutational analysis of a bacterial homologue, NhaA, has revealed a number of amino acid residues essential for its activity. Some of these residues are evolutionarily conserved and have been shown to be essential not only for activity of NhaA in bacteria, but also of NHAoc/NHA2 in eukaryotes. Methods The salt-sensitive Saccharomyces cerevisiae strain BW31a was used for heterologous expression of mutants of NHAoc/NHA2. Membrane expression of NHAoc/NHA2 was confirmed by confocal microscopy. Intracellular concentration of Na+ (a measure of Na+ antiporter activity) was estimated by atomic absorption spectroscopy. The growth phenotypes of cells expressing NHAoc/NHA2 mutants were studied on YNB agar supplemented with NaCl and by growth curves in YNB broth. Results Mutations in amino acid residues V161 and F357 reduced the ability of transfected BW31a cells to remove intracellular sodium and to grow in NaCl-containing medium. Yeast expressing the double mutant F357 F437 can not grow in 0.4 M NaCl, suggesting that these residues are also essential for antiporter activity. Conclusions Evolutionarily conserved amino acids are required for full antiporter function. General Significance Mutations in these amino acid residues may impact NHAoc activity and therefore osteoclast function in vitro and in vivo.

Huang, Xiaobin; Morse, Leslie R.; Xu, Yan; Zahradka, Jaromir; Sychrova, Hana; Stashenko, Phil; Fan, Feiyue; Battaglino, Ricardo A.



Metabolism and Regulation of Glycerolipids in the Yeast Saccharomyces cerevisiae  

PubMed Central

Due to its genetic tractability and increasing wealth of accessible data, the yeast Saccharomyces cerevisiae is a model system of choice for the study of the genetics, biochemistry, and cell biology of eukaryotic lipid metabolism. Glycerolipids (e.g., phospholipids and triacylglycerol) and their precursors are synthesized and metabolized by enzymes associated with the cytosol and membranous organelles, including endoplasmic reticulum, mitochondria, and lipid droplets. Genetic and biochemical analyses have revealed that glycerolipids play important roles in cell signaling, membrane trafficking, and anchoring of membrane proteins in addition to membrane structure. The expression of glycerolipid enzymes is controlled by a variety of conditions including growth stage and nutrient availability. Much of this regulation occurs at the transcriptional level and involves the Ino2–Ino4 activation complex and the Opi1 repressor, which interacts with Ino2 to attenuate transcriptional activation of UASINO-containing glycerolipid biosynthetic genes. Cellular levels of phosphatidic acid, precursor to all membrane phospholipids and the storage lipid triacylglycerol, regulates transcription of UASINO-containing genes by tethering Opi1 to the nuclear/endoplasmic reticulum membrane and controlling its translocation into the nucleus, a mechanism largely controlled by inositol availability. The transcriptional activator Zap1 controls the expression of some phospholipid synthesis genes in response to zinc availability. Regulatory mechanisms also include control of catalytic activity of glycerolipid enzymes by water-soluble precursors, products and lipids, and covalent modification of phosphorylation, while in vivo function of some enzymes is governed by their subcellular location. Genome-wide genetic analysis indicates coordinate regulation between glycerolipid metabolism and a broad spectrum of metabolic pathways.

Henry, Susan A.; Kohlwein, Sepp D.; Carman, George M.



A vector set for systematic metabolic engineering in Saccharomyces cerevisiae  

PubMed Central

A set of shuttle vectors was constructed to facilitate expression of genes for metabolic engineering in Saccharomyces cerevisiae. Selectable markers include the URA3, TRP1, MET15, LEU2-d8, HIS3 and CAN1 genes. Differential expression of genes can be achieved as each marker is available on both CEN/ARS- and 2 ?-containing plasmids. Unique restriction sites downstream of TEF1, PGK1 or HXT7-391 promoters and upstream of the CYC1 terminator allow insertion of open-reading frame cassettes for expression. Furthermore, a fragment appropriate for integration into the genome via homologous recombination can be readily generated in a polymerase chain reaction. Vector marker genes are flanked by loxP recognition sites for the CreA recombinase to allow efficient site-specific marker deletion and recycling. Expression and copy number were characterized for representative high- and low-copy vectors carrying the different marker and promoter sequences. Metabolic engineering typically requires the stable introduction of multiple genes and genomic integration is often preferred. This requires an expanded number of stable expression sites relative to standard gene expression studies. This study demonstrated the practicality of polymerase chain reaction amplification of an expression cassette and genetic marker, and subsequent replacement of endogenous retrotransposons by homologous recombination with flanking sequences. Such reporters were expressed comparably to those inserted at standard integration loci. This expands the number of available characterized integration sites and demonstrates that such sites provide a virtually inexhaustible pool of integration targets for stable expression of multiple genes. Together these vectors and expression loci will facilitate combinatorial gene expression for metabolic engineering.

Fang, Fang; Salmon, Kirsty; Shen, Michael W. Y.; Aeling, Kimberly A.; Ito, Elaine; Irwin, Becky; Tran, Uyen Phuong C.; Hatfield, G. Wesley; Da Silva, Nancy A.; Sandmeyer, Suzanne



Metabolism and regulation of glycerolipids in the yeast Saccharomyces cerevisiae.  


Due to its genetic tractability and increasing wealth of accessible data, the yeast Saccharomyces cerevisiae is a model system of choice for the study of the genetics, biochemistry, and cell biology of eukaryotic lipid metabolism. Glycerolipids (e.g., phospholipids and triacylglycerol) and their precursors are synthesized and metabolized by enzymes associated with the cytosol and membranous organelles, including endoplasmic reticulum, mitochondria, and lipid droplets. Genetic and biochemical analyses have revealed that glycerolipids play important roles in cell signaling, membrane trafficking, and anchoring of membrane proteins in addition to membrane structure. The expression of glycerolipid enzymes is controlled by a variety of conditions including growth stage and nutrient availability. Much of this regulation occurs at the transcriptional level and involves the Ino2-Ino4 activation complex and the Opi1 repressor, which interacts with Ino2 to attenuate transcriptional activation of UAS(INO)-containing glycerolipid biosynthetic genes. Cellular levels of phosphatidic acid, precursor to all membrane phospholipids and the storage lipid triacylglycerol, regulates transcription of UAS(INO)-containing genes by tethering Opi1 to the nuclear/endoplasmic reticulum membrane and controlling its translocation into the nucleus, a mechanism largely controlled by inositol availability. The transcriptional activator Zap1 controls the expression of some phospholipid synthesis genes in response to zinc availability. Regulatory mechanisms also include control of catalytic activity of glycerolipid enzymes by water-soluble precursors, products and lipids, and covalent modification of phosphorylation, while in vivo function of some enzymes is governed by their subcellular location. Genome-wide genetic analysis indicates coordinate regulation between glycerolipid metabolism and a broad spectrum of metabolic pathways. PMID:22345606

Henry, Susan A; Kohlwein, Sepp D; Carman, George M



Characterization of Glycogen-Deficient Glc Mutants of Saccharomyces Cerevisiae  

PubMed Central

Forty-eight mutants of Saccharomyces cerevisiae with defects in glycogen metabolism were isolated. The mutations defined eight GLC genes, the functions of which were determined. Mutations in three of these genes activate the RAS/cAMP pathway either by impairment of a RAS GTPase-activating protein (GLC1/IRA1 and GLC4/IRA2) or by activating Ras2p (GLC5/RAS2). SNF1 protein kinase (GLC2) was found to be required for normal glycogen levels. Glycogen branching enzyme (GLC3) was found to be required for significant glycogen synthesis. GLC6 was shown to be allelic to CIF1 (and probably FDP1, BYP1 and GGS1), mutations in which were previously found to prevent growth on glucose; this gene is also the same as TPS1, which encodes a subunit of the trehalose-phosphate synthase. Mutations in GLC6 were capable of increasing or decreasing glycogen levels, at least in part via effects on the regulation of glycogen synthase. GLC7 encodes a type 1 protein phosphatase that contributes to the dephosphorylation (and hence activation) of glycogen synthase. GLC8 encodes a homologue of type 1 protein phosphatase inhibitor-2. The genetic map positions of GLC1/IRA1, GLC3, GLC4/IRA2, GLC6/CIF1/TPS1 (and the adjacent VAT2/VMA2), and GLC7 were clarified. From the data on GLC3, there may be a suppression of recombination near the chromosome V centromere, at least in some strains.

Cannon, J. F.; Pringle, J. R.; Fiechter, A.; Khalil, M.



Biogenesis of the Saccharomyces cerevisiae Mating Pheromone a-Factor  

PubMed Central

The Saccharomyces cerevisiae mating pheromone a-factor is a prenylated and carboxyl methylated extracellular peptide signaling molecule. Biogenesis of the a-factor precursor proceeds via a distinctive multistep pathway that involves COOH-terminal modification, NH2-terminal proteolysis, and a nonclassical export mechanism. In this study, we examine the formation and fate of a-factor biosynthetic intermediates to more precisely define the events that occur during a-factor biogenesis. We have identified four distinct a-factor biosynthetic intermediates (P0, P1, P2, and M) by metabolic labeling, immunoprecipitation, and SDSPAGE. We determined the biochemical composition of each by defining their NH2-terminal amino acid and COOH-terminal modification status. Unexpectedly, we discovered that not one, but two NH2-terminal cleavage steps occur during the biogenesis of a-factor. In addition, we have shown that COOH-terminal prenylation is required for the NH2-terminal processing of a-factor and that all the prenylated a-factor intermediates (P1, P2, and M) are membrane bound, suggesting that many steps of a-factor biogenesis occur in association with membranes. We also observed that although the biogenesis of a-factor is a rapid process, it is inherently inefficient, perhaps reflecting the potential for regulation. Previous studies have identified gene products that participate in the COOH-terminal modification (Ram1p, Ram2p, Ste14p), NH2-terminal processing (Ste24p, Axl1p), and export (Ste6p) of a-factor. The intermediates defined in the present study are discussed in the context of these biogenesis components to formulate an overall model for the pathway of a-factor biogenesis.

Chen, Peng; Sapperstein, Stephanie K.; Choi, Jonathan D.; Michaelis, Susan



Control of meiotic gene expression in Saccharomyces cerevisiae.  

PubMed Central

Sporulation of the yeast Saccharomyces cerevisiae is restricted to one type of cell, the a/alpha cell, and is initiated after starvation for nitrogen in the absence of a fermentable carbon source. More than 25 characterized genes are expressed only during sporulation and are referred to as meiotic genes or sporulation-specific genes. These genes are in the early, middle, and late expression classes. Most early genes have a 5' regulatory site, URS1, and one of two additional sequences, UASH or a T4C site. URS1 is required both to repress meiotic genes during vegetative growth and to activate these genes during meiosis. UASH and the T4C site also contribute to meiotic expression. A different type of site, the NRE, is found in at least two late genes. The NRE behaves as a repression site in vegetative cells and is neutral in meiotic cells. Many regulatory genes that either repress or activate meiotic genes have been identified. One group of regulators affects the expression of IME1, which specifies a positive regulator of meiotic genes and is expressed at the highest levels in meiotic cells. A second group of regulators acts in parallel with or downstream of IME1 to influence meiotic gene expression. This group includes UME6, which is required both for repression through the URS1 site in vegetative cells and for IME1-dependent activation of an upstream region containing URS1 and T4C sites. IME1 may activate meiotic genes by modifying a UME6-dependent repression complex at a URS1 site. Several additional mechanisms restrict functional expression of some genes to meiotic cells. Translation of IME1 has been proposed to occur only in meiotic cells; several meiotic transcripts are more stable in acetate medium than in glucose medium; and splicing of MER2 RNA depends on a meiosis-specific gene, MER1.

Mitchell, A P



Complete kinetic mechanism of homoisocitrate dehydrogenase from Saccharomyces cerevisiae.  


The kinetic mechanism of homoisocitrate dehydrogenase from Saccharomyces cerevisiae was determined using initial velocity studies in the absence and presence of product and dead end inhibitors in both reaction directions. Data suggest a steady state random kinetic mechanism. The dissociation constant of the Mg-homoisocitrate complex (MgHIc) was estimated to be 11 +/- 2 mM as measured using Mg2+ as a shift reagent. Initial velocity data indicate the MgHIc complex is the reactant in the direction of oxidative decarboxylation, while in the reverse reaction direction, the enzyme likely binds uncomplexed Mg2+ and alpha-ketoadipate. Curvature is observed in the double-reciprocal plots for product inhibition by NADH and the dead-end inhibition by 3-acetylpyridine adenine dinucleotide phosphate when MgHIc is the varied substrate. At low concentrations of MgHIc, the inhibition by both nucleotides is competitive, but as the MgHIc concentration increases, the inhibition changes to uncompetitive, consistent with a steady state random mechanism with preferred binding of MgHIc before NAD. Release of product is preferred and ordered with respect to CO2, alpha-ketoadipate, and NADH. Isocitrate is a slow substrate with a rate (V/E(t)) 216-fold slower than that measured with HIc. In contrast to HIc, the uncomplexed form of isocitrate and Mg2+ bind to the enzyme. The kinetic mechanism in the direction of oxidative decarboxylation of isocitrate, on the basis of initial velocity studies in the absence and presence of dead-end inhibitors, suggests random addition of NAD and isocitrate with Mg2+ binding before isocitrate in rapid equilibrium, and the mechanism approximates rapid equilibrium random. The Keq for the overall reaction measured directly using the change in NADH as a probe is 0.45 M. PMID:17223711

Lin, Ying; Alguindigue, Susan S; Volkman, Jerome; Nicholas, Kenneth M; West, Ann H; Cook, Paul F



Metabolic engineering for enhanced fatty acids synthesis in Saccharomyces cerevisiae.  


Microbial production of biofuel has attracted significant attention in recent years. The fatty acids are important precursors for the production of fuels and chemicals, and its biosynthesis is initiated by the conversion of acetyl-CoA to malonyl-CoA which requires acetyl-CoA as key substrate. Herein, the yeast Saccharomyces cerevisiae was proposed to be metabolically engineered for cytosol acetyl-CoA enhancement for fatty acid synthesis. By gene disruption strategy, idh1 and idh2 genes involved in citrate turnover in tricarboxylic acid cycle (TCA cycle) were disrupted and the citrate production level was increased to 4- and 5-times in mutant yeast strains. In order to convert accumulated citrate to cytosol acetyl-CoA, a heterologous ATP-citrate lyase (ACL) was overexpressed in yeast wild type and idh1,2 disrupted strains. The wild type strain expressing acl mainly accumulated saturated fatty acids: C14:0, C16:0 and C18:0 at levels about 20%, 14% and 27%, respectively. Additionally, the idh1,2 disrupted strains expressing acl mainly accumulated unsaturated fatty acids. Specifically in ?idh1 strain expressing acl, 80% increase in C16:1 and 60% increase in C18:1 was detected. In ?idh2 strain expressing acl, 60% increase in C16:1 and 45% increase in C18:1 was detected. In ?idh1/2 strain expressing acl, there was 92% increase in C16:1 and 77% increase in C18:1, respectively. The increased fatty acids from our study may well be potential substrates for the production of hydrocarbon molecules as potential biofuels. PMID:23353549

Tang, Xiaoling; Feng, Huixing; Chen, Wei Ning



Fermentation performance and intracellular metabolite patterns in laboratory and industrial xylose-fermenting Saccharomyces cerevisiae  

Microsoft Academic Search

.   Heterologous genes for xylose utilization were introduced into an industrial Saccharomyces cerevisiae, strain A, with the aim of producing fuel ethanol from lignocellulosic feedstocks. Two transformants, A4 and A6, were evaluated\\u000a by comparing the performance in 4-l anaerobic batch cultivations to both the parent strain and a laboratory xylose-utilizing\\u000a strain: S. cerevisiae TMB 3001. During growth in a minimal

J. Zaldivar; A. Borges; B. Johansson; H. Smits; S. Villas-Bôas; J. Nielsen; L. Olsson



Repeated-batch production of glucoamylase using recombinant Saccharomyces cerevisiae immobilized in a fibrous bed bioreactor  

Microsoft Academic Search

The recombinant Saccharomyces cerevisiae strain C468\\/pGAC9 has an unstable hybrid plasmid pGAC9, which directs production of glucoamylase. A fibrous cotton material\\u000a with a good adsorption capability for recombinant S. cerevisiae cells was used as the immobilization matrix in an internal loop airlift-driven fibrous bed bioreactor (ILALFBB) system. With\\u000a batch cultures in the ILALFBB, the fraction of plasmid-carrying cells was 72%

Peter M. Kilonzo; Argyrios Margaritis; Maurice A. Bergougnou



Anaerobic respiration of superoxide dismutase-deficient Saccharomyces cerevisiae under oxidative stress  

Microsoft Academic Search

The ethanol productivity of superoxide dismutase (SOD)-deficient mutants ofSaccharomyces cerevisiae was examined under the oxidative stress by Paraquat. It was observed that MnSOD-deficient mutant ofS. cerevisiae had higher ethanol productivity than wild type or CuZnSOD-deficient yeast both in aerobic and in anaerobic culture condition.\\u000a Pyruvate dehydrogenase activity decreased by 35% and alcohol dehydrogenase activity increased by 32% were observed in

Sun Mi Lee; Doo Hyun Nam



Identification of Gal80p-interacting proteins by Saccharomyces cerevisiae whole genome phage display  

Microsoft Academic Search

Networks of interacting proteins and protein interaction maps can help in functional annotation in genome analysis projects. We present the application of genomic phage display as a tool to identify interacting proteins in Saccharomyces cerevisiae. We have developed a large phagemid display library (?7.7×107 independent clones) of sheared S. cerevisiae genomic DNA (12.1 Mbp genome size) fused to gene III

Kirsten Hertveldt; Mekonnen Lemma Dechassa; Johan Robben; Guido Volckaert



Inactivation of Saccharomyces cerevisiae and Bacillus cereus by pulsed electric fields technology  

Microsoft Academic Search

The effect of pulsed electric field (PEF) treatment, applied in a continuous system, on Saccharomyces cerevisiae and Bacillus cereus cells and spores was investigated. S. cerevisiae inoculated into sterilised apple juice and B. cereus cells and spores inoculated into sterilised 0.15% NaCl were treated with electric field strengths of 10–28 kV\\/cm using an 8.3 pulse number and with pulse numbers

Zs. Cserhalmi; I. Vidács; J. Beczner; B. Czukor



Genome-wide transcriptional analysis of Saccharomyces cerevisiae during industrial bioethanol fermentation  

Microsoft Academic Search

Saccharomyces cerevisiae is widely applied in large-scale industrial bioethanol fermentation; however, little is known about the molecular responses\\u000a of industrial yeast during large-scale fermentation processes. We investigated the global transcriptional responses of an\\u000a industrial strain of S. cerevisiae during industrial continuous and fed-batch fermentation by oligonucleotide-based microarrays. About 28 and 62% of all genes\\u000a detected showed differential gene expression during

Bing-Zhi Li; Jing-Sheng Cheng; Bin Qiao; Ying-Jin Yuan



Use of Saccharomyces cerevisiae BLYES expressing bacterial bioluminescence for rapid, sensitive detection of estrogenic compounds  

Microsoft Academic Search

An estrogen-inducible bacterial lux-based bioluminescent reporter was developed in Saccharomyces cerevisiae for applications in chemical sensing and environmental assessment of estrogen disruptor activity. The strain, designated S. cerevisiae BLYES, was constructed by inserting tandem estrogen response elements between divergent yeast promoters GPD and ADH1 on pUTK401 (formerly pUA12B7) that constitutively express luxA and luxB to create pUTK407. Cotransformation of this

John Sanseverino; Rakesh K. Gupta; Alice C. Layton; Stacey S. Patterson; Steven A. Ripp; Leslie Saidak; Michael L. Simpson; T. Wayne Schultz; Gary S. Sayler



Protein Kinase A, TOR, and Glucose Transport Control the Response to Nutrient Repletion in Saccharomyces cerevisiae  

Microsoft Academic Search

Nutrient repletion leads to substantial restructuring of the transcriptome in Saccharomyces cerevisiae. The expression levels of approximately one-third of all S. cerevisiae genes are altered at least twofold when a nutrient-depleted culture is transferred to fresh medium. Several nutrient-sensing pathways are known to play a role in this process, but the relative contribution that each pathway makes to the total

Matthew G. Slattery; Dritan Liko; Warren Heideman



Development and characterisation of a recombinant Saccharomyces cerevisiae mutant strain with enhanced xylose fermentation properties  

Microsoft Academic Search

The purpose of this study was to help lay the foundation for further development of xylose-fermentingSaccharomyces cerevisiae yeast strains through an approach that combined metabolic engineering and random mutagenesis in a recombinant haploid strain\\u000a that overexpressed only two genes of the xylose pathway. Previously,S. cerevisiae strains, overexpressing heterologous genes encoding xylose reductase, xylitol dehydrogenase and the endogenousXKS1 xylulokinase gene, were

Vasudevan Thanvanthri Gururajan; Pierre VAN RENSBURG; Bärbel Hahn-Hägerdal; Isak S. Pretorius; Ricardo R. Cordero Otero



DNA sequence analysis of spontaneous mutations in the SUP4-o gene of Saccharomyces cerevisiae.  

PubMed Central

A collection of 196 spontaneous mutations in the SUP4-o gene of the yeast Saccharomyces cerevisiae was analyzed by DNA sequencing. The classes of mutation identified included all possible types of base-pair substitution, deletions of various lengths, complex alterations involving multiple changes, and insertions of transposable elements. Our findings demonstrate that at least several different mechanisms are responsible for spontaneous mutagenesis in S. cerevisiae.

Giroux, C N; Mis, J R; Pierce, M K; Kohalmi, S E; Kunz, B A



Evaluation of wet-feeding wheat-based diets containing Saccharomyces cerevisiae to broiler chickens  

Microsoft Academic Search

1. This experiment investigated the effects of water and Saccharomyces cerevisiae added to wheat-based diets on gastrointestinal, blood and performance parameters of broiler chickens.2. A total of 160 one-d-old male broiler chicks were given air-dry or wet diets, with or without S. cerevisiae supplementation (0 and 20?g\\/kg air-dry feed) ad libitum to 42?d.3. Feeding broilers with a diet mixed with

M. Afsharmanesh; M. Barani; F. G. Silversides



Computational identification of non-coding RNAs in Saccharomyces cerevisiae by comparative genomics  

Microsoft Academic Search

We screened for new structural non-coding RNAs (ncRNAs) in the genome sequence of the yeast Saccharomyces cerevisiae using computational comparative analysis of genome sequences from five related species of Saccharomyces. The screen identified 92 candidate ncRNA genes. Thirteen showed discrete transcripts when assayed by northern blot. Of these, eight appear to be novel ncRNAs ranging in size from 268 to

John P. McCutcheon; Sean R. Eddy



Maize Activator (Ac) Transposase (TPase) Is Expressed in Saccharomyces cerevisiae from a Genomic Clone  

Microsoft Academic Search

The maize Activator (Ac) transposase (TPase) was expressed as a Histidine (His)-tagged protein in Saccharomyces cerevisiae from a full length genomic clone. Expression was demonstrated via the highly specific nickel-coated Elisa plate method, using an anti-His antibody and 2 separate anti-Ac TPase antibodies, to Ac residues 103–465 and 189–807. Ac TPase expression in Saccharomyces is important for two reasons: (a)

Amy F. MacRae



Controlled formation of volatile components in cider making using a combination of Saccharomyces cerevisiae and Hanseniaspora valbyensis yeast species  

Microsoft Academic Search

The effect of pure and mixed fermentation by Saccharomyces cerevisiae and Hanseniaspora valbyensis on the formation of major volatile components in cider was investigated. When the interaction between yeast strains of S. cerevisiae and H. valbyensis was studied, it was found that the two strains each affected the cell growth of the other upon inoculation of S. cerevisiae during growth

Y. Xu; G. A. Zhao; L. P. Wang



Intracellular Maltose Is Sufficient To Induce MAL Gene Expression in Saccharomyces cerevisiae  

PubMed Central

The presence of maltose induces MAL gene expression in Saccharomyces cells, but little is known about how maltose is sensed. Strains with all maltose permease genes deleted are unable to induce MAL gene expression. In this study, we examined the role of maltose permease in maltose sensing by substituting a heterologous transporter for the native maltose permease. PmSUC2 encodes a sucrose transporter from the dicot plant Plantago major that exhibits no significant sequence homology to maltose permease. When expressed in Saccharomyces cerevisiae, PmSUC2 is capable of transporting maltose, albeit at a reduced rate. We showed that introduction of PmSUC2 restores maltose-inducible MAL gene expression to a maltose permease-null mutant and that this induction requires the MAL activator. These data indicate that intracellular maltose is sufficient to induce MAL gene expression independently of the mechanism of maltose transport. By using strains expressing defective mal61 mutant alleles, we demonstrated a correlation between the rate of maltose transport and the level of the induction, which is particularly evident in medium containing very limiting concentrations of maltose. Moreover, our results indicate that a rather low concentration of intracellular maltose is needed to trigger MAL gene expression. We also showed that constitutive overexpression of either MAL61 maltose permease or PmSUC2 suppresses the noninducible phenotype of a defective mal13 MAL-activator allele, suggesting that this suppression is solely a function of maltose transport activity and is not specific to the sequence of the permease. Our studies indicate that maltose permease does not function as the maltose sensor in S. cerevisiae.

Wang, Xin; Bali, Mehtap; Medintz, Igor; Michels, Corinne A.



Novel role for a Saccharomyces cerevisiae nucleoporin, Nup170p, in chromosome segregation.  

PubMed Central

We determined that a mutation in the nucleoporin gene NUP170 leads to defects in chromosome transmission fidelity (ctf) and kinetochore integrity in Saccharomyces cerevisiae. A ctf mutant strain, termed s141, shows a transcription readthrough phenotype and stabilizes a dicentric chromosome fragment in two assays for kinetochore integrity. Previously, these assays led to the identification of two essential kinetochore components, Ctf13p and Ctf14p. Thus, s141 represents another ctf mutant involved in the maintenance of kinetochore integrity. We cloned and mapped the gene complementing the ctf mutation of s141 and showed that it is identical to the S. cerevisiae NUP170 gene. A deletion strain of NUP170 (nup170 Delta::HIS3) has a Ctf(-) phenotype similar to the s141 mutant (nup170-141) and also exhibits a kinetochore integrity defect. We identified a second nucleoporin, NUP157, a homologue of NUP170, as a suppressor of the Ctf(-) phenotype of nup170-141 and nup170 Delta::HIS3 strains. However, a deletion of NUP157 or several other nucleoporins did not affect chromosome segregation. Our data suggest that NUP170 encodes a specialized nucleoporin with a unique role in chromosome segregation and possibly kinetochore function.

Kerscher, O; Hieter, P; Winey, M; Basrai, M A



Acquisition of thermotolerant yeast Saccharomyces cerevisiae by breeding via stepwise adaptation.  


A thermotolerant Saccharomyces cerevisiae yeast strain, YK60-1, was bred from a parental strain, MT8-1, via stepwise adaptation. YK60-1 grew at 40(o) C, a temperature at which MT8-1 could not grow at all. YK60-1 exhibited faster growth than MT8-1 at 30(o) C. To investigate the mechanisms how MT8-1 acquired thermotolerance, DNA microarray analysis was performed. The analysis revealed that induction of stress-responsive genes such as those encoding heat shock proteins and trehalose biosynthetic enzymes in YK60-1. Furthermore, non-targeting metabolome analysis showed that YK60-1 accumulated more trehalose, a metabolite that contributes to stress tolerance in yeast, than MT8-1. In conclusion, S. cerevisiae MT8-1 acquired thermotolerance by induction of specific stress-responsive genes and enhanced intracellular trehalose levels. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 2013. PMID:23681509

Satomura, Atsushi; Katsuyama, Yoshiaki; Miura, Natsuko; Kuroda, Kouichi; Tomio, Ayako; Bamba, Takeshi; Fukusaki, Eiichiro; Ueda, Mitsuyoshi



Involvement of mitochondria in the assimilatory metabolism of anaerobic Saccharomyces cerevisiae cultures.  


The possible physiological role of mitochondria in anaerobically grown Saccharomyces cerevisiae was investigated via enzyme localization and inhibitor studies. Almost all of the activity of citrate synthase (EC was recovered in the mitochondrial fraction after differential centrifugation of spheroplast lysates. The enzyme exhibited a high degree of latency which was demonstrated by sonication of the mitochondrial fractions. Since citrate synthase is an important enzyme in anabolic reactions, a consequence of this localization is the requirement for transport of metabolites across the mitochondrial membranes. Such transport is likely to require energy which, as a result of anaerobiosis, cannot be supplied by respiration. It was therefore investigated whether ATP translocation into the mitochondria by an ADP/ATP translocase might be involved in anaerobic mitochondrial energy metabolism. It was shown that addition of the ADP/ATP translocase inhibitor bongkrekic acid to anaerobic cultures indeed inhibited growth, although only partially. It is concluded that mitochondria of S. cerevisiae fulfil a vital role in anaerobic sugar metabolism. PMID:7812444

Visser, W; van der Baan, A A; Batenburg-van der Vegte, W; Scheffers, W A; Krämer, R; van Dijken, J P



Alcoholic fermentation of xylose and mixed sugars using recombinant Saccharomyces cerevisiae engineered for xylose utilization.  


Previously, a Saccharomyces cerevisiae strain was engineered for xylose assimilation by the constitutive overexpression of the Orpinomyces xylose isomerase, the S. cerevisiae xylulokinase, and the Pichia stipitis SUT1 sugar transporter genes. The recombinant strain exhibited growth on xylose, under aerobic conditions, with a specific growth rate of 0.025 h(-1), while ethanol production from xylose was achieved anaerobically. In the present study, the developed recombinant yeast was adapted for enhanced growth on xylose by serial transfer in xylose-containing minimal medium under aerobic conditions. After repeated batch cultivations, a strain was isolated which grew with a specific growth rate of 0.133 h(-1). The adapted strain could ferment 20 g l(-1) of xylose to ethanol with a yield of 0.37 g g(-1) and production rate of 0.026 g l(-1) h(-1). Raising the fermentation temperature from 30 degrees C to 35 degrees C resulted in a substantial increase in the ethanol yield (0.43 g g(-1)) and production rate (0.07 g l(-1) h(-1)) as well as a significant reduction in the xylitol yield. By the addition of a sugar complexing agent, such as sodium tetraborate, significant improvement in ethanol production and reduction in xylitol accumulation was achieved. Furthermore, ethanol production from xylose and a mixture of glucose and xylose was also demonstrated in complex medium containing yeast extract, peptone, and borate with a considerably high yield of 0.48 g g(-1). PMID:19125247

Madhavan, Anjali; Tamalampudi, Sriappareddy; Srivastava, Aradhana; Fukuda, Hideki; Bisaria, Virendra S; Kondo, Akihiko



Respiratory capacities of mitochondria of Saccharomyces cerevisiae CBS 8066 and Candida utilis CBS 621 grown under glucose limitation.  


A comparative study was made of the in vitro respiratory capacity of mitochondria isolated from Saccharomyces cerevisiae and Candida utilis grown in glucose-limited chemostat cultures. An electron-microscopic analysis of whole cells revealed that the volume density of mitochondria was the same in both yeasts. Mitochondria from both organisms exhibited respiratory control with NADH, pyruvate + malate, 2-oxoglutarate + acetate or malate, and ethanol. The rate of oxidation of these compounds by isolated mitochondria was the same in both yeasts. The rate of oxidation of NADPH by mitochondria from S. cerevisiae was 10 times lower than by those from C. utilis. However, this low rate probably has no influence on the overall in vivo respiratory capacity of S. cerevisiae. The results are discussed in relation to the differences in metabolic behaviour between S. cerevisiae and C. utilis upon transition of cultures from glucose limitation to glucose excess. It is concluded that the occurrence of alcoholic fermentation in S. cerevisiae under these conditions does not result from a bottleneck in the respiratory capacity of the mitochondria. PMID:2686549

Van Urk, H; Bruinenberg, P M; Veenhuis, M; Scheffers, W A; Van Dijken, J P



Key cytomembrane ABC transporters of Saccharomyces cerevisiae fail to improve the tolerance to D-limonene.  


ATP-binding cassette transporters (ABC) are important detoxification proteins and were proposed to play important roles in monoterpene resistance in Saccharomyces cerevisiae. In this work, the transcriptional levels of typical ABC transporters of S. cerevisiae under 85 mg D-limonene/l were evaluated using real-time quantitative PCR. Only the transcriptional level of PDR5, YOR1 and PDR15 were upregulated but overexpression of these genes in S. cerevisiae failed to improve D-limonene tolerance suggesting that other mechanisms are involved in tolerance of yeast to monoterpenes. PMID:22526424

Hu, Feifei; Liu, Jidong; Du, Guocheng; Hua, Zhaozhe; Zhou, Jingwen; Chen, Jian



Genome Snapshot: a new resource at the Saccharomyces Genome Database (SGD) presenting an overview of the Saccharomyces cerevisiae genome  

PubMed Central

Sequencing and annotation of the entire Saccharomyces cerevisiae genome has made it possible to gain a genome-wide perspective on yeast genes and gene products. To make this information available on an ongoing basis, the Saccharomyces Genome Database (SGD) () has created the Genome Snapshot (). The Genome Snapshot summarizes the current state of knowledge about the genes and chromosomal features of S.cerevisiae. The information is organized into two categories: (i) number of each type of chromosomal feature annotated in the genome and (ii) number and distribution of genes annotated to Gene Ontology terms. Detailed lists are accessible through SGD's Advanced Search tool (), and all the data presented on this page are available from the SGD ftp site ().

Hirschman, Jodi E.; Balakrishnan, Rama; Christie, Karen R.; Costanzo, Maria C.; Dwight, Selina S.; Engel, Stacia R.; Fisk, Dianna G.; Hong, Eurie L.; Livstone, Michael S.; Nash, Robert; Park, Julie; Oughtred, Rose; Skrzypek, Marek; Starr, Barry; Theesfeld, Chandra L.; Williams, Jennifer; Andrada, Rey; Binkley, Gail; Dong, Qing; Lane, Christopher; Miyasato, Stuart; Sethuraman, Anand; Schroeder, Mark; Thanawala, Mayank K.; Weng, Shuai; Dolinski, Kara; Botstein, David; Cherry, J. Michael



Substrate Specificity of Thiamine Pyrophosphate-Dependent 2-Oxo-Acid Decarboxylases in Saccharomyces cerevisiae  

PubMed Central

Fusel alcohols are precursors and contributors to flavor and aroma compounds in fermented beverages, and some are under investigation as biofuels. The decarboxylation of 2-oxo acids is a key step in the Ehrlich pathway for fusel alcohol production. In Saccharomyces cerevisiae, five genes share sequence similarity with genes encoding thiamine pyrophosphate-dependent 2-oxo-acid decarboxylases (2ODCs). PDC1, PDC5, and PDC6 encode differentially regulated pyruvate decarboxylase isoenzymes; ARO10 encodes a 2-oxo-acid decarboxylase with broad substrate specificity, and THI3 has not yet been shown to encode an active decarboxylase. Despite the importance of fusel alcohol production in S. cerevisiae, the substrate specificities of these five 2ODCs have not been systematically compared. When the five 2ODCs were individually overexpressed in a pdc1? pdc5? pdc6? aro10? thi3? strain, only Pdc1, Pdc5, and Pdc6 catalyzed the decarboxylation of the linear-chain 2-oxo acids pyruvate, 2-oxo-butanoate, and 2-oxo-pentanoate in cell extracts. The presence of a Pdc isoenzyme was also required for the production of n-propanol and n-butanol in cultures grown on threonine and norvaline, respectively, as nitrogen sources. These results demonstrate the importance of pyruvate decarboxylases in the natural production of n-propanol and n-butanol by S. cerevisiae. No decarboxylation activity was found for Thi3 with any of the substrates tested. Only Aro10 and Pdc5 catalyzed the decarboxylation of the aromatic substrate phenylpyruvate, with Aro10 showing superior kinetic properties. Aro10, Pdc1, Pdc5, and Pdc6 exhibited activity with all branched-chain and sulfur-containing 2-oxo acids tested but with markedly different decarboxylation kinetics. The high affinity of Aro10 identified it as a key contributor to the production of branched-chain and sulfur-containing fusel alcohols.

Romagnoli, Gabriele; Luttik, Marijke A. H.; Kotter, Peter; Pronk, Jack T.



Nanoscale effects of caspofungin against two yeast species, Saccharomyces cerevisiae and Candida albicans.  


Saccharomyces cerevisiae and Candida albicans are model yeasts for biotechnology and human health, respectively. We used atomic force microscopy (AFM) to explore the effects of caspofungin, an antifungal drug used in hospitals, on these two species. Our nanoscale investigation revealed similar, but also different, behaviors of the two yeasts in response to treatment with the drug. While administration of caspofungin induced deep cell wall remodeling in both yeast species, as evidenced by a dramatic increase in chitin and decrease in ?-glucan content, changes in cell wall composition were more pronounced with C. albicans cells. Notably, the increase of chitin was proportional to the increase in the caspofungin dose. In addition, the Young modulus of the cell was three times lower for C. albicans cells than for S. cerevisiae cells and increased proportionally with the increase of chitin, suggesting differences in the molecular organization of the cell wall between the two yeast species. Also, at a low dose of caspofungin (i.e., 0.5× MIC), the cell surface of C. albicans exhibited a morphology that was reminiscent of cells expressing adhesion proteins. Interestingly, this morphology was lost at high doses of the drug (i.e., 4× MIC). However, the treatment of S. cerevisiae cells with high doses of caspofungin resulted in impairment of cytokinesis. Altogether, the use of AFM for investigating the effects of antifungal drugs is relevant in nanomedicine, as it should help in understanding their mechanisms of action on fungal cells, as well as unraveling unexpected effects on cell division and fungal adhesion. PMID:23669379

Formosa, C; Schiavone, M; Martin-Yken, H; François, J M; Duval, R E; Dague, E



Growth and Fermentation Characteristics of Saccharomyces cerevisiae NK28 Isolated from Kiwi Fruit.  


The influences of glucose concentration, initial medium acidity (pH), and temperature on the growth and ethanol production of Saccharomyces cerevisiae NK28, which was isolated from kiwi fruit, were examined in shake flask cultures. The optimal glucose concentration, initial medium pH, and temperature for ethanol production were 200 g/l, pH 6.0, and 35oC, respectively. Under this growth condition, S. cerevisiae NK28 produced 98.9 ± 5.67 g/l ethanol in 24 h with a volumetric ethanol production rate of 4.12 ± 0.24 g/l·h. S. cerevisiae NK28 was more tolerant to heat and ethanol than laboratory strain S. cerevisiae BY4742, and its tolerance to ethanol and fermentation inhibitors was comparable to that of an ethanologen, S. cerevisiae D5A. PMID:23893096

Lee, Jong-Sub; Park, Eun-Hee; Kim, Jung-Wan; Yeo, Soo-Hwan; Kim, Myoung-Dong



Clinical Saccharomyces cerevisiae isolates cannot cross the epithelial barrier in vitro.  


Saccharomyces cerevisiae is generally considered to be a safe organism and is essential to produce many different kinds of foods as well as being widely used as a dietary supplement. However, several isolates, which are genetically related to brewing and baking yeasts, have shown virulent traits, being able to produce human infections in immunodeficient patients. Previously it has been shown that the administration of S. cerevisiae clinical isolates can lead to systemic infections, reaching several organs in murine systems. In this work, we studied S. cerevisiae clinical isolates in an in vitro intestinal epithelial barrier model, comparing their behaviour with that of several strains of the related pathogens Candida glabrata and Candida albicans. The results showed that, in contrast to C. glabrata and C. albicans, S. cerevisiae was not able to cross the intestinal barrier. We concluded that S. cerevisiae can only perform opportunistic or passive crossings when epithelial barrier integrity is previously compromised. PMID:22609000

Pérez-Torrado, Roberto; Llopis, Silvia; Jespersen, Lene; Fernández-Espinar, Teresa; Querol, Amparo



Fsy1, the sole hexose-proton transporter characterized in Saccharomyces yeasts, exhibits a variable fructose:H(+) stoichiometry.  


In the model yeast Saccharomyces cerevisiae, hexose uptake is mediated exclusively by a family of facilitators (Hxt, hexose transporters). Some other Saccharomyces species (e.g. Saccharomyces bayanus and Saccharomyces pastorianus) possess, in addition, a specific fructose transporter (Fsy1, fructose symporter) that has been previously described to function as a proton symporter. In the present work, we compared growth of a yeast strain in which FSY1 occurs naturally in anaerobic, fructose- and glucose-limited chemostat cultures. Especially at low specific growth rates, fructose-proton symport was shown to have a strong impact on the biomass yield on sugar. We subsequently employed energized hybrid plasma membrane vesicles to confirm previous observations concerning the mode of operation and specificity of Fsy1 mediated transport. Surprisingly, these experiments suggested that the carrier exhibits an unusual fructose:H(+) stoichiometry of 1:2. This energetically expensive mode of operation was also found consistently in vivo, in shake flask and in chemostat cultures, and both when Fsy1 is the sole transporter and when the Hxt carriers are present. However, it is observed only when Fsy1 is operating at higher glycolytic fluxes, a situation that is normally prevented by downregulation of the gene. Taken together, our results suggest the possibility that fructose symport with more than one proton may constitute an energetically unfavorable mode of operation of the Fsy1 transporter that, in growing cultures, is prevented by transcriptional regulation. PMID:22922355

Anjos, Jorge; Rodrigues de Sousa, Helena; Roca, Christophe; Cássio, Fernanda; Luttik, Marijke; Pronk, Jack T; Salema-Oom, Madalena; Gonçalves, Paula



Structural and Kinetic Isotope Effect Studies of Nicotinamidase (Pnc1) from Saccharomyces cerevisiae  

Microsoft Academic Search

Nicotinamidases catalyze the hydrolysis of nicotinamide to nicotinic acid and ammonia. Nicotinamidases are absent in mammals but function in NAD{sup +} salvage in many bacteria, yeast, plants, protozoa, and metazoans. We have performed structural and kinetic investigations of the nicotinamidase from Saccharomyces cerevisiae (Pnc1). Steady-state product inhibitor analysis revealed an irreversible reaction in which ammonia is the first product released,

Brian C. Smith; Mark A. Anderson; Kelly A. Hoadley; James L. Keck; W. Wallace Cleland; John M. Denu



Aminopeptidase I of Saccharomyces cerevisiae is localized to the vacuole independent of the secretory pathway  

Microsoft Academic Search

The Saccharomyces cerevisiae APE1 gene product, aminopeptidase I (API), is a soluble hydro- lase that has been shown to be localized to the vac- uole. API lacks a standard signal sequence and con- tains an unusual amino-terminal propeptide. We have examined the biosynthesis of API in order to elucidate the mechanism of its delivery to the vacuole. API is synthesized

Daniel J. Klionsky; Rosario Cueva; Debbie S. Yaver



The wheat LEA protein Em functions as an osmoprotective molecule in Saccharomyces cerevisiae  

Microsoft Academic Search

The biased amino acid composition and aperiodic (random coil) configuration of Group 1 late embryogenesis-abundant (LEA) proteins imply that these proteins are capable of binding large amounts of water. While Group 1 LEAs have been predicted to contribute to osmotic stress protection in both embryonic and vegetative tissues, biochemical support has been lacking. We have used Saccharomyces cerevisiae as a

Ginger A. Swire-Clark; William R. Marcotte



11 Integration of nutrient signalling pathways in the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

The ability to sense and to respond to changes in the nutrient availability is an essential feature for the survival of every organism. Saccharomyces cerevisiae has several signal transduction cascades to optimally adapt its metabolism to the availability of nutrients in the environment. In this chapter, we focus on the convergence of signal transduction pathways for nutrient sensing in budding

Johnny Roosen; Christine Oesterhelt; Katrien Pardons; Erwin Swinnen; Joris Winderickx


Comparative Analysis of the Saccharomyces cerevisiae and Caenorhabditis elegans Protein Interaction Network  

Microsoft Academic Search

Protein interaction networks aim to summarize the complex interplay of proteins in an organism. Early studies suggested that the position of a protein in the network determines its evolutionary rate but there has been considerable disagreement as to what extent other factors, such as protein abundance, modify this reported dependence. We compare the genomes of Saccharomyces cerevisiae and Caenorhabditis elegans

Ino Agrafioti; Jonathan Swire; James Abbott; Derek Huntley; Sarah Butcher; Michael P. H. Stumpf



Stress Tolerance in Doughs of Saccharomyces cerevisiae Trehalase Mutants Derived from Commercial Baker's Yeast  

Microsoft Academic Search

Accumulation of trehalose is widely believed to be a critical determinant in improving the stress tolerance of the yeast Saccharomyces cerevisiae, which is commonly used in commercial bread dough. To retain the accumulation of trehalose in yeast cells, we constructed, for the first time, diploid homozygous neutral trehalase mutants (Dnth1), acid trehalase mutants (Dath1), and double mutants (Dnth1 ath1) by




Identification of Candida albicans Genes that Induce Saccharomyces cerevisiae Cell Adhesion and Morphogenesis  

Microsoft Academic Search

Morphogenesis and adhesion to host tissues and medical devices contribute to the virulence of Candida albicans, the most common fungal pathogen isolated from humans. However, identification of molecular mechanisms of C. albicans adhesion and morphogenesis has been impaired by the lack of effective molecular and genetic tools available for this organism. Saccharomyces cerevisiae provides an attractive model system for studying

Fang Li; Sean P. Palecek




Technology Transfer Automated Retrieval System (TEKTRAN)

The gene encoding Lentinula edodes glucoamylase (GLA) was cloned into Saccharomyces cerevisiae, expressed constitutively and secreted in an active form. The enzyme was purified to homogeneity by (NH4)2SO4 fractionation, anion exchange and affinity chromatography. The protein had a correct N-termin...


Mass-murder deletion of 19 ORFs from Saccharomyces cerevisiae chromosome XI  

Microsoft Academic Search

Nineteen open reading frames (ORFs) in the left arm of chromosome XI of the yeast Saccharomyces cerevisiae were inactivated. This was done by producing single-gene or contiguous-gene deletions in haploid and diploid strains. Four deletions are lethal to the corresponding haploid strains, and two result in a failure to grow on a rich glycerol medium. Complementation experiments showed that five

Micheline Vandenbol; Cécile Fairhead



Physiological effects of nitrogen starvation in an anaerobic batch culture of Saccharomyces cerevisiae  

Microsoft Academic Search

The effects of nitrogen starvation on the anaerobic physiology of Saccharomyces cerevisiae were studied in cells cultivated in a bioreactor. The composition of the mineral medium was designed such that the nitrogen source became depleted while there was still ample glucose left in the medium. The culture was characterized by acoustic gas analysis, flow injection analysis and HPLC analysis of

Ulrik Schulze; G. Liden; J. Nielsen; J. Villadsen



Growth of Saccharomyces cerevisiae in a chemostat under high glucose conditions  

Microsoft Academic Search

Saccharomyces cerevisiae was grown in a chemostat under high glucose conditions (up to 300 g l-1). The results support the view that higher glucose feed favors higher ethanol production regardless of the existence of osmotic stress. A low glucose utilization and yield coefficient provides an opportunity to improve continuous fermentation performance in the fuel alcohol industry. The possibility exists of reusing

Yupeng Zhao; Yen-Han Lin



Deterministic mathematical models of the cAMP pathway in Saccharomyces cerevisiae  

Microsoft Academic Search

BACKGROUND: Cyclic adenosine monophosphate (cAMP) has a key signaling role in all eukaryotic organisms. In Saccharomyces cerevisiae, it is the second messenger in the Ras\\/PKA pathway which regulates nutrient sensing, stress responses, growth, cell cycle progression, morphogenesis, and cell wall biosynthesis. A stochastic model of the pathway has been reported. RESULTS: We have created deterministic mathematical models of the PKA

Thomas Williamson; Jean-Marc Schwartz; Douglas B Kell; Lubomira Stateva



Influence of thermal and osmotic stresses on the viability of the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

This work studies the effect of thermal and dehydration kinetics on the viability of Saccharomyces cerevisiae. The influence of the rate of temperature (T) and osmotic pressure (?) increases are first investigated. Results showed that yeast viability is preserved by slow variations of temperature or osmotic pressure in a precise range of T or ?. The influence of a previous

Laurent Beney; Iñigo Mart??nez de Marañón; Pierre-André Marechal; Patrick Gervais



Heat Shock Protein Genes and Newly Integrated Glucose Metabolic Pathways Promote Ethanol Tolerance of Saccharomyces cerevisiae  

Technology Transfer Automated Retrieval System (TEKTRAN)

Lignocellulose-to-ethanol conversion provides a promising alternative means for production of sustainable and cleaner transportation fuels. Development of stress tolerant ethanologenic Saccharomyces cerevisiae is important for low-cost biobased economy. Tolerance to high levels of ethanol has been...


Chromosomal integration of recombinant alpha-amylase and glucoamylase genes in saccharomyces cerevisiae for starch conversion  

Technology Transfer Automated Retrieval System (TEKTRAN)

Recombinant constructs of barley '-amylase and Lentinula edodes glucoamylase genes were integrated into the chromosomes of Saccharomyces cerevisiae. The insertion was confirmed by PCR amplification of the gene sequence in the chromosomes. The expression was analyzed by SDS-PAGE of the enzymes puri...



Technology Transfer Automated Retrieval System (TEKTRAN)

The gene coding for Lentinus edodes glucoamylase (GLA) was isolated by PCR of the cDNA obtained by transcribing the total mRNA collected from the mycelium. The GLA gene was cloned into Saccharomyces cerevisiae, expressed and constitutively secreted in an active form. The enzyme was purified to hom...


High Osmolarity Extends Life Span in Saccharomyces cerevisiae by a Mechanism Related to Calorie Restriction  

Microsoft Academic Search

Calorie restriction (CR) extends life span in many different organisms, including mammals. We describe here a novel pathway that extends the life span of Saccharomyces cerevisiae mother cells but does not involve a reduction in caloric content of the media, i.e., there is growth of yeast cells in the presence of a high concentration of external osmolytes. Like CR, this

Matt Kaeberlein; Alex A. Andalis; Gerald R. Fink; Leonard Guarente



Saccharomyces cerevisiae in the stationary phase as a model organism — characterization at cellular and proteome level  

Microsoft Academic Search

The yeast Saccharomyces cerevisiae has been used as a model organism to investigate responses to different environmental stressors. The importance of their conclusions has been expanded to human cells. The experiments were done with exponentially growing cells, which do not resemble human cells. Human and other eukaryotic cells spend the greater part of their lives in a quiescent state, known

Teja Zakrajšek; Peter Raspor; Polona Jamnik



Ethanol-Independent Biofilm Formation by a Flor Wine Yeast Strain of Saccharomyces cerevisiae?  

PubMed Central

Flor strains of Saccharomyces cerevisiae form a biofilm on the surface of wine at the end of fermentation, when sugar is depleted and growth on ethanol becomes dependent on oxygen. Here, we report greater biofilm formation on glycerol and ethyl acetate and inconsistent formation on succinic, lactic, and acetic acids.

Zara, Severino; Gross, Michael K.; Zara, Giacomo; Budroni, Marilena; Bakalinsky, Alan T.



Saccharomyces cerevisiae JEN1 Promoter Activity Is Inversely Related to Concentration of Repressing Sugar  

Microsoft Academic Search

When carbon sources are changed, Saccharomyces cerevisiae transcriptional patterns drastically change. To identify genes whose transcription can be used to quantitatively measure sugar concentrations, we searched genomic expression databases for a set of genes that are highly induced during the diauxic shift, and we used the promoters from these genes to drive expression of green fluorescent protein (GFP). Certain sugars,

Prima Chambers; Aminatu Issaka; Sean P. Palecek



Display of wasp venom allergens on the cell surface of Saccharomyces cerevisiae  

Microsoft Academic Search

BACKGROUND: Yeast surface display is a technique, where the proteins of interest are expressed as fusions with yeast surface proteins and thus remain attached to the yeast cell wall after expression. Our purpose was to study whether allergens expressed on the cell surface of baker's yeast Saccharomyces cerevisiae preserve their native allergenic properties and whether the yeast native surface glycoproteins

Irina Borodina; Bettina M Jensen; Ib Søndergaard; Lars K Poulsen



Oral administration of Saccharomyces cerevisiae boulardii reduces Escherichia coli endotoxin associated mortality in weaned pigs  

Technology Transfer Automated Retrieval System (TEKTRAN)

The effects of active dry yeast, Saccharomyces cerevisiae boulardii (Scb), on the immune/neuroendocrine response and subsequent mortality to E. coli lipopolysaccharide (LPS) administration were evaluated in newly weaned pigs (26.1 + or - 3.4 d of age). Barrows were assigned to 1 of 2 treatment group...


Behaviour of Saccharomyces cerevisiae cells entrapped in a polyacrylamide gel and performing alcoholic fermentation  

Microsoft Academic Search

The behaviour of Saccharomyces cerevisiae cells entrapped in a polyacrylamide gel was studied during their continuous function in an ethanol-producing reactor. Polymerization destroys 40% to 80% of the cells, depending on their physiological state. A three day adaptation phase is required before ethanol production stabilizes and this phase corresponds to an increase in cell concentration in the gels and to

M. H. Siess; C. Divies



Biotechnology of natural and winery-associated strains of Saccharomyces cerevisiae  

Microsoft Academic Search

A new body of evidence challenges the original consolidated theory of Pasteur on the natural (vineyard) origin of wine strains of Saccharomyces cerevisiae and instead indicates a local, winery-restricted life cycle. The findings open novel biotechnological perspectives for obtaining autochthonous selected starters for the wine industry. A local, individual, and specific fermenting yeast flora, mass selected year after year through

Alessandro Martini



Enological and genetic traits of Saccharomyces cerevisiae isolated from former and modern wineries  

Microsoft Academic Search

Saccharomyces cerevisiae strains isolated from two different wineries in central Italy were subjected to enological and molecular characterization to investigate the influence of the winery environment. One of the selected wineries is a modern, working winery, whereas the second one was abandoned since 1914 and was located in an artificial cavern. The results obtained by our analysis of the fermentation

Luca Cocolin; Vincenzo Pepe; Francesca Comitini; Giuseppe Comi; Maurizio Ciani



Genetic diversity of wine Saccharomyces cerevisiae strains in an experimental winery from Galicia (NW Spain)  

Microsoft Academic Search

Genetic diversity of wine Saccharomyces cerevisiae strains involved in spontaneous fermentations was studied by analysis of mitochondrial DNA restriction patterns. Yeasts were isolated at different stages of fermentations with must from three different white grapevine varieties, Albariño, Godello and Treixadura, which are autochthonous from Galicia. Nineteen different patterns out of a total of 446 strains analysed were identified, but only

P. Blanco; A. Ramilo; M. Cerdeira; I. Orriols



Cycles of mitochondrial energization driven by the ultradian clock in a continuous culture of Saccharomyces cerevisiae  

Microsoft Academic Search

A continuous culture of Saccharomyces cerevisiae IFO 0233, growing with glucose as the major carbon and energy source, shows oscillations of respiration with a period of 48 min. Samples taken at maxima and minima indicate that (i) periodic changes do not occur as a result of carbon depletion, (ii) intrinsic differences in respiratory activity occur in washed organisms and (iii)

David Lloyd; L. Eshantha; J. Salgado; Michael P. Turner; Marc T. E. Suller; Douglas Murray



Role of Hexose Transport in Control of Glycolytic Flux in Saccharomyces cerevisiae  

Microsoft Academic Search

The yeast Saccharomyces cerevisiae predominantly ferments glucose to ethanol at high external glucose concentrations, irrespective of the presence of oxygen. In contrast, at low external glucose concentrations and in the presence of oxygen, as in a glucose-limited chemostat, no ethanol is produced. The importance of the external glucose concentration suggests a central role for the affinity and maximal transport rates

Karin Elbing; Christer Larsson; Roslyn M. Bill; Eva Albers; Jacky L. Snoep; Eckhard Boles; Stefan Hohmann; Lena Gustafsson



HOCl-mediated cell death and metabolic dysfunction in the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

The nature of oxidative damage to Saccharomyces cerevisiae caused by levels of HOCl that inhibit cell replication was explored with the intent of identifying the loci of lethal lesions. Functions of cytosolic enzymes and organelles that are highly sensitive to inactivation by HOCl, including aldolase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and the mitochondrion, were only marginally affected by exposure of the yeast

David A King; Diane M Hannum; Jian-Shen Qi; James K Hurst




EPA Science Inventory

We have sequenced the structural gene and flanking regions for lanosterol 14 alpha-demethylase (14DM) from Saccharomyces cerevisiae. An open reading frame of 530 codons encodes a 60.7-kDa protein. When this gene is disrupted by integrative transformation, the resulting strain req...



EPA Science Inventory

We have sequenced the structural gene and flanking regions for lanosterol 14oc-demethylase (14DM) from Saccharomyces cerevisiae. n open reading fram of 530 codons encodes a 60.7-kDa protein. hen this gene is disrupted by integrative transformation, the resulting strain requires e...


Evidence for Sterilization of Saccharomyces Cerevisiae K 7 by an External Magnetic Flux  

Microsoft Academic Search

A new process of sterilization on Saccharomyces cerevisiae proposed and experimentally demonstrated. This process consists of external magnetic flux and ferrite. Consequently, an alteration of yeast cells, caused by treatment with 2000 G magnetic flux and 6 g of ferrite, was detected through observation of the release of materials from yeast cells absorbing at 260 nm and microscopy of disrupted

Tatsuya Ito; Yuzo Murayama; Masafumi Suzuki; Noboru Yoshimura; Kimio Iwano; Kozo Kudo



The effect of supplementing cows with live yeast, Saccharomyces cerevisiae, on ciliate fauna and ruminal fermentation  

Microsoft Academic Search

The infl uence of a live culture of Saccharomyces cerevisiae on the number of ciliates, activity of carboxymethylcellulose- (CMC), xylan- and starch-degrading enzymes, as well as the concentration of VFA in the rumen of two cows was examined. The animals were fed either hay and concentrate alone or supplemented with live yeast given at the rate of 5 g\\/d. It

B. Kowalik; T. Micha?owski; J. J. Paj?k; M. Taciak; J. Rawa


Effects of yeast culture ( Saccharomyces cerevisiae) on ruminal digestion in non-lactating dairy cows  

Microsoft Academic Search

Six non-lactating dairy cows fitted with ruminal cannulas were used in a cross-over design, to investigate the effects of supplemental yeast culture (Saccharomyces cerevisiae) (YC) and interaction of YC by sampling time on ruminal fermentation and in situ fibre degradation. Cows were fed twice daily with a diet composed of 67% corn silage, 32% concentrate and 1% vitamin and mineral

F. Enjalbert; J. E. Garrett; R. Moncoulon; C. Bayourthe; P. Chicoteau



Genome-wide identification of Saccharomyces cerevisiae genes required for tolerance to acetic acid  

Microsoft Academic Search

BACKGROUND: Acetic acid is a byproduct of Saccharomyces cerevisiae alcoholic fermentation. Together with high concentrations of ethanol and other toxic metabolites, acetic acid may contribute to fermentation arrest and reduced ethanol productivity. This weak acid is also a present in lignocellulosic hydrolysates, a highly interesting non-feedstock substrate in industrial biotechnology. Therefore, the better understanding of the molecular mechanisms underlying S.

Nuno P Mira; Margarida Palma; Joana F Guerreiro; Isabel Sá-Correia



Yeh1 Constitutes the Major Steryl Ester Hydrolase under Heme-Deficient Conditions in Saccharomyces cerevisiae  

Microsoft Academic Search

Steryl esters are stored in intracellular lipid droplets from which they are mobilized upon demand and hydrolyzed to yield free sterols and fatty acids. The mechanisms that control steryl ester mobilization are not well understood. We have previously identified a family of three lipases of Saccharomyces cerevisiae that are required for efficient steryl ester hydrolysis, Yeh1, Yeh2, and Tgl1 (R.

R. Koffel; R. Schneiter




Technology Transfer Automated Retrieval System (TEKTRAN)

This work demonstrates the first example of a fungal LDH expressed in yeast. A L(+)-lactate dehydrogenase gene, ldhA, from the filamentous fungus Rhizopus oryzae was modified to be expressed under control of the Saccharomyces cerevisiae adhl promoter and terminator, then placed in a 2 micron contai...


A mathematical model of the mating signal transduction pathway in the yeast Saccharomyces cerevisiae. Final report  

SciTech Connect

Outline of two major goals in my proposal for this fellowship. First goal having no previous training in biology, was to become knowledgeable of the paradigms, experimental techniques, and current research interests of molecular biology. Second goal was to construct a mathematical model of the mating signal transduction pathway in the yeast Saccharomyces cerevisiae.

Thomas Ivan Milac



Proof and evolutionary analysis of ancient genome duplication in the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

Whole-genome duplication followed by massive gene loss and specialization has long been postulated as a powerful mechanism of evolutionary innovation. Recently, it has become possible to test this notion by searching complete genome sequence for signs of ancient duplication. Here, we show that the yeast Saccharomyces cerevisiae arose from ancient whole-genome duplication, by sequencing and analysing Kluyveromyces waltii, a related

Bruce W. Birren; Manolis Kellis; Eric S. Lander



Polyene Resistance and the Isolation of Sterol Mutants in Saccharomyces cerevisiae  

Microsoft Academic Search

SUMMARY Mutants of Saccharomyces cerevisiae have been selected for resistance to the polyene antibiotics, etruscomycin, filipin, nystatin, pimaricin and rimocidin. All mutants are resistant to nystatin and there are several patterns of cross-resistance. The mutants were allocated to four unlinked genes pol I, 2, 3 and 5. A fifth gene, pol 4, was recovered as a double mutant with a




Engineering Saccharomyces cerevisiae to produce feruloyl esterase for the release of ferulic acid from switchgrass  

Technology Transfer Automated Retrieval System (TEKTRAN)

The Aspergillus niger ferulic acid esterase gene (faeA) was cloned into Saccharomyces cerevisiae via a yeast expression vector, resulting in efficient expression and secretion of the enzyme in the medium. The recombinant enzyme was purified to homogeneity by anion-exchange and hydrophobic interactio...


Role of Heat Shock Transcription Factor in Saccharomyces cerevisiae Oxidative Stress Response  

Microsoft Academic Search

The heat shock transcription factor Hsf1 of the yeast Saccharomyces cerevisiae regulates the transcription of a set of genes that contain heat shock elements (HSEs) in their promoters and function in diverse cellular processes, including protein folding. Here, we show that Hsf1 activates the transcription of various target genes when cells are treated with oxidizing reagents, including the superoxide anion

Ayako Yamamoto; Junko Ueda; Noritaka Yamamoto; Naoya Hashikawa; Hiroshi Sakurai



Genomewide Screening for Genes Associated with Gliotoxin Resistance and Sensitivity in Saccharomyces cerevisiae  

Microsoft Academic Search

Gliotoxin (GT) is a secondary fungal metabolite with pleiotropic immunosuppressive properties that have been implicated in Aspergillus virulence. However, the mechanisms of GT cytotoxicity and its molecular targets in eukaryotic cells have not been fully characterized. We screened a haploid library of Saccharomyces cerevisiae single-gene deletion mutants (4,787 strains in EUROSCARF) to identify nonessential genes associated with GT increased resistance

Georgios Chamilos; Russell E. Lewis; Gregory A. Lamaris; Nathaniel D. Albert; Dimitrios P. Kontoyiannis



Theme and Variation Among Silencing Proteins in Saccharomyces cerevisiae and Kluyveromyces lactis  

Microsoft Academic Search

The cryptic mating type loci in Saccharomyces cerevisiae act as reservoirs of mating type information used in mating type switching in homothallic yeast strains. The transcriptional silencing of these loci depends on the formation of a repressive chromatin structure that is reminiscent of heterochromatin. Silent information regulator (Sir) proteins 2-4 are absolutely required for silencing. To learn more about silencing,

Stefan U. Astrom; Jasper Rine


Cane molasses fermentation for continuous ethanol production in an immobilized cells reactor by Saccharomyces cerevisiae  

Microsoft Academic Search

Sodium-alginate immobilized yeast was employed to produce ethanol continuously using cane molasses as a carbon source in an immobilized cell reactor (ICR). The immobilization of Saccharomyces cerevisiae was performed by entrapment of the cell cultured media harvested at exponential growth phase (16 h) with 3% sodium alginate. During the initial stage of operation, the ICR was loaded with fresh beads of

Farshid Ghorbani; Habibollah Younesi; Abbas Esmaeili Sari; Ghasem Najafpour



Ethanol fermentation in a magnetically fluidized bed reactor with immobilized Saccharomyces cerevisiae in magnetic particles  

Microsoft Academic Search

Ethanol fermentation by immobilized Saccharomyces cerevisiae cells in magnetic particles was successfully carried out in a magnetically stabilized fluidized bed reactor (MSFBR). These immobilized magnetic particles solidified in a 2 % CaCl2 solution were stable and had high ethanol fermentation activity. The performance of ethanol fermentation of glucose in the MSFBR was affected by initial particle loading rate, feed sugar

Chun-Zhao Liu; Feng Wang; Fan Ou-Yang



Ethanol fermentation of a diluted molasses medium by Saccharomyces cerevisiae immobilized on chrysotile  

Microsoft Academic Search

In this work, the catalytic role of chrysotile support on the acceleration of alcoholic fermentation under non-aseptic conditions by Saccharomyces cerevisiae was investigated. The fermentation medium employed consisted only of diluted sugar-cane molasses. In the batch fermentations process with immobilized yeasts, the initial rate of CO 2 production increased roughly 27 % during the first 30 minutes, compared to systems

Ranulfo Monte Alegre; Maurício Rigo; Inés Joekes



Transcriptome profiling of Saccharomyces cerevisiae mutants lacking C2H2 zinc finger proteins  

Microsoft Academic Search

BACKGROUND: The budding yeast Saccharomyces cerevisiae is a eukaryotic organism with extensive genetic redundancy. Large-scale gene deletion analysis has shown that over 80% of the ~6200 predicted genes are nonessential and that the functions of 30% of all ORFs remain unclassified, implying that yeast cells can tolerate deletion of a substantial number of individual genes. For example, a class of

Jinghe Mao; Tanwir Habib; Ming Shenwu; Baobin Kang; Wilbur Allen; LaShonda Robertson; Jack Y Yang; Youping Deng



Effects of glucose and mannose on the flocculation behavior of Saccharomyces cerevisiae at different life stages  

Microsoft Academic Search

The temporal flocculation behavior of Saccharomyces cerevisiae at different life stages is investigated using glucose and mannose as the different carbon sources, and the temporal variations of cell size, zeta potential and stability ratio of cell suspension are measured. It is found that the largest cell size and the lowest stability ratio of cell suspension occurred at the middle period

You-Im Chang; Lian-Hua Shih; Shyh-Wei Chen



Functional genomic analysis of a commercial wine strain of Saccharomyces cerevisiae under differing nitrogen conditions  

Microsoft Academic Search

DNA microarray analysis was used to profile gene expression in a commercial isolate of Saccharomyces cerevisiae grown in a synthetic grape juice medium under conditions mimicking a natural environment for yeast: High-sugar and variable nitrogen conditions. The high nitrogen condition displayed elevated levels of expression of genes involved in biosynthesis of macromolecular precursors across the time course as compared to

Leilah E. Backhus; Joseph DeRisi; Patrick O. Brown; Linda F. Bisson



Conservation of glycolytic oscillations in Saccharomyces cerevisiae and human pancreatic ? ? ? ? ?-cells: a study of metabolic robustness  

Microsoft Academic Search

The present study compares two computer models of the first part of glucose catabolism in different organisms in search of evolutionarily conserved characteristics of the glycolysis cycle and pro- poses the main parameters that define the stable steady-state or oscilla- tory behavior of the glycolytic system. It is suggested that in both human pancreatic ?-cells and Saccharomyces cerevisiae there are

A. S. Silva; J. A. Yunes



Internal Trehalose Protects Endocytosis from Inhibition by Ethanol in Saccharomyces cerevisiae  

Microsoft Academic Search

Endocytosis in Saccharomyces cerevisiae is inhibited by concentrations of ethanol of 2 to 6% (vol\\/vol), which are lower than concentrations commonly present in its natural habitats. In spite of this inhibition, endocytosis takes place under enological conditions when high concentrations of ethanol are present. Therefore, it seems that yeast has developed some means to circumvent the inhibition. In this work

P. Lucero; E. Penalver; E. Moreno; R. Lagunas



Flux analysis of recombinant Saccharomyces cerevisiae YPB-G utilizing starch for optimal ethanol production  

Microsoft Academic Search

Genetically modified Saccharomyces cerevisiae strain (YPB-G) which secretes a bifunctional fusion protein that contains both Bacillus subtilis ?-amylase and Aspergillus awamori glucoamylase activities was used for the direct conversion of starch into ethanol. Starch was either supplied initially to different nutrient media or added instantaneously to the reactor at various discrete time instants (pulse feeding). Stoichiometric modeling was used to

Tunahan Çak?r; K. Yalç?n Arga; M. Mete Alt?nta?; Kutlu Ö. Ülgen



In vivo activation by ethanol of plasma membrane ATPase of Saccharomyces cerevisiae  

Microsoft Academic Search

Ethanol, in concentrations that affect growth and fermentation rates (3 to 10% (vol\\/vol)), activated in vivo the plasma membrane ATPase of Saccharomyces cerevisiae. The maximal value for this activated enzyme in cells grown with 6 to 8% (vol\\/vol) ethanol was three times higher than the basal level (in cells grown in the absence of ethanol). The Km values for ATP,

M. F. Rosa; I. Sa-Correia



Genes involved in the control of nuclear fusion during the sexual cycle of Saccharomyces cerevisiae  

Microsoft Academic Search

Mutants of Saccharomyces cerevisiae defective for nuclear fusion have been isolated. Their mutations have been characterized by meiotic analysis, dominance-recessivity and complementation. Twelve of the mutations are allelic to the previously described kar 1-1; five affect a second gene designated KAR 2 and three affect a third gene designated KAR 3. There is evidence suggesting that other two mutants are

Julio Polaina; Jaime Conde



Autonomy of Mitochondria in Saccharomyces cerevisiae in their Production of Messenger RNA  

Microsoft Academic Search

In ts-136, a temperature-sensitive mutant of Saccharomyces cerevisiae, nuclear and mitochondrial RNA production can be inhibited selectively by exposure to 36 degrees and ethidium bromide, respectively. Using the programming of mitochondrial polysomes, as measured by their ability to form nascent polypeptide chains, as an assay for functional messenger RNA, we have determined its response to temperature shifts and ethidium bromide.

Henry R. Mahler; Karl Dawidowicz



Cytoplasmic petite induction in recombination-deficient mutants of Saccharomyces cerevisiae  

Microsoft Academic Search

As compared to the original wild type, the induction of the cytoplasmic ; petite'' mutation by ultraviolet light and by the intercalating dye, ethidium ; bromide, is reduced in two mutants (rec4 and rec5) of Saccharomyces cerevisiae. ; These mutants are blocked in x-ray or ultraviolet light-induced intragenic ; recondbination. It then appears that the products of nuclear genes necessary




Trehalose reserve in Saccharomyces cerevisiae: phenomenon of transport, accumulation and role in cell viability  

Microsoft Academic Search

Strains of Saccharomyces cerevisiae deleted for TPS1 encoding trehalose-6-phosphate synthase still accumulate trehalose when harbouring a functional MAL locus. We demonstrate that this accumulation results from an active uptake of trehalose present in the ‘yeast extract’ used to make the enriched culture media and that no accumulation is observed in mineral media. The uptake of trehalose was shown to be

L Plourde-Owobi; S Durner; G Goma; J François



Dependence of the maximum temperature for growth of Saccharomyces cerevisiae on nutrient concentration  

Microsoft Academic Search

Saccharomyces cerevisiae was grown in a chemostat under glucose limitation at three superoptimal temperatures. In each steady state the specific growth rate was the sum of the dilution rate and the specific death rate, exponential death concurring with exponential growth. The specific death rate was a function of the temperature while the specific growth rate was a function of both

N. VAN UDEN; A. Madeira-Lopes



Screening of a soil metatranscriptomic library by functional complementation of Saccharomyces cerevisiae mutants  

Microsoft Academic Search

Metatranscriptomics applied to environmental transcripts provides unique opportunities to reveal microbial activity in the environment and to discover novel enzymes of potential use in biotechnological applications. Here, by functional complementation of a pho5? mutation (affecting a repressible acid phosphatase) and a his3? mutation in Saccharomyces cerevisiae, we identified fungal genes encoding an acid phosphatase and an imidazoleglycerol-phosphate dehydratase in a

Harald Kellner; Patricia Luis; Daniel Portetelle; Micheline Vandenbol



Mitochondrial Genomic Dysfunction Causes Dephosphorylation of Sch9 in the Yeast Saccharomyces cerevisiae?†  

PubMed Central

TORC1-dependent phosphorylation of Saccharomyces cerevisiae Sch9 was dramatically reduced upon exposure to a protonophore or in respiration-incompetent ?0 cells but not in respiration-incompetent pet mutants, providing important insight into the molecular mechanisms governing interorganellar signaling in general and retrograde signaling in particular.

Kawai, Shigeyuki; Urban, Jorg; Piccolis, Manuele; Panchaud, Nicolas; De Virgilio, Claudio; Loewith, Robbie



Genetics of X-Ray Induced Double Strand Break Repair in Saccharomyces Cerevisiae.  

National Technical Information Service (NTIS)

The possible fates of x-ray-induced double-strand breaks in Saccharomyces cerevisiae were examined. One possible pathway which breaks can follow, the repair pathway, was studied by assaying strains with mutations in the RAD51, RAD54, and RAD57 loci for do...

M. E. Budd



Chromosome healing through terminal deletions generated by de novo telomere additions in Saccharomyces cerevisiae  

Microsoft Academic Search

Broken chromosomes healed by de novo addition of a telomere are a major class of genome rearrangements seen in Saccharomyces cerevisiae and similar to rearrangements seen in human tumors. We have analyzed the sequences of 534 independent de novo telomere additions within a 12-kb region of chromosome V. The distribution of events mirrored that of four-base sequences consisting of the

Christopher D. Putnam; Vincent Pennaneach; Richard D. Kolodner



End3p-Mediated Endocytosis Is Required for Spore Wall Formation in Saccharomyces cerevisiae  

Microsoft Academic Search

During sporulation in Saccharomyces cerevisiae, vesicles transported to the vicinity of spindle pole bodies are fused to each other to generate bilayered prospore membranes (PSMs). PSMs encapsulate the haploid nuclei that arise from the meiotic divisions and serve as platforms for spore wall deposition. Membrane trafficking plays an important role in supplying vesicles for these processes. The endocytosis-deficient mutant, end3,

Masayo Morishita; JoAnne Engebrecht



A Saccharomyces cerevisiae Genome-Wide Mutant Screen for Altered Sensitivity to K1 Killer Toxin  

Microsoft Academic Search

Using the set of Saccharomyces cerevisiae mutants individually deleted for 5718 yeast genes, we screened for altered sensitivity to the antifungal protein, K1 killer toxin, that binds to a cell wall -glucan receptor and subsequently forms lethal pores in the plasma membrane. Mutations in 268 genes, including 42 in genes of unknown function, had a phenotype, often mild, with 186

Nicolas Page ´; Manon Gerard-Vincent; Patrice Menard; Maude Beaulieu; Masayuki Azuma; Gerrit J. P. Dijkgraaf; Thuy Nguyen; Tim Dowse; Anne-Marie Sdicu; Howard Bussey


Unwinding RNA in Saccharomyces cerevisiae: DEAD-box proteins and related families  

Microsoft Academic Search

Members of the RNA-helicase family are defined by several evolutionary conserved motifs. They are found in all organisms – from bacteria to humans – and many viruses. The minimum number of RNA helicases present within a eukaryotic cell can be predicted from the complete sequence of the Saccharomyces cerevisiae genome. Recent progress in the functional analysis of various family members

Jesús de la Cruz; Dieter Kressler; Patrick Linder



The Effect of Sodium Azide on Basic and Induced Thermotolerance in Saccharomyces cerevisiae  

Microsoft Academic Search

The action mechanism of the mitochondrial inhibitor sodium azide on thermotolerance in Saccharomyces cerevisiae was studied. At ambient growth temperature, pretreatment with sodium azide was shown to improve the thermotolerance of parent cells and the hsp104 mutant. Treating with the inhibitor during a mild heat shock suppressed the development of induced thermotolerance due to the inhibition of heat shock protein

E. I. Rachenko; E. G. Rikhvanov; N. N. Varakina; T. M. Rusaleva; G. B. Borovskii; V. K. Voinikov



Stability of the plasma membrane in Saccharomyces cerevisiae enriched with phosphatidylcholine or phosphatidylethanolamine.  

PubMed Central

Spheroplasts from Saccharomyces cerevisiae NCYC 366, enriched in phosphatidylethanolamine after growth in medium supplemented with 1 mM ethanolamine, were more resistant to osmotic lysis than were spheroplasts from cells grown in the presence of 1 mM choline and enriched in phosphatidylcholine.

Hossack, J A; Sharpe, V J; Rose, A H



Tryptophan Permease Gene TAT2 Confers High-Pressure Growth in Saccharomyces cerevisiae  

Microsoft Academic Search

Hydrostatic pressure in the range of 15 to 25 MPa was found to cause arrest of the cell cycle in G1 phase in an exponentially growing culture of Saccharomyces cerevisiae, whereas a pressure of 50 MPa did not. We found that a plasmid carrying the TAT2 gene, which encodes a high-affinity tryptophan permease, enabled the cells to grow under conditions




Low frequency thermo-ultrasonication of Saccharomyces cerevisiae suspensions: Effect of temperature and of ultrasonic power  

Microsoft Academic Search

The combined effect of low frequency ultrasound (20 kHz) with temperature on the survival of a strain of Saccharomyces cerevisiae suspended in water was studied. The treatment temperatures tested were 45 °C, 50 °C and 55 °C; the actual ultrasonic powers tested were 50 W, 100 W and 180 W. Application of ultrasonic waves at a non-lethal temperature did not

L. Ciccolini; P. Taillandier; A. M. Wilhem; H. Delmas; P. Strehaiano




Technology Transfer Automated Retrieval System (TEKTRAN)

Fusarium solani f. sp. glycines (Fsg) is in the same fungal class as Saccharomyces cerevisiae. To compare the genes of these two fungi, cDNA was synthesized by reverse transcription from total RNA extracted from a culture of Fsg. cRNA was synthesized from cDNA and labeled with biotin by in vitro tra...


Metabolism and selected functions of sphingolipids in the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

Our knowledge of sphingolipid metabolism and function in Saccharomyces cerevisiae is growing rapidly. Here we discuss the current status of sphingolipid metabolism including recent evidence suggesting that exogenous sphingoid long-chain bases must first be phosphorylated and then dephosphorylated before incorporation into ceramide. Phenotypes of strains defective in sphingolipid metabolism are discussed because they provide hints about the undiscovered functions of

Robert C Dickson; Robert L Lester



A mutation affecting lipoamide dehydrogenase, pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase activities in Saccharomyces cerevisiae  

Microsoft Academic Search

In Saccharomyces cerevisiae a nuclear recessive mutation, lpd1, which simultaneously abolishes the activities of lipoamide dehydrogenase, 2-oxoglutarate dehydrogenase and pyruvate dehydrogenase has been identified. Strains carrying this mutation can grow on glucose or poorly on ethanol, but are unable to grow on media with glycerol or acetate as carbon source. The mutation does not prevent the formation of other tricarboxylic

J. Richard Dickinson; Douglas J. Roy; Ian W. Dawes



Regulation of Nuclear Division and Ascosporogenesis in Apomictic Strains of Saccharomyces cerevisiae  

Microsoft Academic Search

Effects of nutritional alterations (carbon source, zinc) on nuclear division and protein synthesis during apomictic and meiotic differentiation in Saccharomyces cerevisiae strain 19el were investigated. The approach taken has led to identification of: physiological prerequisites for both developmental routes, landmark cytological and molecular events controlling the manner of nuclear division and spore production, other environmental modifications suppressing apomixis, and the

Carl Anthony Bilinski



Ssd1 Is Required for Thermotolerance and Hsp104-Mediated Protein Disaggregation in Saccharomyces cerevisiae  

Microsoft Academic Search

In the budding yeast Saccharomyces cerevisiae, the Hsp104-mediated disaggregation of protein aggregates is essential for thermotolerance and to facilitate the maintenance of prions. In humans, protein aggregation is associated with neuronal death and dysfunction in many neurodegenerative diseases. Mechanisms of aggre- gation surveillance that regulate protein disaggregation are likely to play a major role in cell survival after acute stress.

Snober S. Mir; David Fiedler; Anil G. Cashikar



Genome-wide expression profile of steroid response in Saccharomyces cerevisiae  

Microsoft Academic Search

The response of the yeast Saccharomyces cerevisiae to human steroid hormone progesterone was studied by genomic expression profiling. The transcription profile data revealed that steroid response was a global phenomenon wherein a host of genes were affected. For example, 163 genes were upregulated and 40 genes were downregulated, by at least more than twofold. The major categories of upregulated genes

Dibyendu Banerjee; Beena Pillai; Neerja Karnani; Gauranga Mukhopadhyay; Rajendra Prasad



Nuclease Activity of Saccharomyces cerevisiae Mre11 Functions in Targeted Nucleotide Alteration  

Microsoft Academic Search

Oligonucleotides can be used to direct site-specific changes in genomic DNA through a process in which mismatched base pairs in the oligonucleotide and the target DNA are created. The mechanism by which these complexes are developed and resolved is being studied by using Saccharomyces cerevisiae as a model system. Genetic analyses have revealed that in all likelihood the reaction occurs

Li Liu; Michael Usher; Yiling Hu; Eric B. Kmiec



Functional Assembly of Minicellulosomes on the Saccharomyces cerevisiae Cell Surface for Cellulose Hydrolysis and Ethanol Production  

Microsoft Academic Search

We demonstrated the functional display of a miniscaffoldin on the Saccharomyces cerevisiae cell surface consisting of three divergent cohesin domains from Clostridium thermocellum (t), Clostridium cellulolyticum (c), and Ruminococcus flavefaciens (f). Incubation with Escherichia coli lysates containing an endoglucanase (CelA) fused with a dockerin domain from C. thermocellum (At), an exoglucanase (CelE) from C. cellulolyticum fused with a dockerin domain

Shen-Long Tsai; Jeongseok Oh; Shailendra Singh; Ruizhen Chen; Wilfred Chen



Structural Studies of the Final Enzyme in the ?-Aminoadipate Pathway-Saccharopine Dehydrogenase from Saccharomyces cerevisiae  

Microsoft Academic Search

The 1.64 Å structure of the apoenzyme form of saccharopine dehydrogenase (SDH) from Saccharomyces cerevisiae shows the enzyme to be composed of two domains with similar dinucleotide binding folds with a deep cleft at the interface. The structure reveals homology to alanine dehydrogense, despite low primary sequence similarity. A model of the ternary complex of SDH, NAD, and saccharopine identifies residues

D. L. Burk; J. Hwang; E. Kwok; L. Marrone; V. Goodfellow; G. I. Dmitrienko; A. M. Berghuis



Trans-acting regulatory variation in Saccharomyces cerevisiae and the role of transcription factors  

Microsoft Academic Search

Natural genetic variation can cause significant differences in gene expression, but little is known about the polymorphisms that affect gene regulation. We analyzed regulatory variation in a cross between laboratory and wild strains of Saccharomyces cerevisiae. Clustering and linkage analysis defined groups of coregulated genes and the loci involved in their regulation. Most expression differences mapped to trans-acting loci. Positional

Gaël Yvert; Rachel B Brem; Jacqueline Whittle; Joshua M Akey; Eric Foss; Erin N Smith; Rachel Mackelprang; Leonid Kruglyak



Specificity of Mismatch Repair Following Transformation of Saccharomyces cerevisiae with Heteroduplex Plasmid DNA  

Microsoft Academic Search

A method is described for genetic detection of mismatch repair products following transformation of Saccharomyces cerevisiae. The method is based on the detection of beta -galactosidase activity in clonal derivatives of cells transformed with heteroduplex plasmid DNA. Heteroduplex plasmid substrates were constructed by insertion of an oligonucleotide heteroduplex into the coding sequence of the Escherichia coli lacZ gene. The plasmid

Douglas K. Bishop; Janet Andersen; Richard D. Kolodner




Microsoft Academic Search

SUMMARY Use of antibiotics as an additive in poultry diets to improve growth has been discussed in relation to bacterial resistance and the development of new products and management practices. This study was carried out to test the efficacy of a new substance (Saccharomyces cerevisiae cell walls, var. Calsberg- SCCW) obtained from the brewery industry, added (at 0.1 and 0.2%)



Reconstruction and logical modeling of glucose repression signaling pathways in Saccharomyces cerevisiae  

Microsoft Academic Search

BACKGROUND: In the yeast Saccharomyces cerevisiae, the presence of high levels of glucose leads to an array of down-regulatory effects known as glucose repression. This process is complex due to the presence of feedback loops and crosstalk between different pathways, complicating the use of intuitive approaches to analyze the system. RESULTS: We established a logical model of yeast glucose repression,

Tobias S Christensen; Ana Paula Oliveira; Jens Nielsen



A new nomenclature for the cytoplasmic ribosomal proteins of Saccharomyces cerevisiae.  

PubMed Central

The availability of the complete sequence of the Saccharomyces cerevisiae genome has allowed a comprehensive analysis of the genes encoding cytoplasmic ribosomal proteins in this organism. On the basis of this complete inventory a new nomenclature for the yeast ribosomal proteins is presented.

Mager, W H; Planta, R J; Ballesta, J G; Lee, J C; Mizuta, K; Suzuki, K; Warner, J R; Woolford, J



Arabinose and xylose fermentation by recombinant Saccharomyces cerevisiae expressing a fungal pentose utilization pathway  

Microsoft Academic Search

BACKGROUND: Sustainable and economically viable manufacturing of bioethanol from lignocellulose raw material is dependent on the availability of a robust ethanol producing microorganism, able to ferment all sugars present in the feedstock, including the pentose sugars L-arabinose and D-xylose. Saccharomyces cerevisiae is a robust ethanol producer, but needs to be engineered to achieve pentose sugar fermentation. RESULTS: A new recombinant

Maurizio Bettiga; Oskar Bengtsson; Bärbel Hahn-Hägerdal; Marie F Gorwa-Grauslund



Expression of Functional Pentameric Heat-Labile Enterotoxin B Subunit of Escherichia coli in Saccharomyces cerevisiae  

Microsoft Academic Search

Although the Escherichia coli heat-labile enterotoxin B subunit (LTB) has already been expressed in several different systems, including prokaryotic and eukaryotic organisms, studies regarding the synthesis of LTB into oligomeric structures of pentameric size in the budding yeast Saccharomyces cerevisiae have been limited. Therefore, this study used a functional signal peptide of the amylase 1A protein from rice to direct

Jung-Gu Lim; Jung-Ae Kim; Hea-Jong Chung; Tae-Geum Kim; Jung-Mi Kim; Kyung-Ryul Lee; Seung-Moon Park; Moon-Sik Yang; Dae-Hyuk Kim



Saccharomyces cerevisiae Srs2 DNA Helicase Selectively Blocks Expansions of Trinucleotide Repeats  

Microsoft Academic Search

Trinucleotide repeats (TNRs) undergo frequent mutations in families afflicted with certain neurodegenera- tive disorders and in model organisms. TNR instability is modulated both by the repeat tract itself and by cellular proteins. Here we identified the Saccharomyces cerevisiae DNA helicase Srs2 as a potent and selective inhibitor of expansions. srs2 mutants had up to 40-fold increased expansion rates of CTG,

Saumitri Bhattacharyya; Robert S. Lahue



Rapid Identification and Enumeration of Saccharomyces cerevisiae Cells in Wine by Real-Time PCR  

Microsoft Academic Search

Despite the beneficial role of Saccharomyces cerevisiae in the food industry for food and beverage production, it is able to cause spoilage in wines. We have developed a real-time PCR method to directly detect and quantify this yeast species in wine samples to provide winemakers with a rapid and sensitive method to detect and prevent wine spoilage. Specific primers were

P. Martorell; A. Querol; M. T. Fernandez-Espinar



The Ehrlich Pathway for Fusel Alcohol Production: a Century of Research on Saccharomyces cerevisiae Metabolism  

Microsoft Academic Search

Saccharomyces cerevisiae has been used for at least eight millennia in the production of alcoholic beverages (41). Along with ethanol and carbon dioxide, fermenting cultures of this yeast produce many low-molecular-weight flavor compounds. These alcohols, aldehydes, organic acids, esters, organic sul- fides, and carbonyl compounds have a strong impact on prod- uct quality. Indeed, the subtle aroma balance of these

Lucie A. Hazelwood; Jean-Marc Daran; Antonius J. A. van Maris; Jack T. Pronk; J. Richard Dickinson



Old Yellow Enzymes Protect against Acrolein Toxicity in the Yeast Saccharomyces cerevisiae  

Microsoft Academic Search

Acrolein is a ubiquitous reactive aldehyde which is formed as a product of lipid peroxidation in biological systems. In this present study, we screened the complete set of viable deletion strains in Saccharomyces cerevisiae for sensitivity to acrolein to identify cell functions involved in resistance to reactive aldehydes. We identified 128 mutants whose gene products are localized throughout the cell.

Eleanor W. Trotter; Emma J. Collinson; Ian W. Dawes; Chris M. Grant



Continuous ethanol fermentation of lactose by a recombinant flocculating Saccharomyces cerevisiae strain  

Microsoft Academic Search

Alcohol fermentation of lactose was investi- gated using a recombinant flocculating Saccharomyces cerevisiae, expressing the LAC4 (coding for b-galactosi- dase) and LAC12 (coding for lactose permease) genes of Kluyveromyces marxianus. Data on yeast fermentation and growth on a medium containing lactose as the sole carbon source are presented. In the range of studied lac- tose concentrations, total lactose consumption was

Lucília Domingues; Maria M. Dantas; Nelson Lima; José A. Teixeira



Contribution by Saccharomyces cerevisiae yeast to fermentative flavour compounds in wines from cv. Albariño  

Microsoft Academic Search

A comparative study was made of the fermentation products of Spanish Albariño wines produced with spontaneous yeast flora and an indigenous selected Saccharomyces cerevisiae strain (Alb16). The content of fermentative volatile compounds was determined by gas-chromatography-FID. Fifteen compounds (5 alcohols, 7 esters and 3 acetates) were identified in the two Albariño wines studied. Higher alcohols, ethyl esters (except ethyl hexanoate

Mar Vilanova; Carmen Sieiro



Saccharomyces cerevisiae infections and antifungal susceptibility studies by colorimetric and broth macrodilution methods  

Microsoft Academic Search

Saccharomyces cerevisiae was isolated in large numbers from operative specimens from two patients with perforated bowel and peritonitis and from the blood of another patient treated with extracorporeal membrane oxygenation. Susceptibility studies were performed on these three isolates and another 29 isolates that colonized or caused infection in a total of 19 patients seen over the last decade. All isolates

Robert N. Tiballi; Joan E. Spiegel; Lidija T. Zarins; Carol A. Kauffman



Multifactorial analysis of acetaldehyde kinetics during alcoholic fermentation by Saccharomyces cerevisiae  

Microsoft Academic Search

Acetaldehyde is the terminal electron acceptor in the alcoholic fermentation by Saccharomyces cerevisiae. Quantitatively the most important carbonyl by-product, it has relevance for ethanol production yields as well as product stabilization and toxicology. The aim of this study was to investigate the effect of various enological parameters on acetaldehyde kinetics during alcoholic fermentations. Two commercial yeast strains were tested in

J. N. Jackowetz; S. Dierschke; R. Mira de Orduña



Cross-presentation of HLA class I epitopes from influenza matrix protein produced in Saccharomyces cerevisiae  

Microsoft Academic Search

Here we report that genetically engineered yeast of the strain Saccharomyces cerevisiae expressing full-length influenza matrix protein (IMP) attached to the yeast cell wall are a very versatile host for antigen delivery. Feeding of dendritic cells with either intact yeast expressing IMP protein or soluble IMP protein cleaved off the cell wall resulted in protein uptake, processing and cross-presentation of

Andreas Wadle; Gerhard Held; Frank Neumann; Sascha Kleber; Beate Wuellner; Anne Marie Asemissen; Boris Kubuschok; Carmen Scheibenbogen; Tanja Breinig; Andreas Meyerhans; Christoph Renner



Chemical and enzymatic extraction of heavy metal binding polymers from isolated cell walls of Saccharomyces cerevisiae  

Microsoft Academic Search

Isolated cell walls of the yeast Saccharomyces cerevisiae were treated by either chemical (alkali and acid) or enzymatic (protease, mannanase or [beta]-glucuronidase) processes to yield partially purified products. These products were partially characterized by infrared analysis. They were subsequently reacted with heavy metal cation solutions and the quantity of metal accumulated by the cell wall material determined. The Cu[sup 2+

D. Brady; A. D. Stoll; L. Starke; J. R. Duncan



The use of plasmid DNA to probe DNA repair functions in the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

The survival of plasmid YRp12 treated in vitro with ultraviolet- or ?-radiation, or with restriction endonucleases, has been used to investigate in vivo RAD gene activity in Saccharomyces cerevisiae. Yields of pyrmidine dimers or single and double strand breaks in plasmid DNA were assayed by physical methods. The biological effects of these damages were assayed by transformation of wild-type cells

Charles I. White; Steven G. Sedgwick



Genetic control of plasmid DNA double-strand gap repair in yeast, Saccharomyces cerevisiae  

Microsoft Academic Search

The repair of double-strand gaps (DSGs) in the plasmid DNA of radiosensitive mutants of Saccharomyces cerevisiae has been analyzed. The proportion of repair events that resulted in complete plasmid DNA DSG recovery was close to 100% in Rad+ cells. Mutation rad55 does not influence the efficiency and preciseness of DSG repair. The mutant rad57, which is capable of recombinational DNA

V. M. Glaser; A. V. Glasunov; G. G. Tevzadze; J. R. Perera; S. V. Shestakov



Repair of double-strand breaks in plasmid DNA in the yeast Saccharomyces cerevisiae  

Microsoft Academic Search

We studied the repair of double-strand breaks (DSB) in plasmid DNA introduced into haploid cells of the yeast Saccharomyces cerevisiae. The efficiency of repair was estimated from the frequency of transformation of the cells by an autonomously replicated linearized plasmid. The frequency of “lithium” transformation of Rad+ cells was increased greatly (by 1 order of magnitude and more) compared with

Joseph R. Perera; Alexander V. Glasunov; Vadim M. Glaser; Alla V. Boreiko



Genomic Exploration of the Hemiascomycetous Yeasts: 4. The genome of Saccharomyces cerevisiae revisited  

Microsoft Academic Search

Since its completion more than 4 years ago, the sequence of Saccharomyces cerevisiae has been extensively used and studied. The original sequence has received a few corrections, and the identification of genes has been completed, thanks in particular to transcriptome analyses and to specialized studies on introns, tRNA genes, transposons or multigene families. In order to undertake the extensive comparative

Gaëlle Blandin; Pascal Durrens; Fredj Tekaia; Michel Aigle; Monique Bolotin-Fukuhara; Elisabeth Bon; Serge Casarégola; Jacky de Montigny; Claude Gaillardin; Andrée Lépingle; Bertrand Llorente; Alain Malpertuy; Cécile Neuvéglise; Odile Ozier-Kalogeropoulos; Arnaud Perrin; Serge Potier; Jean-Luc Souciet; Emmanuel Talla; Claire Toffano-Nioche; Micheline Wésolowski-Louvel; Christian Marck; Bernard Dujon



Biosynthesis of isoprenoids, polyunsaturated fatty acids and flavonoids in Saccharomyces cerevisiae  

Microsoft Academic Search

Industrial biotechnology employs the controlled use of microorganisms for the production of synthetic chemicals or simple biomass that can further be used in a diverse array of applications that span the pharmaceutical, chemical and nutraceutical industries. Recent advances in metagenomics and in the incorporation of entire biosynthetic pathways into Saccharomyces cerevisiae have greatly expanded both the fitness and the repertoire

Joseph A Chemler; Yajun Yan; Mattheos AG Koffas