WorldWideScience

Sample records for saccharomyces cerevisiae encodes

  1. Small toxic protein encoded on chromosome VII of Saccharomyces cerevisiae.

    Science.gov (United States)

    Makanae, Koji; Kintaka, Reiko; Ishikawa, Koji; Moriya, Hisao

    2015-01-01

    In a previous study, we found an unknown element that caused growth inhibition after its copy number increased in the 3' region of DIE2 in Saccharomyces cerevisiae. In this study, we further identified this element and observed that overexpression of a small protein (sORF2) of 57 amino acids encoded in this region caused growth inhibition. The transcriptional response and multicopy suppression of the growth inhibition caused by sORF2 overexpression suggest that sORF2 overexpression inhibits the ergosterol biosynthetic pathway. sORF2 was not required in the normal growth of S. cerevisiae, and not conserved in related yeast species including S. paradoxus. Thus, sORF2 (designated as OTO1) is an orphan ORF that determines the specificity of this species. PMID:25781884

  2. Two functional alpha-tubulin genes of the yeast Saccharomyces cerevisiae encode divergent proteins.

    OpenAIRE

    Schatz, P. J.; Pillus, L.; Grisafi, P.; Solomon, F.; Botstein, D.

    1986-01-01

    Two alpha-tubulin genes from the budding yeast Saccharomyces cerevisiae were identified and cloned by cross-species DNA homology. Nucleotide sequencing studies revealed that the two genes, named TUB1 and TUB3, encoded gene products of 447 and 445 amino acids, respectively, that are highly homologous to alpha-tubulins from other species. Comparison of the sequences of the two genes revealed a 19% divergence between the nucleotide sequences and a 10% divergence between the amino acid sequences....

  3. Regulation of pyc1 encoding pyruvate carboxylase isozyme I by nitrogen sources in Saccharomyces cerevisiae

    OpenAIRE

    Huet, Carine; Mene?ndez, Javier; Gancedo, Carlos; Franc?ois, Jean M.

    2000-01-01

    In Saccharomyces cerevisiae, the existence of PYC1 and PYC2 encoding cytosolic pyruvate carboxylase isoform I and II is rather puzzling, owing to the lack of potent differential gene regulation by the carbon sources. We report several findings indicating that these two genes are differentially regulated by the nature of the nitrogen source. In wild-type cells, the activity of pyruvate carboxylase, which is the sum of pyruvate carboxylase isoform I and II, was two- to fivefold lower in carbon ...

  4. GAC1 may encode a regulatory subunit for protein phosphatase type 1 in Saccharomyces cerevisiae.

    OpenAIRE

    Franc?ois, J. M.; Thompson-jaeger, S.; Skroch, J.; Zellenka, U.; Spevak, W.; Tatchell, K.

    1992-01-01

    Elevated dosage of the GAC1 gene from the yeast Saccharomyces cerevisiae causes hyperaccumulation of glycogen whereas a gene disruption of GAC1 results in reduced glycogen levels. Glycogen synthase is almost entirely in the active, glucose 6-phosphate-independent, form in cells with increased gene dosage of GAC1 whereas the enzyme is mostly in the inactive form in strains lacking GAC1. GAC1 encodes an 88 kDa protein that is similar to the regulatory subunit (RG1) of phosphoprotein phosphatase...

  5. Genes regulation encoding ADP/ATP carrier in yeasts Saccharomyces cerevisiae and Candida parapsilosis

    International Nuclear Information System (INIS)

    Genes encoding a mitochondrial ADP/ATP carrier (AAC) in yeast Saccharomyces cerevisiae and Candida parapsilosis were investigated. AAC2 is coding for the major AAC isoform in S. cerevisiae. We suggest that AAC2 is a member of a syn-expression group of genes encoding oxidative phosphorylation proteins. Within our previous studies on the regulation of the AAC2 transcription an UAS (-393/-268) was identified that is essential for the expression of this gene. Two functional regulatory cis-elements are located within this UAS -binding sites for an ABFl factor and for HAP2/3/4/5 heteromeric complex. We examined relative contributions and mutual interactions of the ABFl and HAP2/3/4/5 factors in the activation of transcription from the UAS of the AAC2 gene. The whole UAS was dissected into smaller sub-fragments and tested for (i) the ability to form DNA-protein complexes with cellular proteins in vitro, (ii) the ability to confer heterologous expression using AAC3 gene lacking its own promoter, and (iii) the expression of AAC3-lacZ fusion instead of intact AAC3 gene. The obtained results demonstrated that: a) The whole UAS as well as sub-fragment containing only ABF1-binding site are able to form DNA-protein complexes with cellular proteins in oxygen- and heme- dependent manner. The experiments with antibody against the ABF1 showed that the ABF1 factor is one of the proteins binding to AAC2 promoter. We have been unsuccessful to prove the binding of cellular proteins to the HAP2/3/4/5-binding site. However, the presence of HAP2/3/4/5-binding site is necessary to drive a binding of cellular proteins to the ABF1-binding site in carbon source-dependent manner. b) The presence of both ABF1- and HAP2/3/4/5-binding sites and original spacing between them is necessary to confer the growth of Aaac2 mutant strain on non- fermentable carbon source when put in front of AAC3 gene introduced on centromeric vector to Aaac2 mutant strain. c) For the activation of AAC3-lacZ expression on both fermentable and non-fermentable carbon sources the only presence of two copies of HAP2/3/4/5-binding site is sufficient. However, activation of AAC3-lacZ expression by two copies of HAP2/3/4/5-binding site is very low. We can conclude that the presence of both ABF1- and HAP2/3/4/5-binding sites and original spacing between them is necessary to get strong activation of AAC2 gene. A gene homologous to Saccharomyces cerevisiae AAC genes coding for mitochondrial ADP/ATP carriers has been cloned from pathogenic yeast Candida parapsilosis. The cloned gene was sequenced and found to encode a polypeptide of 303 amino acids that shows homology with other yeast and mammal mitochondrial ADP/ATP carriers. The gene was designed CpAAC1 and was able to complement the growth phenotype of S. cerevisiae double deletion mutant (?aac2?aac3). The expression of the CpAAC1 gene was affected at normal aerobic conditions by the nature of carbon source used for growth. The concentration of oxygen had no effect to the expression of this gene. Hybridization experiments indicate that C. parapsilosis possesses a single gene encoding a mitochondrial ADP/ ATP carrier. (author)

  6. The Lactate-Proton Symport of Saccharomyces cerevisiae Is Encoded by JEN1

    OpenAIRE

    Casal, Margarida; Paiva, Sandra; Andrade, Raquel P.; Gancedo, Carlos; Leão, Cecília

    1999-01-01

    A mutant of Saccharomyces cerevisiae deficient in the lactate-proton symport was isolated. Transformation of the mutant with a yeast genomic library allowed the isolation of the gene JEN1 that restored lactate transport. Disruption of JEN1 abolished uptake of lactate. The results indicate that, under the experimental conditions tested, no other monocarboxylate permease is able to efficiently transport lactate in S. cerevisiae.

  7. RAD6 gene of Saccharomyces cerevisiae encodes a protein containing a tract of 13 consecutive aspartates

    International Nuclear Information System (INIS)

    The RAD6 gene of Saccharomyces cerevisiae is required for postreplication repair of UV-damaged DNA, for induced mutagenesis, and for sporulation. The authors have mapped the transcripts and determined the nucleotide sequence of the cloned RAD6 gene. The RAD6 gene encodes two transcripts of 0.98 and 0.86 kilobases which differ only in their 3' termini. The transcribed region contains an open reading frame of 516 nucleotides. The rad6-1 and rad6-3 mutant alleles, which the authors have cloned and sequenced, introduce amber and ochre nonsense mutations, respectively into the open reading frame, proving that it encodes the RAD6 protein. The RAD6 protein predicted by the nucleotide sequence is 172 amino acids long, has a molecular weight of 19,704, and contains 23.3% acidic and 11.6% basic residues. Its most striking feature is the highly acidic carboxyl terminus: 20 of the 23 terminal amino acids are acidic, including 13 consecutive aspartates. RAD6 protein thus resembles high mobility group proteins HMG-1 and HMG-2, which each contain a carboxyl-proximal tract of acidic amino acids. 48 references, 6 figures

  8. GIT1, a gene encoding a novel transporter for glycerophosphoinositol in Saccharomyces cerevisiae.

    OpenAIRE

    Patton-Vogt, J L; Henry, S A

    1998-01-01

    Phosphatidylinositol catabolism in Saccharomyces cerevisiae cells cultured in media containing inositol results in the release of glycerophosphoinositol (GroPIns) into the medium. As the extracellular concentration of inositol decreases with growth, the released GroPIns is transported back into the cell. Exploiting the ability of the inositol auxotroph, ino1, to use exogenous GroPIns as an inositol source, we have isolated mutants (Git-) defective in the uptake and metabolism of GroPIns. One ...

  9. ERG1, encoding squalene epoxidase, is located on the right arm of chromosome VII of Saccharomyces cerevisiae.

    Science.gov (United States)

    Landl, K M; Klösch, B; Turnowsky, F

    1996-05-01

    The ERG1 gene of Saccharomyces cerevisiae encodes squalene epoxidase, a key enzyme in the ergosterol pathway. ERG1 is an essential gene. Disruption of the gene with URA3 results in a lethal phenotype when cells are grown under aerobic conditions, even in the presence of ergosterol. However, cells are viable in the presence of ergosterol under anaerobic growth conditions during which ergosterol is taken up by cells. Physical and genetic mapping data reveal that ERG1 is located on the right arm of chromosome VII proximal to QCR9 at a distance of 14.6 cM from ADE3. PMID:8771716

  10. Isolation of the GFA1 gene encoding glucosamine-6-phosphate synthase of Sporothrix schenckii and its expression in Saccharomyces cerevisiae.

    Science.gov (United States)

    Sánchez-López, Juan Francisco; González-Ibarra, Joaquín; Álvarez-Vargas, Aurelio; Milewski, Slawomir; Villagómez-Castro, Julio César; Cano-Canchola, Carmen; López-Romero, Everardo

    2015-06-01

    Glucosamine-6-phosphate synthase (GlcN-6-P synthase) is an essential enzyme involved in cell wall biogenesis that has been proposed as a strategic target for antifungal chemotherapy. Here we describe the cloning and functional characterization of Sporothrix schenckii GFA1 gene which was isolated from a genomic library of the fungus. The gene encodes a predicted protein of 708 amino acids that is homologous to GlcN-6-P synthases from other sources. The recombinant enzyme restored glucosamine prototrophy of the Saccharomyces cerevisiae gfa1 null mutant. Purification and biochemical analysis of the recombinant enzyme revealed some differences from the wild type enzyme, such as improved stability and less sensitivity to UDP-GlcNAc. The sensitivity of the recombinant enzyme to the selective inhibitor FMDP [N(3)-(4-methoxyfumaroyl)-l-2,3-diaminopropanoic acid] and other properties were similar to those previously reported for the wild type enzyme. PMID:25514203

  11. LIPID PHOSPHATE PHOSPHATASES FROM SACCHAROMYCES CEREVISIAE

    OpenAIRE

    Carman, George M.; Wu, Wen-I

    2007-01-01

    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 ...

  12. HFA1 encoding an organelle-specific acetyl-CoA carboxylase controls mitochondrial fatty acid synthesis in Saccharomyces cerevisiae.

    Science.gov (United States)

    Hoja, Ursula; Marthol, Sandra; Hofmann, Jörg; Stegner, Sabine; Schulz, Rainer; Meier, Sandra; Greiner, Eva; Schweizer, Eckhart

    2004-05-21

    The Saccharomyces cerevisiae gene, HFA1, encodes a >250-kDa protein, which is required for mitochondrial function. Hfa1p exhibits 72% overall sequence similarity (54% identity) to ACC1-encoded yeast cytoplasmic acetyl-CoA carboxylase. Nevertheless, HFA1 and ACC1 functions are not overlapping because mutants of the two genes have different phenotypes and do not complement each other. Whereas ACC1 is involved in cytoplasmic fatty acid synthesis, the phenotype of hfa1Delta disruptants resembles that of mitochondrial fatty-acid synthase mutants. They fail to grow on lactate or glycerol, and the mitochondrial cofactor, lipoic acid, is reduced to acetyl-CoA carboxylase. Expression of HFA1 under the control of the ACC1 promoter restored cellular ACC activity in ACC1-defective yeast mutants to wild type levels. From this finding, it is concluded that HFA1 encodes a specific mitochondrial acetyl-CoA carboxylase providing malonyl-CoA for intraorganellar fatty acid and, in particular, lipoic acid synthesis. PMID:14761959

  13. GUP1 and its close homologue GUP2, encoding multi-membrane-spanning proteins involved in active glycerol uptake in Saccharomyces cerevisiae

    OpenAIRE

    Holst, Bjørn; Lunde, Christina; Lages, Fernanda; Oliveira, Rui Pedro Soares; Lucas, Ca?ndida; Kielland-brandt, Morten

    2000-01-01

    Many yeast species can utilise glycerol, both as sole carbon source and as an osmolyte. In Saccharomyces cerevisiae, physiological studies have previously shown the presence of an active uptake system driven by electrogenic proton symport. We have used transposon mutagenesis to isolate mutants affected in the transport of glycerol into the cell. Here we present the identification of YGL084c, encoding a multi-membrane-spanning protein, as being essential for proton symport of glycerol into Sac...

  14. The GCR1 gene encodes a positive transcriptional regulator of the enolase and glyceraldehyde-3-phosphate dehydrogenase gene families in Saccharomyces cerevisiae.

    OpenAIRE

    Holland, M. J.; Yokoi, T.; Holland, J. P.; Myambo, K.; Innis, M. A.

    1987-01-01

    The intracellular concentrations of the polypeptides encoded by the two enolase (ENO1 and ENO2) and three glyceraldehyde-3-phosphate dehydrogenase (TDH1, TDH2, and TDH3) genes were coordinately reduced more than 20-fold in a Saccharomyces cerevisiae strain carrying the gcr1-1 mutation. The steady-state concentration of glyceraldehyde-3-phosphate dehydrogenase mRNA was shown to be approximately 50-fold reduced in the mutant strain. Overexpression of enolase and glyceraldehyde-3-phosphate dehyd...

  15. The Saccharomyces cerevisiae NPS1 gene, a novel CDC gene which encodes a 160 kDa nuclear protein involved in G2 phase control.

    OpenAIRE

    Tsuchiya, E.; Uno, M.; Kiguchi, A.; Masuoka, K.; Kanemori, Y.; Okabe, S.; Mikayawa, T.

    1992-01-01

    We have cloned the gene NPS1 (nuclear protein of Saccharomyces) which encodes a nuclear protein of mol. wt 156 735 Daltons (1359 amino acids) essential for cell growth. NPS1 contains a 2 kb sequence that is highly homologous to the S. cerevisiae SNF2/GAM1 gene known as a transcriptional regulator for multiple genes. However, the NPS1 gene was found to have a distinct function from SNF2/GAM1. The growth of the cells carrying a nps1 delta :: URA3 deletion allele and galactose-inducible NPS1 on ...

  16. Interactions of TLC1 (Which Encodes the RNA Subunit of Telomerase), TEL1, and MEC1 in Regulating Telomere Length in the Yeast Saccharomyces cerevisiae

    OpenAIRE

    Ritchie, Kim B.; Mallory, Julia C.; Petes, Thomas D.

    1999-01-01

    In the yeast Saccharomyces cerevisiae, chromosomes terminate with a repetitive sequence [poly(TG1–3)] 350 to 500 bp in length. Strains with a mutation of TEL1, a homolog of the human gene (ATM) mutated in patients with ataxia telangiectasia, have short but stable telomeric repeats. Mutations of TLC1 (encoding the RNA subunit of telomerase) result in strains that have continually shortening telomeres and a gradual loss of cell viability; survivors of senescence arise as a consequence of a Ra...

  17. Cloning and bacterial expression of the CYS3 gene encoding cystathionine gamma-lyase of Saccharomyces cerevisiae and the physicochemical and enzymatic properties of the protein.

    OpenAIRE

    S. Yamagata; D'Andrea, R J; Fujisaki, S.; Isaji, M.; Nakamura, K

    1993-01-01

    By screening a yeast genomic library, we isolated and characterized a gene rescuing the cysteine requirement in a "cys1" strain of Saccharomyces cerevisiae. Except for four residues in the open reading frame composed of 1,182 nucleotides, the DNA sequence was the same as that for the CYS3 (CYI1) gene, encoding cystathionine gamma-lyase (EC 4.4.1.1), and isolated previously as a cycloheximide-induced gene (B. Ono, K. Tanaka, K. Naito, C. Heike, S. Shinoda, S. Yamamoto, S. Ohmori, T. Oshima, an...

  18. Structure and function of the Saccharomyces cerevisiae CDC2 gene encoding the large subunit of DNA polymerase III.

    OpenAIRE

    Boulet, A.; Simon, M.; Faye, G.; Bauer, G. A.; Burgers, P. M.

    1989-01-01

    Saccharomyces cerevisiae cdc2 mutants arrest in the S-phase of the cell cycle when grown at the non-permissive temperature, implicating this gene product as essential for DNA synthesis. The CDC2 gene has been cloned from a yeast genomic library in vector YEp13 by complementation of a cdc2 mutation. An open reading frame coding for a 1093 amino acid long protein with a calculated mol. wt of 124,518 was determined from the sequence. This putative protein shows significant homology with a class ...

  19. Purification and Properties of an Esterase from the Yeast Saccharomyces cerevisiae and Identification of the Encoding Gene

    OpenAIRE

    Degrassi, Giuliano; Uotila, Lasse; Klima, Raffaella; Venturi, Vittorio

    1999-01-01

    We purified an intracellular esterase that can function as an S-formylglutathione hydrolase from the yeast Saccharomyces cerevisiae. Its molecular mass was 40 kDa, as determined by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point was 5.0 by isoelectric focusing. The enzyme activity was optimal at 50°C and pH 7.0. The corresponding gene, YJLO68C, was identified by its N-terminal amino acid sequence and is not essential for cell viability. Nul...

  20. Saccharomyces cerevisiae YOR071C encodes the high affinity nicotinamide riboside transporter Nrt1.

    Science.gov (United States)

    Belenky, Peter A; Moga, Tiberiu G; Brenner, Charles

    2008-03-28

    NAD(+) is an essential coenzyme for hydride transfer enzymes and a substrate of sirtuins and other NAD(+)-consuming enzymes. Nicotinamide riboside is a recently discovered eukaryotic NAD(+) precursor converted to NAD(+) via the nicotinamide riboside kinase pathway and by nucleosidase activity and nicotinamide salvage. Nicotinamide riboside supplementation of yeast extends replicative life span on high glucose medium. The molecular basis for nicotinamide riboside uptake was unknown in any eukaryote. Here, we show that deletion of a single gene, YOR071C, abrogates nicotinamide riboside uptake without altering nicotinic acid or nicotinamide import. The gene, which is negatively regulated by Sum1, Hst1, and Rfm1, fully restores nicotinamide riboside import and utilization when resupplied to mutant yeast cells. The encoded polypeptide, Nrt1, is a predicted deca-spanning membrane protein related to the thiamine transporter, which functions as a pH-dependent facilitator with a K(m) for nicotinamide riboside of 22 microm. Nrt1-related molecules are conserved in particular fungi, suggesting a similar basis for nicotinamide riboside uptake. PMID:18258590

  1. Phosphoenolpyruvate Carboxykinase as the Sole Anaplerotic Enzyme in Saccharomyces cerevisiae?

    OpenAIRE

    Zelle, R.M.; Trueheart, J; Harrison, J C; Pronk, J. T.; van Maris, A J A

    2010-01-01

    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 ...

  2. Glucose Signaling in Saccharomyces cerevisiae

    OpenAIRE

    Santangelo, George M.

    2006-01-01

    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...

  3. Proteomics of Saccharomyces cerevisiae Organelles*

    OpenAIRE

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

    2009-01-01

    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 organe...

  4. The mutant type 1 protein phosphatase encoded by glc7-1 from Saccharomyces cerevisiae fails to interact productively with the GAC1-encoded regulatory subunit.

    OpenAIRE

    Stuart, J. S.; Frederick, D. L.; Varner, C. M.; Tatchell, K.

    1994-01-01

    Loss-of-function gac1 mutants of Saccharomyces cerevisiae fail to accumulate normal levels of glycogen because of low glycogen synthase activity. Increased dosage of GAC1 results in increased activity of glycogen synthase and a corresponding hyperaccumulation of glycogen. The glycogen accumulation phenotype of gac1 is similar to that of glc7-1, a type 1 protein phosphatase mutant. We have partially characterized the GAC1 gene product (Gac1p) and show that levels of Gac1p increase during growt...

  5. Virus-like particle capsid proteins encoded by different L double-stranded RNAs of Saccharomyces cerevisiae: their roles in maintenance of M double-stranded killer plasmids.

    OpenAIRE

    El-Sherbeini, M; Tipper, D J; Mitchell, D J; Bostian, K. A.

    1984-01-01

    The plasmid determinants of killer phenotypes in type K1 and K2 killer yeast cells are the 1.9-kilobase (kb) M1 and 1.7-kb M2 double-stranded RNAs (dsRNAs), respectively. These are dependent for their maintenance and encapsidation, in Saccharomyces cerevisiae virus ScV-M1 or ScV-M2 virus-like particles, on the capsid provided by one of a group of moderately related 4.7-kb dsRNAs called LA. The L1A and L2A dsRNAs found in naturally isolated K1 and K2 killers encode 88-kilodalton VL1A-P1 and 86...

  6. Allantoin transport in Saccharomyces cerevisiae.

    Science.gov (United States)

    Sumrada, R; Cooper, T G

    1977-01-01

    Allantoin uptake in both growing and resting cultures of Saccharomyces cerevisiae occurs by a low-Km (ca. 15 micrometer) transport system that uses energy that is likely generated in the cytoplasm. This conclusion was based on the observation that transport did not occur in the absence of glucose or the presence of dinitrophenol, carbonyl cyanide-m-chloro-phenyl hydrazine, fluoride, or arsenate ions. Normal uptake was observed, however, in the presence of cyanide. The rate of accumulation was maximal at pH 5.2. In contrast to the urea transport system, allantoin uptake appeared to be unidirectional. Preloaded, radioactive allantoin was not lost from cells suspended in allantoin-free buffer and did not exchange with exogenously added, nonradioactive allantoin. Treatment of preloaded cells with nystatin, however, released the accumulated radioactivity. Allantoin accumulated within cells was isolated and shown to be chemically unaltered. PMID:19421

  7. Progress in Metabolic Engineering of Saccharomyces cerevisiae

    OpenAIRE

    Nevoigt, Elke

    2008-01-01

    Summary: The traditional use of the yeast Saccharomyces cerevisiae in alcoholic fermentation has, over time, resulted in substantial accumulated knowledge concerning genetics, physiology, and biochemistry as well as genetic engineering and fermentation technologies. S. cerevisiae has become a platform organism for developing metabolic engineering strategies, methods, and tools. The current review discusses the relevance of several engineering strategies, such as rational and inverse metabolic...

  8. Fatal Saccharomyces Cerevisiae Aortic Graft Infection

    Science.gov (United States)

    Meyer, Michael (Technical Monitor); Smith, Davey; Metzgar, David; Wills, Christopher; Fierer, Joshua

    2002-01-01

    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. We present a case of S. cerevisiae fungemia and aortic graft infection in an immunocompetent adult. This is the first reported case of S. cerevisiue fungemia where the identity of the pathogen was confirmed by rRNA sequencing.

  9. Transcriptional Regulatory Networks in Saccharomyces cerevisiae

    Science.gov (United States)

    Lee, Tong Ihn; Rinaldi, Nicola J.; Robert, François; Odom, Duncan T.; Bar-Joseph, Ziv; Gerber, Georg K.; Hannett, Nancy M.; Harbison, Christopher T.; Thompson, Craig M.; Simon, Itamar; Zeitlinger, Julia; Jennings, Ezra G.; Murray, Heather L.; Gordon, D. Benjamin; Ren, Bing; Wyrick, John J.; Tagne, Jean-Bosco; Volkert, Thomas L.; Fraenkel, Ernest; Gifford, David K.; Young, Richard A.

    2002-10-01

    We have determined how most of the transcriptional regulators encoded in the eukaryote Saccharomyces cerevisiae associate with genes across the genome in living cells. Just as maps of metabolic networks describe the potential pathways that may be used by a cell to accomplish metabolic processes, this network of regulator-gene interactions describes potential pathways yeast cells can use to regulate global gene expression programs. We use this information to identify network motifs, the simplest units of network architecture, and demonstrate that an automated process can use motifs to assemble a transcriptional regulatory network structure. Our results reveal that eukaryotic cellular functions are highly connected through networks of transcriptional regulators that regulate other transcriptional regulators.

  10. mRNAs encoding polarity and exocytosis factors are cotransported with the cortical endoplasmic reticulum to the incipient bud in Saccharomyces cerevisiae.

    Science.gov (United States)

    Aronov, Stella; Gelin-Licht, Rita; Zipor, Gadi; Haim, Liora; Safran, Einat; Gerst, Jeffrey E

    2007-05-01

    Polarized growth in the budding yeast Saccharomyces cerevisiae depends upon the asymmetric localization and enrichment of polarity and secretion factors at the membrane prior to budding. We examined how these factors (i.e., Cdc42, Sec4, and Sro7) reach the bud site and found that their respective mRNAs localize to the tip of the incipient bud prior to nuclear division. Asymmetric mRNA localization depends upon factors that facilitate ASH1 mRNA localization (e.g., the 3' untranslated region, She proteins 1 to 5, Puf6, actin cytoskeleton, and a physical association with She2). mRNA placement precedes protein enrichment and subsequent bud emergence, implying that mRNA localization contributes to polarization. Correspondingly, mRNAs encoding proteins which are not asymmetrically distributed (i.e., Snc1, Mso1, Tub1, Pex3, and Oxa1) are not polarized. Finally, mutations which affect cortical endoplasmic reticulum (ER) entry and anchoring in the bud (myo4Delta, sec3Delta, and srp101) also affect asymmetric mRNA localization. Bud-localized mRNAs, including ASH1, were found to cofractionate with ER microsomes in a She2- and Sec3-dependent manner; thus, asymmetric mRNA transport and cortical ER inheritance are connected processes in yeast. PMID:17339339

  11. A cell cycle-responsive transcriptional control element and a negative control element in the gene encoding DNA polymerase alpha in Saccharomyces cerevisiae.

    Science.gov (United States)

    Gordon, C B; Campbell, J L

    1991-01-01

    Transcription of the POL1 gene of Saccharomyces cerevisiae, which encodes DNA polymerase alpha, the DNA polymerase required for the initiation of DNA replication, has previously been shown to be cell cycle regulated. To understand how the POL1 gene senses cell cycle position, we have investigated the cis-acting elements that respond to the factors that govern cell cycle progression. In this report we demonstrate that a region of 54 nucleotides containing the repeated element ACGCGT, which conforms to an Mlu I restriction endonuclease recognition site, contains all information necessary for transcriptional activation and cell cycle responsiveness. Although oligonucleotides lacking either one or both of the repeated Mlu I sites can function as an upstream activating sequence, the presence of at least one Mlu I site stimulates expression and, moreover, is absolutely essential for cell cycle regulation. A synthetic oligonucleotide corresponding to a 19-base-pair sequence in the POL1 promoter containing one Mlu I site can function as an autonomous cell cycle-responsive upstream element (upstream activation sequence) with temporal regulation indistinguishable from that previously described for the POL1 gene. Thus, the Mlu I site is an essential part of a cis-acting element responsible for the observed periodic activation. This sequence differs from previously defined cell cycle-responsive transcriptional control elements in the yeast HO endonuclease and histone genes. We also present evidence for a negative regulatory element in the 5' flanking region of the Mlu I upstream activation sequence. Images PMID:2068085

  12. Cloning and bacterial expression of the CYS3 gene encoding cystathionine gamma-lyase of Saccharomyces cerevisiae and the physicochemical and enzymatic properties of the protein.

    Science.gov (United States)

    Yamagata, S; D'Andrea, R J; Fujisaki, S; Isaji, M; Nakamura, K

    1993-08-01

    By screening a yeast genomic library, we isolated and characterized a gene rescuing the cysteine requirement in a "cys1" strain of Saccharomyces cerevisiae. Except for four residues in the open reading frame composed of 1,182 nucleotides, the DNA sequence was the same as that for the CYS3 (CYI1) gene, encoding cystathionine gamma-lyase (EC 4.4.1.1), and isolated previously as a cycloheximide-induced gene (B. Ono, K. Tanaka, K. Naito, C. Heike, S. Shinoda, S. Yamamoto, S. Ohmori, T. Oshima, and A. Toh-e, J. Bacteriol. 174:pp.3339-3347, 1992). S. cerevisiae "cys1" strains carry two closely linked mutations; one (cys1) causes a defect in serine O-acetyltransferase (EC 2.3.1.30), and another, designated cys3, impairs cystathionine gamma-lyase activity. Rescue of the cysteine requirement by the gene encoding cystathionine gamma-lyase is consistent with both defects being responsible for the cysteine auxotrophy. In an effort to further determine the physicochemical and enzymatic properties of this enzyme, a coding fragment was cloned into an Escherichia coli expression plasmid, and the protein was produced in the bacteria. The induced protein was extracted by sonication and purified to homogeneity through one course of DEAE-cellulose column chromatography. The yield of the protein was approximately 150 mg from cells cultured in 1 liter of L broth. The protein showed molecular weights of approximately 194,000 and 48,000 (for the subunit), suggesting a tetrameric structure. An s20,w value of 8.8 was estimated by centrifugation in a sucrose concentration gradient. No sulfhydryl groups were detected, which is consistent with the absence of cysteine residues in the coding sequence. The isoelectric point was at pH 5.2. The protein showed a number of cystathionine-related activities, i.e., cystathionine beta-lyase (EC 4.4.1.8), cystathionine gamma-lyase, and cystathionine gamma-synthase (EC 4.2.99.9) with L-homoserine as substrate. In addition, we demonstrated L-homoserine sulfhydrylase (adding H2S) activity but could find no detectable serine O-acteyltransferease activity. In this paper, we compare the enzymatic properties of the protein with those of homologous enzymes previously reported and discuss the possibility that this enzyme has a physiological role as cystathionine Beta-lyase and cystathionine gamma-synthase in addition to its previously described role as cystathionine gamma-lyase. PMID:8335636

  13. 40 CFR 180.1246 - Yeast Extract Hydrolysate from Saccharomyces cerevisiae: exemption from the requirement of a...

    Science.gov (United States)

    2010-07-01

    ...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...

  14. The Saccharomyces cerevisiae aquaporin Aqy1 is involved in sporulation

    OpenAIRE

    Sidoux-walter, Fre?de?ric; Pettersson, Nina; Hohmann, Stefan

    2004-01-01

    Aquaporins mediate rapid selective water transport across biological membranes. Elucidation of their precise physiological roles promises important insight into cellular and organismal osmoregulation. The genome of the yeast Saccharomyces cerevisiae encodes two similar but differentially regulated aquaporins. Here, we show that expression of AQY1 is stimulated during sporulation and that the Aqy1 protein is detectable exclusively in spore membranes. When spores are rapidly frozen, those that ...

  15. Functional studies of aldo-keto reductases in Saccharomyces cerevisiae*

    OpenAIRE

    CHANG Qing; Griest, Terry A.; Harter, Theresa M.; PETRASH, J. MARK

    2006-01-01

    We utilized the budding yeast Saccharomyces cerevisiae as a model to systematically explore physiological roles for yeast and mammalian aldo-keto reductases. Six open reading frames encoding putative aldo-keto reductases were identified when the yeast genome was queried against the sequence for human aldose reductase, the prototypical mammalian aldo-keto reductase. Recombinant proteins produced from five of these yeast open reading frames demonstrated NADPH-dependent reductase activity with a...

  16. Metabolic Engineering of Glycerol Production in Saccharomyces cerevisiae:

    OpenAIRE

    Overkamp, K. M.; Bakker, B. M.; Kotter, P.; Luttik, M. A. H.; Dijken, J. P.; Pronk, J. T.

    2002-01-01

    Inactivation of TPI1, the Saccharomyces cerevisiae structural gene encoding triose phosphate isomerase, completely eliminates growth on glucose as the sole carbon source. In tpi1-null mutants, intracellular accumulation of dihydroxyacetone phosphate might be prevented if the cytosolic NADH generated in glycolysis by glyceraldehyde-3-phosphate dehydrogenase were quantitatively used to reduce dihydroxyacetone phosphate to glycerol. We hypothesize that the growth defect of tpi1-null mutants is c...

  17. Overproduction of Geranylgeraniol by Metabolically Engineered Saccharomyces cerevisiae?

    OpenAIRE

    Tokuhiro, Kenro; Muramatsu, Masayoshi; Ohto, Chikara; Kawaguchi, Toshiya; Obata, Shusei; Muramoto, Nobuhiko; Hirai, Masana; Takahashi, Haruo; Kondo, Akihiko; Sakuradani, Eiji; Shimizu, Sakayu

    2009-01-01

    (E, E, E)-Geranylgeraniol (GGOH) is a valuable starting material for perfumes and pharmaceutical products. In the yeast Saccharomyces cerevisiae, GGOH is synthesized from the end products of the mevalonate pathway through the sequential reactions of farnesyl diphosphate synthetase (encoded by the ERG20 gene), geranylgeranyl diphosphate synthase (the BTS1 gene), and some endogenous phosphatases. We demonstrated that overexpression of the diacylglycerol diphosphate phosphatase (DPP1) gene could...

  18. Metabolic Impact of Increased NADH Availability in Saccharomyces cerevisiae?

    OpenAIRE

    Hou, Jin; Scalcinati, Gionata; Oldiges, Marco; Vemuri, Goutham N.

    2009-01-01

    Engineering the level of metabolic cofactors to manipulate metabolic flux is emerging as an attractive strategy for bioprocess applications. We present the metabolic consequences of increasing NADH in the cytosol and the mitochondria of Saccharomyces cerevisiae. In a strain that was disabled in formate metabolism, we either overexpressed the native NAD+-dependent formate dehydrogenase in the cytosol or directed it into the mitochondria by fusing it with the mitochondrial signal sequence encod...

  19. Ferrofluid modified Saccharomyces cerevisiae cells for biocatalysis.

    Czech Academy of Sciences Publication Activity Database

    Šafa?íková, Miroslava; Mad?rová, Zde?ka; Šafa?ík, Ivo

    2009-01-01

    Ro?. 42, - (2009), s. 521-524. ISSN 0963-9969 R&D Projects: GA MPO 2A-1TP1/094; GA MŠk(CZ) OC 157 Institutional research plan: CEZ:AV0Z60870520 Keywords : Saccharomyces cerevisiae * magnetic fluid * hydrogen peroxide Subject RIV: EI - Biotechnology ; Bionics Impact factor: 2.414, year: 2009

  20. Tangential Ultrafiltration of Aqueous "Saccharomyces Cerevisiae" Suspensions

    Science.gov (United States)

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

    2008-01-01

    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…

  1. Mechanisms of Ethanol Tolerance in Saccharomyces cerevisiae

    Science.gov (United States)

    Saccharomyces cerevisiae is a superb ethanol producer, yet is also sensitive to higher ethanol concentrations especially under high gravity or very high gravity fermentation conditions. Ethanol tolerance is associated with interplay of complex networks at the genome level. Although significant eff...

  2. Global regulation of mitochondrial biogenesis in Saccharomyces cerevisiae: ABF1 and CPF1 play opposite roles in regulating expression of the QCR8 gene, which encodes subunit VIII of the mitochondrial ubiquinol-cytochrome c oxidoreductase.

    OpenAIRE

    Winde, J.H. de; Grivell, L A

    1992-01-01

    The multifunctional DNA-binding proteins ABF1 and CPF1 bind in a mutually exclusive manner to the promoter region of the QCR8 gene, which encodes 11-kDa subunit VIII of the Saccharomyces cerevisiae mitochondrial ubiquinol-cytochrome c oxidoreductase (QCR). We investigated the roles that the two factors play in transcriptional regulation of this gene. To this end, the overlapping binding sites for ABF1 and CPF1 were mutated and placed in the chromosomal context of the QCR8 promoter. The effect...

  3. A family of genes encode the multiple forms of the Saccharomyces cerevisiae ribosomal proteins equivalent to the Escherichia coli L12 protein and a single form of the L10-equivalent ribosomal protein.

    OpenAIRE

    Newton, C. H.; Shimmin, L. C.; Yee, J.; Dennis, P. P.

    1990-01-01

    The budding yeast Saccharomyces cerevisiae contains a family of genes that encodes four different but related small acidic ribosomal proteins designated L12eIA, L12eIB, L12eIIA, and L12eIIB and a single larger protein designated L10e. These proteins are equivalent (e) to the L12 and L10 proteins of Escherichia coli that assemble as a 4:1 complex onto the large ribosomal subunit. The five yeast genes (or their cDNAs) have been cloned and sequenced (M. Remacha, M. T. Saenz-Robles, M. D. Vilella...

  4. Cloning and characterization of the low-affinity cyclic AMP phosphodiesterase gene of Saccharomyces cerevisiae.

    OpenAIRE

    Nikawa, J.; Sass, P.; Wigler, M.

    1987-01-01

    Saccharomyces cerevisiae contains two genes which encode cyclic AMP (cAMP) phosphodiesterase. We previously isolated and characterized PDE2, which encodes a high-affinity cAMP phosphodiesterase. We have now isolated the PDE1 gene of S. cerevisiae, which encodes a low-affinity cAMP phosphodiesterase. These two genes represent highly divergent branches in the evolution of phosphodiesterases. High-copy-number plasmids containing either PDE1 or PDE2 can reverse the growth arrest defects of yeast ...

  5. mRNAs Encoding Polarity and Exocytosis Factors Are Cotransported with the Cortical Endoplasmic Reticulum to the Incipient Bud in Saccharomyces cerevisiae? †

    OpenAIRE

    Aronov, Stella; Gelin-licht, Rita; Zipor, Gadi; Haim, Liora; Safran, Einat; Gerst, Jeffrey E.

    2007-01-01

    Polarized growth in the budding yeast Saccharomyces cerevisiae depends upon the asymmetric localization and enrichment of polarity and secretion factors at the membrane prior to budding. We examined how these factors (i.e., Cdc42, Sec4, and Sro7) reach the bud site and found that their respective mRNAs localize to the tip of the incipient bud prior to nuclear division. Asymmetric mRNA localization depends upon factors that facilitate ASH1 mRNA localization (e.g., the 3? untranslated region,...

  6. Phosphate transport and sensing in Saccharomyces cerevisiae.

    OpenAIRE

    Wykoff, D D; O'Shea, E K

    2001-01-01

    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 starvat...

  7. Cell Wall Assembly in Saccharomyces cerevisiae

    OpenAIRE

    Lesage, Guillaume; Bussey, Howard

    2006-01-01

    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 i...

  8. Transformation of Saccharomyces cerevisiae by electroporation.

    OpenAIRE

    Delorme, E.

    1989-01-01

    A method for introducing heterologous DNA into Saccharomyces cerevisiae rapidly and efficiently by electroporation was developed. Transformant colonies appeared somewhat sooner than by the LiCl or spheroplast transformation method, and the time spent in manipulation was much less than for these two methods. The pores in the cell membrane formed by the high voltage of electroporation were resealed within 6 to 7 min after electroporation. At a capacitance of 25 microF, the optimum voltage was 2...

  9. Acid excreting mutants of yeast Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Saccharomyces cerevisiae mutants acidifying glucose medium containing bromocresol purple were shown to excrete protons when placed in unbuffered water in the absence of any external carbon source. The mutants belong to 16 different complementation groups. Most of them do not grow on glycerol and the excreted protons are associated to particular sets of organic anions such as citrate, aconitate, succinate, fumarate or malate. These novel types of respiratory mutations seem to be located in genes operating in the Krebs or glyoxylate cycle

  10. Improved Anaerobic Use of Arginine by Saccharomyces cerevisiae

    Science.gov (United States)

    Martin, Olga; Brandriss, Marjorie C.; Schneider, Gisbert; Bakalinsky, Alan T.

    2003-01-01

    Anaerobic arginine catabolism in Saccharomyces cerevisiae was genetically modified to allow assimilation of all four rather than just three of the nitrogen atoms in arginine. This was accomplished by bypassing normal formation of proline, an unusable nitrogen source in the absence of oxygen, and causing formation of glutamate instead. A pro3 ure2 strain expressing a PGK1 promoter-driven PUT2 allele encoding ?1-pyrroline-5-carboxylate dehydrogenase lacking a mitochondrial targeting sequence produced significant cytoplasmic activity, accumulated twice as much intracellular glutamate, and produced twice as much cell mass as the parent when grown anaerobically on limiting arginine as sole nitrogen source. PMID:12620851

  11. Novel Roles for Saccharomyces cerevisiae Mitotic Spindle Motors

    OpenAIRE

    Cottingham, Frank R.; Gheber, Larisa; Miller, Dana L.; Hoyt, M. Andrew

    1999-01-01

    The single cytoplasmic dynein and five of the six kinesin-related proteins encoded by Saccharomyces cerevisiae participate in mitotic spindle function. Some of the motors operate within the nucleus to assemble and elongate the bipolar spindle. Others operate on the cytoplasmic microtubules to effect spindle and nuclear positioning within the cell. This study reveals that kinesin-related Kar3p and Kip3p are unique in that they perform roles both inside and outside the nucleus. Kar3p, like Kip3...

  12. YNL134C from Saccharomyces cerevisiae encodes a novel protein with aldehyde reductase activity for detoxification of furfural derived from lignocellulosic biomass.

    Science.gov (United States)

    Zhao, Xianxian; Tang, Juan; Wang, Xu; Yang, Ruoheng; Zhang, Xiaoping; Gu, Yunfu; Li, Xi; Ma, Menggen

    2015-05-01

    Furfural and 5-hydroxymethylfurfural (HMF) are the two main aldehyde compounds derived from pentoses and hexoses, respectively, during lignocellulosic biomass pretreatment. These two compounds inhibit microbial growth and interfere with subsequent alcohol fermentation. Saccharomyces cerevisiae has the in situ ability to detoxify furfural and HMF to the less toxic 2-furanmethanol (FM) and furan-2,5-dimethanol (FDM), respectively. Herein, we report that an uncharacterized gene, YNL134C, was highly up-regulated under furfural or HMF stress and Yap1p and Msn2/4p transcription factors likely controlled its up-regulated expression. Enzyme activity assays showed that YNL134C is an NADH-dependent aldehyde reductase, which plays a role in detoxification of furfural to FM. However, no NADH- or NADPH-dependent enzyme activity was observed for detoxification of HMF to FDM. This enzyme did not catalyse the reverse reaction of FM to furfural or FDM to HMF. Further studies showed that YNL134C is a broad-substrate aldehyde reductase, which can reduce multiple aldehydes to their corresponding alcohols. Although YNL134C is grouped into the quinone oxidoreductase family, no quinone reductase activity was observed using 1,2-naphthoquinone or 9,10-phenanthrenequinone as a substrate, and phylogenetic analysis indicates that it is genetically distant to quinone reductases. Proteins similar to YNL134C in sequence from S. cerevisiae and other microorganisms were phylogenetically analysed. Copyright © 2015 John Wiley & Sons, Ltd. PMID:25656244

  13. Saccharomyces cerevisiae engineered for xylose metabolism exhibits a respiratory response.

    Science.gov (United States)

    Jin, Yong-Su; Laplaza, Jose M; Jeffries, Thomas W

    2004-11-01

    Native strains of Saccharomyces cerevisiae do not assimilate xylose. S. cerevisiae engineered for d-xylose utilization through the heterologous expression of genes for aldose reductase (XYL1), xylitol dehydrogenase (XYL2), and d-xylulokinase (XYL3 or XKS1) produce only limited amounts of ethanol in xylose medium. In recombinant S. cerevisiae expressing XYL1, XYL2, and XYL3, mRNA transcript levels for glycolytic, fermentative, and pentose phosphate enzymes did not change significantly on glucose or xylose under aeration or oxygen limitation. However, expression of genes encoding the tricarboxylic acid cycle, respiration enzymes (HXK1, ADH2, COX13, NDI1, and NDE1), and regulatory proteins (HAP4 and MTH1) increased significantly when cells were cultivated on xylose, and the genes for respiration were even more elevated under oxygen limitation. These results suggest that recombinant S. cerevisiae does not recognize xylose as a fermentable carbon source and that respiratory proteins are induced in response to cytosolic redox imbalance; however, lower sugar uptake and growth rates on xylose might also induce transcripts for respiration. A petite respiration-deficient mutant (rho degrees ) of the engineered strain produced more ethanol and accumulated less xylitol from xylose. It formed characteristic colonies on glucose, but it did not grow on xylose. These results are consistent with the higher respiratory activity of recombinant S. cerevisiae when growing on xylose and with its inability to grow on xylose under anaerobic conditions. PMID:15528549

  14. Myo-inositol transport in Saccharomyces cerevisiae.

    OpenAIRE

    Nikawa, J; Nagumo, T; Yamashita, S.

    1982-01-01

    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...

  15. [Engineering Saccharomyces cerevisiae for sclareol production].

    Science.gov (United States)

    Yang, Wei; Zhou, Yongjin; Liu, Wujun; Shen, Hongwei; Zhao, Zongbao K

    2013-08-01

    Sclareol is a member of labdane type diterpenes mostly used as fragrance ingredient. To enable microbial production of sclareol, synthetic pathways were constructed by incorporating labdenediol diphosphate synthase (LPPS) and terpene synthase (TPS) of the plant Salvia sclarea into Saccharomyces cerevisiae. It was found that sclareol production could be benefited by overexpression of key enzyme for precursor biosynthesis, construction of fusion protein for substrate channeling, and removal of signal peptides from LPPS and TPS. Under optimal shake flask culture conditions, strain S6 produced 8.96 mg/L sclareol. These results provided useful information for development of heterologous hosts for production of terpenoids. PMID:24364354

  16. Biosorption of americium-241 by Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    The biosorption of radionuclide 241Am from solution by Saccharomyces cerevisiae (S. cerevisiae), and the effects of experimental conditions on the adsorption were investigated. The preliminary results showed that S. cerevisiae is a very efficient biosorbent. An average of more than 99% of the total 241Am could be removed by S. cerevisiae of 2.1 g/l (dry weight) from 241Am solutions of 17.54-4386.0 ?g/l (2.22 MBq/l-555 MBq/l) with adsorption capacities of 7.45-1880.0 ?g/g biomass (dry weight) (0.94 MBq/g-237.9 MBq/g). The adsorption equilibrium was achieved within 1 hour and the optimum pH ranged 1-3. No significant differences on 241Am adsorption were observed at 10-45 deg C, or in solutions containing Au3+ or Ag+, even 2000 times above 241Am concentration. The relationship between concentrations and adsorption capacities of 241Am indicated the biosorption process should be described by the Freundlich adsorption isotherm. (author)

  17. Anaplerotic Role for Cytosolic Malic Enzyme in Engineered Saccharomyces cerevisiae Strains?

    OpenAIRE

    Zelle, R. M.; Harrison, J. C.; Pronk, J. T.; Maris, A. J. A.

    2010-01-01

    Malic enzyme catalyzes the reversible oxidative decarboxylation of malate to pyruvate and CO2. The Saccharomyces cerevisiae MAE1 gene encodes a mitochondrial malic enzyme whose proposed physiological roles are related to the oxidative, malate-decarboxylating reaction. Hitherto, the inability of pyruvate carboxylase-negative (Pyc–) S. cerevisiae strains to grow on glucose suggested that Mae1p cannot act as a pyruvate-carboxylating, anaplerotic enzyme. In this study, relocation of malic enzym...

  18. Characterization of fungal RTG2 genes in retrograde signaling of Saccharomyces cerevisiae

    OpenAIRE

    U?nlu?, Ercan Selc?uk; Narayanan, Lakshmi; Gordon, Donna M.

    2013-01-01

    Changes in the functional status of mitochondria result in the transcriptional activation of a subset of nuclear-encoded genes in a process referred to as retrograde signaling. In Saccharomyces cerevisiae, this molecular link between mitochondria and the nuclear genome is controlled by three key signaling proteins: Rtg1p, Rtg2p, and Rtg3p. Although the retrograde signaling response has been well characterized in S. cerevisiae, very little is known about this pathway in other fungi. In this st...

  19. PRIMARY STRUCTURE OF THE P450 LANOSTEROL DEMETHYLASE GENE FROM SACCHAROMYCES CEREVISIAE

    Science.gov (United States)

    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...

  20. Synthesis of Morphinan Alkaloids in Saccharomyces cerevisiae.

    Science.gov (United States)

    Fossati, Elena; Narcross, Lauren; Ekins, Andrew; Falgueyret, Jean-Pierre; Martin, Vincent J J

    2015-01-01

    Morphinan alkaloids are the most powerful narcotic analgesics currently used to treat moderate to severe and chronic pain. The feasibility of morphinan synthesis in recombinant Saccharomyces cerevisiae starting from the precursor (R,S)-norlaudanosoline was investigated. Chiral analysis of the reticuline produced by the expression of opium poppy methyltransferases showed strict enantioselectivity for (S)-reticuline starting from (R,S)-norlaudanosoline. In addition, the P. somniferum enzymes salutaridine synthase (PsSAS), salutaridine reductase (PsSAR) and salutaridinol acetyltransferase (PsSAT) were functionally co-expressed in S. cerevisiae and optimization of the pH conditions allowed for productive spontaneous rearrangement of salutaridinol-7-O-acetate and synthesis of thebaine from (R)-reticuline. Finally, we reconstituted a 7-gene pathway for the production of codeine and morphine from (R)-reticuline. Yeast cell feeding assays using (R)-reticuline, salutaridine or codeine as substrates showed that all enzymes were functionally co-expressed in yeast and that activity of salutaridine reductase and codeine-O-demethylase likely limit flux to morphine synthesis. The results of this study describe a significant advance for the synthesis of morphinans in S. cerevisiae and pave the way for their complete synthesis in recombinant microbes. PMID:25905794

  1. Synthesis of Morphinan Alkaloids in Saccharomyces cerevisiae

    Science.gov (United States)

    Fossati, Elena; Narcross, Lauren; Ekins, Andrew; Falgueyret, Jean-Pierre; Martin, Vincent J. J.

    2015-01-01

    Morphinan alkaloids are the most powerful narcotic analgesics currently used to treat moderate to severe and chronic pain. The feasibility of morphinan synthesis in recombinant Saccharomyces cerevisiae starting from the precursor (R,S)-norlaudanosoline was investigated. Chiral analysis of the reticuline produced by the expression of opium poppy methyltransferases showed strict enantioselectivity for (S)-reticuline starting from (R,S)-norlaudanosoline. In addition, the P. somniferum enzymes salutaridine synthase (PsSAS), salutaridine reductase (PsSAR) and salutaridinol acetyltransferase (PsSAT) were functionally co-expressed in S. cerevisiae and optimization of the pH conditions allowed for productive spontaneous rearrangement of salutaridinol-7-O-acetate and synthesis of thebaine from (R)-reticuline. Finally, we reconstituted a 7-gene pathway for the production of codeine and morphine from (R)-reticuline. Yeast cell feeding assays using (R)-reticuline, salutaridine or codeine as substrates showed that all enzymes were functionally co-expressed in yeast and that activity of salutaridine reductase and codeine-O-demethylase likely limit flux to morphine synthesis. The results of this study describe a significant advance for the synthesis of morphinans in S. cerevisiae and pave the way for their complete synthesis in recombinant microbes. PMID:25905794

  2. Constitutive Optimized Production of Streptokinase in Saccharomyces cerevisiae Utilizing Glyceraldehyde 3-Phosphate Dehydrogenase Promoter of Pichia pastoris

    OpenAIRE

    Ravi N. Vellanki; Ravichandra Potumarthi; Kiran K. Doddapaneni; Naveen Anubrolu; Mangamoori, Lakshmi N.

    2013-01-01

    A novel expression vector constructed from genes of Pichia pastoris was applied for heterologous gene expression in Saccharomyces cerevisiae. Recombinant streptokinase (SK) was synthesized by cloning the region encoding mature SK under the control of glyceraldehyde 3-phosphate dehydrogenase (GAP) promoter of Pichia pastoris in Saccharomyces cerevisiae. SK was intracellularly expressed constitutively, as evidenced by lyticase-nitroanilide and caseinolytic assays. The functional activity was co...

  3. Electrical stimulation of saccharomyces cerevisiae cultures Estimulação elétrica de células de Saccharomyces cerevisiae

    OpenAIRE

    Arau?jo, Ofelia Q. F.; Coelho, Maria Alice Z.; Margarit, Isabel C. P.; Vaz-junior, Carlos A.; Rocha-lea?o, Maria Helena M.

    2004-01-01

    Modulation of cell endogenous membrane potential by an external electrical field influences the structure and function of membrane compartments, proteins and lipid bi-layer. In this work, the effects of applied potential on Saccharomyces cerevisiae growth were characterized through simple yet conclusive experiments. Cell growth time profile and cell division were investigated as macroscopic response to the electrical stimulation. Control experiments were conducted under identical conditions e...

  4. Study on biosorption of uranium by alginate immobilized saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Saccharomyces cerevisiae has great capability of biosorption of uranium. The maxium uptake is 172.4 mg/g according to this study. To adapt to the application of the biomass in the field, the biosorption of uranium by cross-linked and alginate calcium immobilized Saccharomyces cerevisiae is studied. Results indicate the maxium uptake is 185.2 mg/g by formaldehyde cross-linked biomass, and it is 769.2 mg/g by alginate calcium immobilized biomass. (authors)

  5. Membrane Trafficking in the Yeast Saccharomyces cerevisiae Model

    Directory of Open Access Journals (Sweden)

    Serge Feyder

    2015-01-01

    Full Text Available The yeast Saccharomyces cerevisiae is one of the best characterized eukaryotic models. The secretory pathway was the first trafficking pathway clearly understood mainly thanks to the work done in the laboratory of Randy Schekman in the 1980s. They have isolated yeast sec mutants unable to secrete an extracellular enzyme and these SEC genes were identified as encoding key effectors of the secretory machinery. For this work, the 2013 Nobel Prize in Physiology and Medicine has been awarded to Randy Schekman; the prize is shared with James Rothman and Thomas Südhof. Here, we present the different trafficking pathways of yeast S. cerevisiae. At the Golgi apparatus newly synthesized proteins are sorted between those transported to the plasma membrane (PM, or the external medium, via the exocytosis or secretory pathway (SEC, and those targeted to the vacuole either through endosomes (vacuolar protein sorting or VPS pathway or directly (alkaline phosphatase or ALP pathway. Plasma membrane proteins can be internalized by endocytosis (END and transported to endosomes where they are sorted between those targeted for vacuolar degradation and those redirected to the Golgi (recycling or RCY pathway. Studies in yeast S. cerevisiae allowed the identification of most of the known effectors, protein complexes, and trafficking pathways in eukaryotic cells, and most of them are conserved among eukaryotes.

  6. PGM2 overexpression improves anaerobic galactose fermentation in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Hahn-Hägerdal Bärbel

    2010-05-01

    Full Text Available Abstract 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.

  7. Response of Saccharomyces cerevisiae to cadmium stress

    International Nuclear Information System (INIS)

    The intensification of industrial activity has been greatly contributing with the increase of heavy metals in the environment. Among these heavy metals, cadmium becomes a serious pervasive environmental pollutant. The cadmium is a heavy metal with no biological function, very toxic and carcinogenic at low concentrations. The toxicity of cadmium and several other metals can be mainly attributed to the multiplicity of coordination complexes and clusters that they can form. Some aspects of the cellular response to cadmium were extensively investigated in the yeast Saccharomyces cerevisiae. The primary site of interaction between many toxic metals and microbial cells is the plasma membrane. Plasma-membrane permeabilisation has been reported in a variety of microorganisms following cadmium exposure, and is considered one mechanism of cadmium toxicity in the yeast. In this work, using the yeast strain S. cerevisiae W303-WT, we have investigated the relationships between Cd uptake and release of cellular metal ions (K+ and Na+) using neutron activation technique. The neutron activation was an easy, rapid and suitable technique for doing these metal determinations on yeast cells; was observed the change in morphology of the strains during the process of Cd accumulation, these alterations were observed by Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) during incorporation of cadmium. (author)

  8. Senescence Mutants of Saccharomyces Cerevisiae with a Defect in Telomere Replication Identify Three Additional Est Genes

    OpenAIRE

    Lendvay, T. S.; Morris, D. K.; Sah, J.; Balasubramanian, B.; Lundblad, V.

    1996-01-01

    The primary determinant for telomere replication is the enzyme telomerase, responsible for elongating the G-rich strand of the telomere. The only component of this enzyme that has been identified in Saccharomyces cerevisiae is the TLC1 gene, encoding the telomerase RNA subunit. However, a yeast strain defective for the EST1 gene exhibits the same phenotypes (progressively shorter telomeres and a senescence phenotype) as a strain deleted for TLC1, suggesting that EST1 encodes either a componen...

  9. Cyclic AMP and Fluconazole Resistance in Saccharomyces cerevisiae

    OpenAIRE

    Kontoyiannis, D. P.; Rupp, Steffen

    2000-01-01

    Cyclic AMP (cAMP) is involved in the stress response in Saccharomyces cerevisiae. We show that cAMP is required for resistance to fluconazole in S. cerevisiae. In addition, activation of Ras2, a regulator of cAMP generation, results in some protection from fluconazole toxicity in a fashion independent of the efflux transporter Pdr5p.

  10. Molecular and enological characterization of a natural Saccharomyces uvarum and Saccharomyces cerevisiae hybrid.

    Science.gov (United States)

    Pérez-Torrado, Roberto; González, Sara Susana; Combina, Mariana; Barrio, Eladio; Querol, Amparo

    2015-07-01

    Saccharomyces cerevisiae plays a main role in the winemaking process, although other species, like Saccharomyces uvarum or Saccharomyces paradoxus, have been associated with must fermentations. It has been reported in recent years, that yeast hybrids of different Saccharomyces species might be responsible for wine productions. Although S. cerevisiae×Saccharomyces kudriavzevii hybrids have been well studied, very little attention has been paid to S. cerevisiae×S. uvarum hybrids. In this work we characterized the genomic composition of S6U, a widely used commercial S. cerevisiae×S. uvarum yeast hybrid isolated in wine fermentations containing one copy of the genome of each parental species, which suggests a relatively recent hybridization event. We also studied its performance under diverse enological conditions. The results show enhanced performance under low temperature enological conditions, increased glycerol production, lower acetic acid production and increased production of interesting aroma compounds. We also examined the transcriptomic response of the S6U hybrid strain compared with the reference species under enological conditions. The results show that although the hybrid strain transcriptome is more similar to S. uvarum than to S. cerevisiae, it presents specifically regulated genes involved in stress response, lipids and amino acid metabolism. The enological performance and aroma profile of this S. cerevisiae×S. uvarum hybrid makes it a good candidate for participating in winemaking, especially at low temperatures. PMID:25867085

  11. Redox balancing in recombinant strains of Saccharomyces cerevisiae

    Energy Technology Data Exchange (ETDEWEB)

    Anderlund, M.

    1998-09-01

    In metabolically engineered Saccharomyces cerevisiae expressing Pichia stipitis XYL1 and XYL2 genes, encoding xylose reductase (XR) and xylitol dehydrogenase (XDH), respectively, xylitol is excreted as the major product during anaerobic xylose fermentation and only low yields of ethanol are produced. This has been interpreted as a result of the dual cofactor dependence of XR and the exclusive use of NAD{sup +} by XDH. The excretion of xylitol was completely stopped and the formation of glycerol and acetic acid were reduced in xylose utilising S. cerevisiae strains cultivated in oxygen-limited conditions by expressing lower levels of XR than of XDH. The expression level of XYL1 and XYL2 were controlled by changing the promoters and transcription directions of the genes. A new functional metabolic pathway was established when Thermus thermophilus xylA gene was expressed in S. cerevisiae. The recombinant strain was able to ferment xylose to ethanol when cultivated on a minimal medium containing xylose as only carbon source. In order to create a channeled metabolic transfer in the two first steps of the xylose metabolism, XYL1 and XYL2 were fused in-frame and expressed in S. cerevisiae. When the fusion protein, containing a linker of three amino acids, was co expressed together with native XR and XDH monomers, enzyme complexes consisting of chimeric and native subunits were formed. The total activity of these complexes exhibited 10 and 9 times higher XR and XDH activity, respectively, than the original conjugates, consisting of only chimeric subunits. This strain produced less xylitol and the xylitol yield was lower than with strains only expressing native XR and XDH monomers. In addition, more ethanol and less acetic acid were formed. A new gene encoding the cytoplasmic transhydrogenase from Azotobacter vinelandii was cloned. The enzyme showed high similarity to the family of pyridine nucleotide-disulphide oxidoreductase. To analyse the physiological effect of transhydrogenation between the two coenzyme systems NADP(H) and NAD(H) during anaerobic growth, S. cerevisiae was transformed with a plasma membrane bound AB-transhydrogenase from E. coli and with a cytoplasmic BB-transhydrogenase from A. vinelandii. Expression of both types changed the intracellular nucleotide levels. The NADPH/NADP{sup +} ratio was reduced while the NADH/NAD{sup +} ratio was almost constant. An increased formation of 2-oxoglutarate, glycerol and acetate was observed during anaerobic glucose fermentation 206 refs, 8 figs, 3 tabs

  12. Directed Evolution towards Increased Isoprenoid Production in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Carlsen, Simon; Nielsen, Michael Lynge

    Due to declining drug discovery rates from organic synthetic libraries, pharmaceutical companies are turning their attention towards secondary metabolites. Isoprenoids, also known as terpenoids, constitute the largest known group of secondary metabolites isolated to date, encompassing more than 55,000 different compounds including several blockbuster drugs such as paclitaxel and artemisinin. All molecules within this group are biosynthesized from the same precursor called isopentenyl pyrophosphate (IPP), which is repeatedly polymerized and diversified giving rise to enormous chemical and structural diversity. The most common way of producing these compounds is by organic synthesis. Organic synthesis does however have several disadvantages for production of secondary metabolites such as low yields due to the complex structures, which makes this way of production economically unfeasible. Microbial production can easily be scaled to meet current demands and it is also an environmental benign production method compared to organic synthesis. Thus it would be attractive to engineer a microorganism to produce high amounts of IPP and other immediate prenyl precursors such as geranyl pyrophosphate, farnesyl pyrophosphate and geranylgeranyl pyrophosphate for large-scale microbial production of terpenoids. Saccharomyces cerevisiae was chosen as production platform due to its widespread use in industrial production and the waste number of molecular biology tools which is available for its manipulation. The effort for discovering new genetic perturbations, which would results in and increased production of isoprenoids by S. cerevisiae has been very limited. This project is focus on creating diversity within a lycopene producing S. cerevisiae strain by construction of gDNA-, cDNA-, and transposon-libraries. The diversified population of S. cerevisiae clones will afterwards be screened using the isoprenoid molecule lycopene as a model compound, hereby enabling the isolation of phenotypes producing higher amounts of isoprenoid. The property making lycopene ideal for screening is its system of 11 conjugated double bonds, which absorbs light within the visible range resulting in the red color of lycopene. This feature is the cause for the orange/red phenotype of S. cerevisiae strains transformed with the genes encoding lycopene and enables visual screening of yeast colonies, by searching for colonies with more intense red colony coloration which is the result of higher amount of lycopene is being produced and hence high amount of isoprenoid precursor being available. This will elucidate novel genetic targets for increasing isoprenoid production in S. cerevisiae

  13. Acetylation dynamics and stoichiometry in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Weinert, Brian Tate; Iesmantavicius, Vytautas

    2014-01-01

    Lysine acetylation is a frequently occurring posttranslational modification; however, little is known about the origin and regulation of most sites. Here we used quantitative mass spectrometry to analyze acetylation dynamics and stoichiometry in Saccharomyces cerevisiae. We found that acetylation accumulated in growth-arrested cells in a manner that depended on acetyl-CoA generation in distinct subcellular compartments. Mitochondrial acetylation levels correlated with acetyl-CoA concentration in vivo and acetyl-CoA acetylated lysine residues nonenzymatically in vitro. We developed a method to estimate acetylation stoichiometry and found that the vast majority of mitochondrial and cytoplasmic acetylation had a very low stoichiometry. However, mitochondrial acetylation occurred at a significantly higher basal level than cytoplasmic acetylation, consistent with the distinct acetylation dynamics and higher acetyl-CoA concentration in mitochondria. High stoichiometry acetylation occurred mostly on histones, proteins present in histone acetyltransferase and deacetylase complexes, and on transcription factors. These data show that a majority of acetylation occurs at very low levels in exponentially growing yeast and is uniformly affected by exposure to acetyl-CoA.

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

    OpenAIRE

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

    2007-01-01

    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 the two should be classified as conspecific. This raises the question of the distinguishing molecular...

  15. Switching the mode of sucrose utilization by Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Miletti Luiz C

    2008-02-01

    Full Text Available Abstract Background Overflow metabolism is an undesirable characteristic of aerobic cultures of Saccharomyces cerevisiae during biomass-directed processes. It results from elevated sugar consumption rates that cause a high substrate conversion to ethanol and other bi-products, severely affecting cell physiology, bioprocess performance, and biomass yields. Fed-batch culture, where sucrose consumption rates are controlled by the external addition of sugar aiming at its low concentrations in the fermentor, is the classical bioprocessing alternative to prevent sugar fermentation by yeasts. However, fed-batch fermentations present drawbacks that could be overcome by simpler batch cultures at relatively high (e.g. 20 g/L initial sugar concentrations. In this study, a S. cerevisiae strain lacking invertase activity was engineered to transport sucrose into the cells through a low-affinity and low-capacity sucrose-H+ symport activity, and the growth kinetics and biomass yields on sucrose analyzed using simple batch cultures. Results We have deleted from the genome of a S. cerevisiae strain lacking invertase the high-affinity sucrose-H+ symporter encoded by the AGT1 gene. This strain could still grow efficiently on sucrose due to a low-affinity and low-capacity sucrose-H+ symport activity mediated by the MALx1 maltose permeases, and its further intracellular hydrolysis by cytoplasmic maltases. Although sucrose consumption by this engineered yeast strain was slower than with the parental yeast strain, the cells grew efficiently on sucrose due to an increased respiration of the carbon source. Consequently, this engineered yeast strain produced less ethanol and 1.5 to 2 times more biomass when cultivated in simple batch mode using 20 g/L sucrose as the carbon source. Conclusion Higher cell densities during batch cultures on 20 g/L sucrose were achieved by using a S. cerevisiae strain engineered in the sucrose uptake system. Such result was accomplished by effectively reducing sucrose uptake by the yeast cells, avoiding overflow metabolism, with the concomitant reduction in ethanol production. The use of this modified yeast strain in simpler batch culture mode can be a viable option to more complicated traditional sucrose-limited fed-batch cultures for biomass-directed processes of S. cerevisiae.

  16. Accumulation of gold using Baker's yeast, Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Authors have reported preconcentration of 152Eu, a long-lived fission product, by yeast cells, Saccharomyces cerevisiae. Gold being a precious metal is used in electroplating, hydrogenation catalyst, etc. Heterogeneous composition of samples and low concentration offers renewed interest in its selective extraction of gold using various extractants. Gold can be recovered from different solutions using various chemical reagents like amines, organophosphorus compounds, and extractants containing sulphur as donor atom, etc. In the present work, two different strains of baker's yeast, Saccharomyces cerevisiae have been used to study the preconcentration of gold at various experimental conditions

  17. Structure of Ynk1 from the yeast Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    The crystal structure of Ynk1, an NDPK from the yeast Saccharomyces cerevisiae, has been solved at 3.1 Å resolution. Nucleoside diphosphate kinase (NDPK) catalyzes the transfer of the ?-phosphate from nucleoside triphosphates to nucleoside diphosphates. In addition to biochemical studies, a number of crystal structures of NDPK from various organisms, including both native proteins and complexes with nucleotides or nucleotide analogues, have been determined. Here, the crystal structure of Ynk1, an NDPK from the yeast Saccharomyces cerevisiae, has been solved at 3.1 Å resolution. Structural analysis strongly supports the oligomerization state of this protein being hexameric rather than tetrameric

  18. Regulation of Gluconeogenesis in Saccharomyces cerevisiae Is Mediated by Activator and Repressor Functions of Rds2?

    OpenAIRE

    Soontorngun, Nitnipa; Larochelle, Marc; Drouin, Simon; Robert, Franc?ois; Turcotte, Bernard

    2007-01-01

    In Saccharomyces cerevisiae, RDS2 encodes a zinc cluster transcription factor with unknown function. Here, we unravel a key function of Rds2 in gluconeogenesis using chromatin immunoprecipitation-chip technology. While we observed that Rds2 binds to only a few promoters in glucose-containing medium, it binds many additional genes when the medium is shifted to ethanol, a nonfermentable carbon source. Interestingly, many of these genes are involved in gluconeogenesis, the tricarboxylic acid cyc...

  19. Engineering and Analysis of a Saccharomyces cerevisiae Strain That Uses Formaldehyde as an Auxiliary Substrate?

    OpenAIRE

    Baerends, Richard J. S.; Hulster, Erik; Geertman, Jan-maarten A.; Daran, Jean-marc; Maris, Antonius J. A.; Veenhuis, Marten; Klei, Ida J.; Pronk, Jack T.

    2008-01-01

    We demonstrated that formaldehyde can be efficiently coutilized by an engineered Saccharomyces cerevisiae strain that expresses Hansenula polymorpha genes encoding formaldehyde dehydrogenase (FLD1) and formate dehydrogenase (FMD), in contrast to wild-type strains. Initial chemostat experiments showed that the engineered strain coutilized formaldehyde with glucose, but these mixed-substrate cultures failed to reach steady-state conditions and did not exhibit an increased biomass yield on gluco...

  20. Identification of a Cytosolically Directed NADH Dehydrogenase in Mitochondria of Saccharomyces cerevisiae

    OpenAIRE

    Small, W. Curtis; McAlister-Henn, Lee

    1998-01-01

    The reoxidation of NADH generated in reactions within the mitochondrial matrix of Saccharomyces cerevisiae is catalyzed by an NADH dehydrogenase designated Ndi1p (C. A. M. Marres, S. de Vries, and L. A. Grivell, Eur. J. Biochem. 195:857–862, 1991). Gene disruption analysis was used to examine possible metabolic functions of two proteins encoded by open reading frames having significant primary sequence similarity to Ndi1p. Disruption of the gene designated NDH1 results in a threefold reductio...

  1. Gateway Vectors for Efficient Artificial Gene Assembly In Vitro and Expression in Yeast Saccharomyces cerevisiae

    OpenAIRE

    Giuraniuc, Claudiu V.; Macpherson, Murray; Saka, Yasushi

    2013-01-01

    Construction of synthetic genetic networks requires the assembly of DNA fragments encoding functional biological parts in a defined order. Yet this may become a time-consuming procedure. To address this technical bottleneck, we have created a series of Gateway shuttle vectors and an integration vector, which facilitate the assembly of artificial genes and their expression in the budding yeast Saccharomyces cerevisiae. Our method enables the rapid construction of an artificia...

  2. Metabolic Engineering of Saccharomyces cerevisiae for Astaxanthin Production and Oxidative Stress Tolerance?

    OpenAIRE

    Ukibe, Ken; Hashida, Keisuke; Yoshida, Nobuyuki; Takagi, Hiroshi

    2009-01-01

    The red carotenoid astaxanthin possesses higher antioxidant activity than other carotenoids and has great commercial potential for use in the aquaculture, pharmaceutical, and food industries. In this study, we produced astaxanthin in the budding yeast Saccharomyces cerevisiae by introducing the genes involved in astaxanthin biosynthesis of carotenogenic microorganisms. In particular, expression of genes of the red yeast Xanthophyllomyces dendrorhous encoding phytoene desaturase (crtI product)...

  3. DNA polymerases delta and epsilon are required for chromosomal replication in Saccharomyces cerevisiae.

    OpenAIRE

    Budd, M. E.; Campbell, J. L.

    1993-01-01

    Three DNA polymerases, alpha, delta, and epsilon are required for viability in Saccharomyces cerevisiae. We have investigated whether DNA polymerases epsilon and delta are required for DNA replication. Two temperature-sensitive mutations in the POL2 gene, encoding DNA polymerase epsilon, have been identified by using the plasmid shuffle technique. Alkaline sucrose gradient analysis of DNA synthesis products in the mutant strains shows that no chromosomal-size DNA is formed after shift of an a...

  4. Genomewide Screen Reveals a Wide Regulatory Network for Di/Tripeptide Utilization in Saccharomyces cerevisiae

    OpenAIRE

    Cai, Houjian; Kauffman, Sarah; Naider, Fred; Becker, Jeffrey M.

    2006-01-01

    Small peptides of two to six residues serve as important sources of amino acids and nitrogen required for growth by a variety of organisms. In the yeast Saccharomyces cerevisiae, the membrane transport protein Ptr2p, encoded by PTR2, mediates the uptake of di/tripeptides. To identify genes involved in regulation of dipeptide utilization, we performed a systematic, functional examination of this process in a haploid, nonessential, single-gene deletion mutant library. We have identified 103 can...

  5. Metabolism of sulfur amino acids in Saccharomyces cerevisiae.

    OpenAIRE

    Thomas, D.; Surdin-Kerjan, Y

    1997-01-01

    Sulfur amino acid biosynthesis in Saccharomyces cerevisiae involves a large number of enzymes required for the de novo biosynthesis of methionine and cysteine and the recycling of organic sulfur metabolites. This review summarizes the details of these processes and analyzes the molecular data which have been acquired in this metabolic area. Sulfur biochemistry appears not to be unique through terrestrial life, and S. cerevisiae is one of the species of sulfate-assimilatory organisms possessin...

  6. Elevated evolutionary rates in the laboratory strain of Saccharomyces cerevisiae

    OpenAIRE

    Gu, Zhenglong; David, Lior; Petrov, Dmitri; Jones, Ted; Davis, Ronald W.; Lars M. Steinmetz

    2005-01-01

    By using the maximum likelihood method, we made a genome-wide comparison of the evolutionary rates in the lineages leading to the laboratory strain (S288c) and a wild strain (YJM789) of Saccharomyces cerevisiae and found that genes in the laboratory strain tend to evolve faster than in the wild strain. The pattern of elevated evolution suggests that relaxation of selection intensity is the dominant underlying reason, which is consistent with recurrent bottlenecks in the S. cerevisiae laborato...

  7. Research on biosorption of uranium by saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    The effects of pH and the granularity of S. cerevisiae on the biosorption capacity were examined in order to study the properties of the biosorption of uranium from effluent by Saccharomyces cerevisiae. The isotherm was drawn. From the isotherm, the equations of Langmuir and Freundlich were achieved. The results showed the highest biosorption capacity was obtained when the pH value was about 6 and the granularity was 0.15-0.13 mm

  8. The Saccharomyces cerevisiae Sgs1 helicase efficiently unwinds G-G paired DNAs.

    OpenAIRE

    Sun, H.; Bennett, R J; Maizels, N

    1999-01-01

    The Saccharomyces cerevisiae Sgs1p helicase localizes to the nucleolus and is required to maintain the integrity of the rDNA repeats. Sgs1p is a member of the RecQ DNA helicase family, which also includes Schizo-saccharomyces pombe Rqh1, and the human BLM and WRN genes. These genes encode proteins which are essential to maintenance of genomic integrity and which share a highly conserved helicase domain. Here we show that recombinant Sgs1p helicase efficiently unwinds guanine-guanine (G-G) pai...

  9. The Exocyst is a multiprotein complex required for exocytosis in Saccharomyces cerevisiae.

    OpenAIRE

    Terbush, D. R.; Maurice, T.; Roth, D.; Novick, P.

    1996-01-01

    In the yeast Saccharomyces cerevisiae, the products of at least 15 genes are involved specifically in vesicular transport from the Golgi apparatus to the plasma membrane. Previously, we have shown that three of these genes, SEC6, SEC8 and SEC15, encode components of a multisubunit complex which localizes to the tip of the bud, the predominant site of exocytosis in S. cerevisiae. Mutations in three more of these genes, SEC3, SEC5 and SEC10, were found to disrupt the subunit integrity of the Se...

  10. Divergence in wine characteristics produced by wild and domesticated strains of Saccharomyces cerevisiae

    OpenAIRE

    Hyma, Katie E.; Saerens, Sofie M; Kevin J. Verstrepen; Fay, Justin C

    2011-01-01

    The budding yeast Saccharomyces cerevisiae is the primary species used by wine makers to convert sugar into alcohol during wine fermentation. Saccharomyces cerevisiae is found in vineyards, but is also found in association with oak trees and other natural sources. Although wild strains of S. cerevisiae as well as other Saccharomyces species are also capable of wine fermentation, a genetically distinct group of S. cerevisiae strains is primarily used to produce wine, consistent with the idea t...

  11. CNS1 Encodes an Essential p60/Sti1 Homolog in Saccharomyces cerevisiae That Suppresses Cyclophilin 40 Mutations and Interacts with Hsp90

    OpenAIRE

    Dolinski, Kara J.; Cardenas, Maria E; Heitman, Joseph

    1998-01-01

    Cyclophilins are cis-trans-peptidyl-prolyl isomerases that bind to and are inhibited by the immunosuppressant cyclosporin A (CsA). The toxic effects of CsA are mediated by the 18-kDa cyclophilin A protein. A larger cyclophilin of 40 kDa, cyclophilin 40, is a component of Hsp90-steroid receptor complexes and contains two domains, an amino-terminal prolyl isomerase domain and a carboxy-terminal tetratricopeptide repeat (TPR) domain. There are two cyclophilin 40 homologs in the yeast Saccharomyc...

  12. Hydrogen peroxide removal with magnetically responsive Saccharomyces cerevisiae cells.

    Czech Academy of Sciences Publication Activity Database

    Šafa?ík, Ivo; Mad?rová, Zde?ka; Šafa?íková, Miroslava

    2008-01-01

    Ro?. 56, - (2008), s. 7925-7928. ISSN 0021-8561 R&D Projects: GA MPO 2A-1TP1/094; GA MŠk OC 157 Institutional research plan: CEZ:AV0Z60870520 Keywords : magnetic alginate beads * catalase * magnetic separation * Saccharomyces cerevisiae cells * hydrogen peroxide Subject RIV: GM - Food Processing Impact factor: 2.562, year: 2008

  13. Evidence against a photoprotective component of photoreactivation in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Photoreactivation-deficient (phr-) mutants of Saccharomyces cerevisiae were shown to lack in vitro DNA-photolyase activity. A phr- mutant was then compared with a phr+ strain for near-UV induced photoprotection from far-UV irradiation. Neither strain exhibited a photoprotective effect. (author)

  14. Kinetics of formation of induced mutants of Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    UV and ?-radiation mutagenic effect an various strains of Saccharomyces cerevisiae was studied by analyzing formation kinetics of induced mutants at the period of postirradiation incubation. Mechanisms of induced reverse formation was suggested. The presented analysis is considered to be differential taking account of more subtle aspects of induced mutagenesis. 8 refs.; 10 figs.; 3 tabs

  15. Interaction between Hanseniaspora uvarum and Saccharomyces cerevisiae during alcoholic fermentation.

    Science.gov (United States)

    Wang, Chunxiao; Mas, Albert; Esteve-Zarzoso, Braulio

    2015-08-01

    During wine fermentation, Saccharomyces clearly dominate over non-Saccharomyces wine yeasts, and several factors could be related to this dominance. However, the main factor causing the reduction of cultivable non-Saccharomyces populations has not yet been fully established. In the present study, various single and mixed fermentations were performed to evaluate some of the factors likely responsible for the interaction between Saccharomyces cerevisiae and Hanseniaspora uvarum. Alcoholic fermentation was performed in compartmented experimental set ups with ratios of 1:1 and 1:9 and the cultivable population of both species was followed. The cultivable H. uvarum population decreased sharply at late stages when S. cerevisiae was present in the other compartment, similarly to alcoholic fermentations in non-compartmented vessels. Thus, cell-to-cell contact did not seem to be the main cause for the lack of cultivability of H. uvarum. Other compounds related to fermentation performance (such as sugar and ethanol) and/or certain metabolites secreted by S. cerevisiae could be related to the sharp decrease in H. uvarum cultivability. When these factors were analyzed, it was confirmed that metabolites from S. cerevisiae induced lack of cultivability in H. uvarum, however ethanol and other possible compounds did not seem to induce this effect but played some role during the process. This study contributes to a new understanding of the lack of cultivability of H. uvarum populations during the late stages of wine fermentation. PMID:25956738

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

    Science.gov (United States)

    Deutch, Charles E.; Marshall, Pamela A.

    2008-01-01

    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…

  17. In vitro screening of probiotic properties of Saccharomyces cerevisiae var. boulardii and food-borne Saccharomyces cerevisiae strains

    DEFF Research Database (Denmark)

    van der Aa Kuhle, Alis; Skovgaard, Kerstin

    2005-01-01

    The probiotic potential of IS Saccharomyces cerevisiae strains used for production of foods or bevel-ages or isolated from such, and eight strains of Saccharomyces cerevisiae var. boulardii, was investigated. All strains included were able to withstand pH 2.5 and 0.3% Ox-all. Adhesion to the nontumorigenic porcine jejunal epithelial cell line (IPEC-J2) was investigated by incorporation of H-3-methionine into the yeast cells and use of liquid scintillation counting. Only few of the food-borne S. cerevisiae strains exhibited noteworthy adhesiveness with the strongest levels of adhesion (13.6-16.8%) recorded for two isolates from blue veined cheeses. Merely 25% of the S. cerevisiae var. boulardii strains displayed good adhesive properties (16.2-28.0%). The expression of the proinflammatory cytokine IL-1? decreased strikingly in IPEC-J2 cells exposed to a Shiga-like toxin 2e producing Escherichia coli strain when the cells were pre- and coincubated with S. cerevisiae var. boulardii even though this yeast strain was low adhesive (5.4%), suggesting that adhesion is not a mandatory prerequisite for such a probiotic effect. A strain of S. cerevisiae isolated from West African sorghum beer exerted similar effects hence indicating that food-borne strains of S. cerevisiae may possess probiotic properties in spite of low adhesiveness. © 2004 Elsevier B.V. All rights reserved.

  18. Functional studies of aldo-keto reductases in Saccharomyces cerevisiae*

    Science.gov (United States)

    Chang, Qing; Griest, Terry A.; Harter, Theresa M.; Petrash, J. Mark

    2007-01-01

    SUMMARY We utilized the budding yeast Saccharomyces cerevisiae as a model to systematically explore physiological roles for yeast and mammalian aldo-keto reductases. Six open reading frames encoding putative aldo-keto reductases were identified when the yeast genome was queried against the sequence for human aldose reductase, the prototypical mammalian aldo-keto reductase. Recombinant proteins produced from five of these yeast open reading frames demonstrated NADPH-dependent reductase activity with a variety of aldehyde and ketone substrates. A triple aldo-keto reductase null mutant strain demonstrated a glucose-dependent heat shock phenotype which could be rescued by ectopic expression of human aldose reductase. Catalytically-inactive mutants of human or yeast aldo-keto reductases failed to effect a rescue of the heat shock phenotype, suggesting that the phenotype results from either an accumulation of one or more unmetabolized aldo-keto reductase substrates or a synthetic deficiency of aldo-keto reductase products generated in response to heat shock stress. These results suggest that multiple aldo-keto reductases fulfill functionally redundant roles in the stress response in yeast. PMID:17140678

  19. Functional studies of aldo-keto reductases in Saccharomyces cerevisiae.

    Science.gov (United States)

    Chang, Qing; Griest, Terry A; Harter, Theresa M; Petrash, J Mark

    2007-03-01

    We utilized the budding yeast Saccharomyces cerevisiae as a model to systematically explore physiological roles for yeast and mammalian aldo-keto reductases. Six open reading frames encoding putative aldo-keto reductases were identified when the yeast genome was queried against the sequence for human aldose reductase, the prototypical mammalian aldo-keto reductase. Recombinant proteins produced from five of these yeast open reading frames demonstrated NADPH-dependent reductase activity with a variety of aldehyde and ketone substrates. A triple aldo-keto reductase null mutant strain demonstrated a glucose-dependent heat shock phenotype which could be rescued by ectopic expression of human aldose reductase. Catalytically-inactive mutants of human or yeast aldo-keto reductases failed to effect a rescue of the heat shock phenotype, suggesting that the phenotype results from either an accumulation of one or more unmetabolized aldo-keto reductase substrates or a synthetic deficiency of aldo-keto reductase products generated in response to heat shock stress. These results suggest that multiple aldo-keto reductases fulfill functionally redundant roles in the stress response in yeast. PMID:17140678

  20. Biosorption of 241Am by immobilized Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Americium-241 is one of the most serious radioactive contaminating nuclides due to its high toxicity and long half-life. The encouraging biosorption of 241Am from aqueous solutions by free Saccharomyces cerevisiae (S. cerevisiae) has been observed in our previous experiments. 241Am biosorption by immobilized S. cerevisiae and the effect of the various experimental conditions on the adsorption were investigated. The results indicated that the 241Am biosorption by immobilized S. cerevisiae is still very efficient, and immobilized S. cerevisiae can be used repeatedly or continuously. The biosorption equilibrium was achieved within 2 hours, and more than 92% of 241Am was removed by immobilized S. cerevisiae in the pH 1-4 range. No significant differences in 241Am biosorption were observed at 15-45 deg C. The immobilized S. cerevisiae, even after used repeatedly for 6 times, still could adsorb more than 90% of 241Am in solutions of 1.08 MBq/l (8.5 ?g/l). At this moment, the total adsorption capacity for 241Am was more than 63.3 KBq/g globe (0.5 ?g/g), but has not reached saturation yet. The 241Am left in solutions with initial concentration of 1.08 MBq/l (8.5 ?g/l) was noted as low as ?10 Bq/l (?8.0 x 10-5 ?g/l) after adsorption by the immobilized S. cerevisiae for 3 times. (author)

  1. Potential immobilized Saccharomyces cerevisiae as heavy metal removal

    Science.gov (United States)

    Raffar, Nur Izzati Abdul; Rahman, Nadhratul Nur Ain Abdul; Alrozi, Rasyidah; Senusi, Faraziehan; Chang, Siu Hua

    2015-05-01

    Biosorption of copper ion using treated and untreated immobilized Saccharomyces cerevisiae from aqueous solution was investigate in this study. S.cerevisiae has been choosing as biosorbent due to low cost, easy and continuously available from various industries. In this study, the ability of treated and untreated immobilized S.cerevisiae in removing copper ion influence by the effect of pH solution, and initial concentration of copper ion with contact time. Besides, adsorption isotherm and kinetic model also studied. The result indicated that the copper ion uptake on treated and untreated immobilized S.cerevisiae was increased with increasing of contact time and initial concentration of copper ion. The optimum pH for copper ion uptake on untreated and treated immobilized S.cerevisiae at 4 and 6. From the data obtained of copper ion uptake, the adsorption isotherm was fitted well by Freundlich model for treated immobilized S.cerevisiae and Langmuir model for untreated immobilized S.cerevisiae according to high correlation coefficient. Meanwhile, the pseudo second order was described as suitable model present according to high correlation coefficient. Since the application of biosorption process has been received more attention from numerous researchers as a potential process to be applied in the industry, future study will be conducted to investigate the potential of immobilized S.cerevisiae in continuous process.

  2. Electrical stimulation of saccharomyces cerevisiae cultures / Estimulação elétrica de células de Saccharomyces cerevisiae

    Scientific Electronic Library Online (English)

    Ofelia Q.F., Araújo; Maria Alice Z., Coelho; Isabel C.P., Margarit; Carlos A., Vaz-Junior; Maria Helena M., Rocha-Leão.

    2004-06-01

    Full Text Available Modulação do potencial de membrana celular endógeno por um campo elétrico externo influencia a estrutura e função dos compartimentos da membrana, de suas proteínas e da bi-camada lipídica. Neste trabalho, os efeitos da aplicação de potencial no crescimento de Saccharomyces cerevisiae foram caracteri [...] zados por experimentos simples, mas conclusivos. O perfil temporal de crescimento celular e a divisão celular foram investigados como respostas macroscópicas ao estímulo elétrico. Experimentos controle foram conduzidos em condições idênticas, exceto pela ausência de potencial aplicado. Através de análise comparativa, verificou-se que o estímulo elétrico alterou o ciclo celular como foi possível observar através da medida da dispersão de tamanho celular de cada população, sugerindo um possível sincronismo na divisão celular. Análise do espectro de potência foi empregada para sustentar o aumento no sincronismo, e uma modelagem matemática foi conduzida para determinar mudanças na cinética de crescimento celular. Parâmetros cinéticos do modelo tipo Monod para crescimento foram determinados por regressão não-linear. A constante de afinidade (a saber, K S) apresentou uma dependência com o potencial aplicado, sugerindo mudanças no transporte através da membrana celular. Verificou-se, também, que o estresse promovido eletroquimicamente inibiu o crescimento e induziu mudanças na viabilidade celular. Abstract in english Modulation of cell endogenous membrane potential by an external electrical field influences the structure and function of membrane compartments, proteins and lipid bi-layer. In this work, the effects of applied potential on Saccharomyces cerevisiae growth were characterized through simple yet conclu [...] sive experiments. Cell growth time profile and cell division were investigated as macroscopic response to the electrical stimulation. Control experiments were conducted under identical conditions except for the absence of applied potential. Through comparative analysis, electrical stimulation was verified to alter cell cycle as smaller sized population was observed, suggesting that a synchrony in cell division was promoted. Power spectral analysis was employed to sustain synchrony enhancement, and mathematical modeling was conducted for determining kinetic growth changes. Monod type kinetic parameters for growth were determined by non-linear regression. The affinity constant (namely kS) presented a dependence on applied potential suggesting changes on transport across cell membrane. Electrochemically promoted stress was also verified to inhibit growth as well as to induce changes on cell viability.

  3. Evolutionary engineering of Saccharomyces cerevisiae for efficient aerobic xylose consumption.

    DEFF Research Database (Denmark)

    Scalcinati, Gionata; Otero, José Manuel

    2012-01-01

    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 utilizationrates. 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.

  4. Evidence for Domesticated and Wild Populations of Saccharomyces cerevisiae.

    Directory of Open Access Journals (Sweden)

    2005-07-01

    Full Text Available 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 isolates of S. cerevisiae from other sources simply represent migrants from these fermentations. We have surveyed DNA sequence diversity at five loci in 81 strains of S. cerevisiae that were isolated from a variety of human and natural fermentations as well as sources unrelated to alcoholic beverage production, such as tree exudates and immunocompromised patients. Diversity within vineyard strains and within saké strains is low, consistent with their status as domesticated stocks. The oldest lineages and the majority of variation are found in strains from sources unrelated to wine production. We propose a model whereby two specialized breeds of S. cerevisiae have been created, one for the production of grape wine and one for the production of saké wine. We estimate that these two breeds have remained isolated from one another for thousands of years, consistent with the earliest archeological evidence for winemaking. We conclude that although there are clearly strains of S. cerevisiae specialized for the production of alcoholic beverages, these have been derived from natural populations unassociated with alcoholic beverage production, rather than the opposite.

  5. A Member of the Sugar Transporter Family, Stl1p Is the Glycerol/H+ Symporter in Saccharomyces cerevisiae

    OpenAIRE

    Ferreira, Ce?lia; Voorst, Frank; Martins, Anto?nio; Neves, Luisa; Oliveira, Rui; Kielland-brandt, Morten C.; Lucas, Ca?ndida; Brandt, Anders

    2005-01-01

    Glycerol and other polyols are used as osmoprotectants by many organisms. Several yeasts and other fungi can take up glycerol by proton symport. To identify genes involved in active glycerol uptake in Saccharomyces cerevisiae we screened a deletion mutant collection comprising 321 genes encoding proteins with 6 or more predicted transmembrane domains for impaired growth on glycerol medium. Deletion of STL1, which encodes a member of the sugar transporter family, eliminates active glycerol tra...

  6. Unlinked Noncomplementation: Isolation of New Conditional-Lethal Mutations in Each of the Tubulin Genes of Saccharomyces Cerevisiae

    OpenAIRE

    Stearns, T.; Botstein, D.

    1988-01-01

    Mutations in genes of Saccharomyces cerevisiae that code for proteins that interact with ?-tubulin were sought by screening for unlinked mutations that fail to complement mutations in the single ?-tubulin-encoding gene (TUB2). Among the first three noncomplementing mutations examined, two are linked to TUB2 while one is unlinked. The unlinked mutation was shown to be a conditional-lethal allele of the major ?-tubulin-encoding gene (TUB1) and represents the first such mutation in that gene....

  7. Cloning and characterization of Kluyveromyces lactis SEC14, a gene whose product stimulates Golgi secretory function in Saccharomyces cerevisiae.

    OpenAIRE

    Salama, S. R.; Cleves, A. E.; Malehorn, D. E.; Whitters, E. A.; Bankaitis, V. A.

    1990-01-01

    The Saccharomyces cerevisiae SEC14 gene encodes a cytosolic factor that is required for secretory protein movement from the Golgi complex. That some conservation of SEC14p function may exist was initially suggested by experiments that revealed immunoreactive polypeptides in cell extracts of the divergent yeasts Kluyveromyces lactis and Schizosaccharomyces pombe. We have cloned and characterized the K. lactis SEC14 gene (SEC14KL). Immunoprecipitation experiments indicated that SEC14KL encoded ...

  8. Influence of organic acids and organochlorinated insecticides on metabolism of Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Pejin Dušanka J.

    2005-01-01

    Full Text Available Saccharomyces cerevisiae is exposed to different stress factors during the production: osmotic, temperature, oxidative. The response to these stresses is the adaptive mechanism of cells. The raw materials Saccharomyces cerevisiae is produced from, contain metabolism products of present microorganisms and protective agents used during the growth of sugar beet for example the influence of acetic and butyric acid and organochlorinated insecticides, lindan and heptachlor, on the metabolism of Saccharomyces cerevisiae was investigated and presented in this work. The mentioned compounds affect negatively the specific growth rate, yield, content of proteins, phosphorus, total ribonucleic acids. These compounds influence the increase of trechalose and glycogen content in the Saccharomyces cerevisiae cells.

  9. Intracellular ethanol accumulation in Saccharomyces cerevisiae during fermentation.

    OpenAIRE

    D'Amore, T; Panchal, C J; Stewart, G G

    1988-01-01

    An intracellular accumulation of ethanol in Saccharomyces cerevisiae was observed during the early stages of fermentation (3 h). However, after 12 h of fermentation, the intracellular and extracellular ethanol concentrations were similar. Increasing the osmotic pressure of the medium caused an increase in the ratio of intracellular to extracellular ethanol concentrations at 3 h of fermentation. As in the previous case, the intracellular and extracellular ethanol concentrations were similar af...

  10. Biogeographical characterization of Saccharomyces cerevisiae wine yeast by molecular methods

    OpenAIRE

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

    2013-01-01

    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 pract...

  11. Purification of Arp2/3 complex from Saccharomyces cerevisiae

    OpenAIRE

    Doolittle, Lynda K; Rosen, Michael K.; Padrick, Shae B.

    2013-01-01

    Much of cellular control over actin dynamics comes through regulation of actin filament initiation. At the molecular level, this is accomplished through a collection of cellular protein machines, called actin nucleation factors, which position actin monomers to initiate a new actin filament. The Arp2/3 complex is a principal actin nucleation factor used throughout the eukaryotic family tree. The budding yeast Saccharomyces cerevisiae has proven to be not only an excellent genetic platform for...

  12. Purification and analysis of checkpoint protein complexes from Saccharomyces cerevisiae.

    OpenAIRE

    Green, CM; Lowndes, NF

    2004-01-01

    The DNA damage-dependent checkpoint of Saccharomyces cerevisiae is a paradigm for eukaryotic checkpoint pathways that regulate cell cycle progression in the presence of insults to the genetic material. In order to better understand this pathway, we undertook a biochemical study of the proteins implicated in its functioning. Analysis of the hydrodynamic properties of a protein in a crude mixture can give insights into possible tertiary organization such as participation in high-molecular-mass ...

  13. Magnetically altered ethanol fermentation capacity of Saccharomyces cerevisiae

    OpenAIRE

    Galonja-Corghill Tamara; Kostadinovi? Ljiljana M.; Bojat Nenad C.

    2009-01-01

    We studied the effect of static magnetic fields on ethanol production by yeast Saccharomyces cerevisiae 424A (LNH-ST) using sugar cane molasses during the fermentation in an enclosed bioreactor. Two static NdFeB magnets were attached to a cylindrical tube reactor with their opposite poles (north to south), creating 150 mT magnetic field inside the reactor. Comparable differences emerged between the results of these two experimental conditions. We found ethanol productivity to be 15% higher in...

  14. Genetic and biochemical characterization of clathrin-deficient Saccharomyces cerevisiae.

    OpenAIRE

    Payne, G. S.; Hasson, T B; Hasson, M S; Schekman, R.

    1987-01-01

    Clathrin is important but not essential for yeast cell growth and protein secretion. Diploid Saccharomyces cerevisiae cells heterozygous for a clathrin heavy-chain gene (CHC1) disruption give rise to viable, slow-growing, clathrin heavy-chain-deficient meiotic progeny (G. Payne and R. Schekman, Science 230:1009-1014, 1985). The possibility that extragenic suppressors account for growth of clathrin-deficient cells was examined by deletion of CHC1 from haploid cell genomes by single-step gene t...

  15. Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems

    OpenAIRE

    DiCarlo, James; Norville, Julie; Mali, Prashant; Rios Villanueva, Xavier; Aach, John Dennis; Church, George McDonald

    2013-01-01

    Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated (Cas) systems in bacteria and archaea use RNA-guided nuclease activity to provide adaptive immunity against invading foreign nucleic acids. Here, we report the use of type II bacterial CRISPR-Cas system in Saccharomyces cerevisiae for genome engineering. The CRISPR-Cas components, Cas9 gene and a designer genome targeting CRISPR guide RNA (gRNA), show robust and specific RNA-guided endonuclease activity a...

  16. Membrane-associated phosphatidylinositol kinase from Saccharomyces cerevisiae.

    OpenAIRE

    McKenzie, M A; Carman, G M

    1983-01-01

    Membrane-associated phosphatidylinositol kinase (ATP:phosphatidylinositol 4-phosphotransferase, EC 2.7.1.67) was partially purified 93-fold from Saccharomyces cerevisiae. Activity was dependent on magnesium ions (10 mM) and the optimum pH was 8.5. The apparent Km values for ATP and phosphatidylinositol were 0.21 mM and 71 microM, respectively. Activity was stimulated by sodium cholate and inhibited by sodium, potassium, lithium, and fluoride ions.

  17. Adenine deaminase and adenine utilization in Saccharomyces cerevisiae.

    OpenAIRE

    Deeley, M. C.

    1992-01-01

    Compared with other purine salvage and nitrogen catabolism enzymatic activities, adenine deaminase (adenine aminohydrolase [AAH]; EC 3.5.4.2) activity in Saccharomyces cerevisiae is uniquely regulated. AAH specific activity is not induced by adenine and is reduced sevenfold when cells are cultivated in medium containing proline in place of ammonium as the sole nitrogen source. Exogenous adenine enters metabolic pathways primarily via the function of either AAH or adenine phosphoribosyltransfe...

  18. Coordinate regulation of phospholipid biosynthesis by serine in Saccharomyces cerevisiae.

    OpenAIRE

    Homann, M J; Bailis, A. M.; Henry, S. A.; Carman, G. M.

    1987-01-01

    The addition of L-serine to inositol-containing growth medium repressed membrane-associated CDPdiacylglycerol synthase (CTP:phosphatidate cytidylyltransferase, EC 2.7.7.41) and phosphatidylserine synthase (CDPdiacylglycerol:L-serine O-phosphatidyltransferase, EC 2.7.8.8) activities and subunit levels in wild-type Saccharomyces cerevisiae. Enzyme activities and subunit levels were not repressed when inositol was absent from the growth medium. The addition of L-serine to the growth medium did n...

  19. Regulation of phosphatidate phosphatase activity by inositol in Saccharomyces cerevisiae.

    OpenAIRE

    Morlock, K R; Lin, Y P; Carman, G. M.

    1988-01-01

    Regulation of phosphatidate phosphatase (EC 3.1.34) activity was examined in Saccharomyces cerevisiae cells supplemented with phospholipid precursors. Addition of inositol to the growth medium of wild-type cells resulted in a twofold increase in phosphatidate phosphatase activity. The increase in phosphatidate phosphatase activity was not due to soluble effector molecules, and inositol did not have a direct effect on enzyme activity. The phosphatidate phosphatase activity associated with the ...

  20. Size control models of Saccharomyces cerevisiae cell proliferation.

    OpenAIRE

    Wheals, A. E.

    1982-01-01

    By using time-lapse photomicroscopy, the individual cycle times and sizes at bud emergence were measured for a population of saccharomyces cerevisiae cells growing exponentially under balanced growth conditions in a specially constructed filming slide. There was extensive variability in both parameters for daughter and parent cells. The data on 162 pairs of siblings were analyzed for agreement with the predictions of the transition probability hypothesis and the critical-size hypothesis of ye...

  1. Regulation of Phospholipid Synthesis in the Yeast Saccharomyces cerevisiae

    OpenAIRE

    CARMAN, George M.; Han, Gil-Soo

    2011-01-01

    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. P...

  2. Spontaneous loss of heterozygosity in diploid Saccharomyces cerevisiae cells.

    OpenAIRE

    Hiraoka, M; Watanabe, K.; Umezu, K; Maki, H.

    2000-01-01

    To obtain a broad perspective of the events leading to spontaneous loss of heterozygosity (LOH), we have characterized the genetic alterations that functionally inactivated the URA3 marker hemizygously or heterozygously situated either on chromosome III or chromosome V in diploid Saccharomyces cerevisiae cells. Analysis of chromosome structure in a large number of LOH clones by pulsed-field gel electrophoresis and PCR showed that chromosome loss, allelic recombination, and chromosome aberrati...

  3. Analysis of feedback-resistant anthranilate synthases from Saccharomyces cerevisiae.

    OpenAIRE

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

    1993-01-01

    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(...

  4. Clustering of the Genes for Allantoin Degradation in Saccharomyces cerevisiae

    Science.gov (United States)

    Lawther, Robert P.; Riemer, Elaine; Chojnacki, Bonnie; Cooper, Terrance G.

    1974-01-01

    We have shown that allantoin degradation in Saccharomyces cerevisiae proceeds exclusively through the intermediate formation of allantoic acid, urea, and allophanic acid. The number of reactions between allantoic acid and urea, however, remains obscure owing to our inability to isolate a mutant defective in ureidoglycolate hydrolase. Structural genes for the enzymes, allantoinase (dal1) and allantoicase (dal2) are located on chromosome IX promixal to the centromere in the order dal1-dal2-lysl. PMID:4604238

  5. Reversal of PCNA Ubiquitylation by Ubp10 in Saccharomyces cerevisiae

    OpenAIRE

    Gallego-sa?nchez, Alfonso; Andre?s, Sonia; Conde, Francisco; San Segundo, Pedro; Bueno, Avelino

    2012-01-01

    Regulation of PCNA ubiquitylation plays a key role in the tolerance to DNA damage in eukaryotes. Although the evolutionary conserved mechanism of PCNA ubiquitylation is well understood, the deubiquitylation of ubPCNA remains poorly characterized. Here, we show that the histone H2B K123 ubiquitin protease Ubp10 also deubiquitylates ubPCNA in Saccharomyces cerevisiae. Our results sustain that Ubp10-dependent deubiquitylation of the sliding clamp PCNA normally takes place during S phase, likely ...

  6. Heterologous Production of Dihomo-?-Linolenic Acid in Saccharomyces cerevisiae?

    OpenAIRE

    Yazawa, Hisashi; Iwahashi, Hitoshi; Kamisaka, Yasushi; Kimura, Kazuyoshi; Aki, Tsunehiro; Ono, Kazuhisa; Uemura, Hiroshi

    2007-01-01

    To make dihomo-?-linolenic acid (DGLA) (20:3n-6) in Saccharomyces cerevisiae, we introduced Kluyveromyces lactis ?12 fatty acid desaturase, rat ?6 fatty acid desaturase, and rat elongase genes. Because Fad2p is able to convert the endogenous oleic acid to linoleic acid, this allowed DGLA biosynthesis without the need to supply exogenous fatty acids on the media. Medium composition, cultivation temperature, and incubation time were examined to improve the yield of DGLA. Fatty acid content w...

  7. Investigation of nutrient sensing in the yeast Saccharomyces cerevisiae

    OpenAIRE

    Eckert-boulet, Nadine; Hobley, Timothy John

    2006-01-01

    Gæren Saccharomyces cerevisiae har udviklet komplekse regulatoriske systemer til at kontrollere ekspression af de proteiner, der importerer næringsstoffer, således at disse kun bliver produceret, når der er brug for dem. Dette er tilfældet for hexose-transportører samt aminosyre-transportører (disse bliver også kaldt amino acid permeases (AAPs)). Deres ekspression induceres på det transkriptionelle niveau efter at ekstracellulære næringsstoffer, henholdsvis glukose og aminosyrer, b...

  8. Shuttle mutagenesis: a method of transposon mutagenesis for Saccharomyces cerevisiae.

    OpenAIRE

    Seifert, H S; Chen, E Y; So, M.; Heffron, F

    1986-01-01

    We have extended the method of transposon mutagenesis to the eukaryote, Saccharomyces cerevisiae. A bacterial transposon containing a selectable yeast gene can be transposed into a cloned fragment of yeast DNA in Escherichia coli, and the transposon insertion can be returned to the yeast genome by homologous recombination. Initially, the cloned yeast DNA fragment to be mutagenized was transformed into an E. coli strain containing an F factor derivative carrying the transposable element. The c...

  9. Coordinate regulation of phosphatidylserine decarboxylase in Saccharomyces cerevisiae.

    OpenAIRE

    Lamping, E.; Kohlwein, S. D.; Henry, S. A.; Paltauf, F.

    1991-01-01

    Regulation of the activity of the mitochondrial enzyme phosphatidylserine decarboxylase (PSD) was measured in vitro by using membrane preparations from wild-type and mutant strains of Saccharomyces cerevisiae. PSD specific activity was not affected by carbon source, and on all carbon sources, the highest specific activity was observed in cells entering the stationary phase of growth. However, PSD activity was found to be regulated in response to soluble precursors of phospholipid biosynthesis...

  10. Metabolism of lysine-chromium complex in Saccharomyces cerevisiae

    OpenAIRE

    Karthikeyan, K. S.; Polasa, H.; Sastry, K. Sivarama; Reddy, Gopal

    2008-01-01

    Saccharomyces cerevisiae which cannot utilize lysine as a sole nitrogen source is shown to metabolize a Lysine 3 Cr3+ (1:1) complex synthesized, as a combined nitrogen and carbon source. It induces rapid uptake of lysine and prevents loss of viability, in contrast with free lysine. That complexation with trivalent chromium has the effect of profoundly influencing intracellular distribution and metabolism of the liganded amino acid is demonstrated.

  11. Genetic Basis for Saccharomyces cerevisiae Biofilm in Liquid Medium

    OpenAIRE

    Andersen, Kaj Scherz; Bojsen, Rasmus; Sørensen, Laura Gro Rejkjær; Nielsen, Martin Weiss; Lisby, Michael; Folkesson, Anders; Regenberg, Birgitte

    2014-01-01

    Biofilm-forming microorganisms switch between two forms: free-living planktonic and sessile multicellular. Sessile communities of yeast biofilms in liquid medium provide a primitive example of multicellularity and are clinically important because biofilms tend to have other growth characteristics than free-living cells. We investigated the genetic basis for yeast, Saccharomyces cerevisiae, biofilm on solid surfaces in liquid medium by screening a comprehensive deletion mutant collection in th...

  12. Quantitative analysis of the heat shock response of Saccharomyces cerevisiae.

    OpenAIRE

    Miller, M. J.; Xuong, N. H.; Geiduschek, E. P.

    1982-01-01

    Transient protein synthesis in Saccharomyces cerevisiae, after shift from 21-23 degrees C to 37 degrees C, was quantitatively analyzed. Pulse-labeled proteins were separated by two-dimensional gel electrophoresis, and autoradiograms of the gels were analyzed by a recently described method involving a computer-coupled film scanning system. In this way, the rate of incorporation of L-[35S]methionine into approximately 500 proteins was followed. The synthesis of more than 80 of these proteins wa...

  13. A mechanism of palindromic gene amplification in Saccharomyces cerevisiae

    OpenAIRE

    Rattray, Alison J.; Shafer, Brenda K.; Neelam, Beena; Strathern, Jeffrey N.

    2005-01-01

    Selective gene amplification is associated with normal development, neoplasia, and drug resistance. One class of amplification events results in large arrays of inverted repeats that are often complex in structure, thus providing little information about their genesis. We made a recombination substrate in Saccharomyces cerevisiae that frequently generates palindromic duplications to repair a site-specific double-strand break in strains deleted for the SAE2 gene. The resulting palindromes are ...

  14. Molecular Analysis of Maltotriose Transport and Utilization by Saccharomyces cerevisiae

    OpenAIRE

    Day, Rachel E.; Rogers, Peter J.; Dawes, Ian W.; Higgins, Vincent J.

    2002-01-01

    Efficient fermentation of maltotriose is a desired property of Saccharomyces cerevisiae for brewing. In a standard wort, maltotriose is the second most abundant sugar, and slower uptake leads to residual maltotriose in the finished product. The limiting factor of sugar metabolism is its transport, and there are conflicting reports on whether a specific maltotriose permease exists or whether the mechanisms responsible for maltose uptake also carry out maltotriose transport. In this study, radi...

  15. The mannoprotein of Saccharomyces cerevisiae is an effective bioemulsifier.

    OpenAIRE

    Cameron, D. R.; Cooper, D. G.; Neufeld, R. J.

    1988-01-01

    The mannoprotein which is a major component of the cell wall of Saccharomyces cerevisiae is an effective bioemulsifier. Mannoprotein emulsifier was extracted in a high yield from whole cells of fresh bakers' yeast by two methods, by autoclaving in neutral citrate buffer and by digestion with Zymolase (Miles Laboratories; Toronto, Ontario, Canada), a beta-1,3-glucanase. Heat-extracted emulsifier was purified by ultrafiltration and contained approximately 44% carbohydrate (mannose) and 17% prot...

  16. Increasing NADH oxidation reduces overflow metabolism in Saccharomyces cerevisiae

    OpenAIRE

    Vemuri, Goutham; Eiteman, M.A; McEwen, J. E.; Olsson, Lisbeth; Nielsen, Jens

    2007-01-01

    Respiratory metabolism plays an important role in energy production in the form of ATP in all aerobically growing cells. However, a limitation in respiratory capacity results in overflow metabolism, leading to the formation of byproducts, a phenomenon known as ‘‘overflow metabolism’’ or ‘‘the Crabtree effect.’’ The yeast Saccharomyces cerevisiae has served as an important model organism for studying the Crabtree effect. When subjected to increasing glycolytic fluxes under aerobic conditions, ...

  17. Engineering the Monomer Composition of Polyhydroxyalkanoates Synthesized in Saccharomyces cerevisiae

    OpenAIRE

    Zhang, Bo; Carlson, Ross; Srienc, Friedrich

    2006-01-01

    Polyhydroxyalkanoates (PHAs) have received considerable interest as renewable-resource-based, biodegradable, and biocompatible plastics with a wide range of potential applications. We have engineered the synthesis of PHA polymers composed of monomers ranging from 4 to 14 carbon atoms in either the cytosol or the peroxisome of Saccharomyces cerevisiae by harnessing intermediates of fatty acid metabolism. Cytosolic PHA production was supported by establishing in the cytosol critical ?-oxidatio...

  18. Arsenic oxide-induced thermotolerance in Saccharomyces cerevisiae.

    OpenAIRE

    Chang, E. C.; Kosman, D. J.; Willsky, G. R.

    1989-01-01

    The growth response of Saccharomyces cerevisiae to arsenite and arsenate and the relationship between the enhancement of heat shock protein (hsp) synthesis caused by these arsenic oxides and thermotolerance are reported. Arsenite and arsenate transiently inhibited cell growth and overall protein synthesis; arsenate enhanced the synthesis of the 42-, 74-, 84-, and 100-kilodalton hsps, whereas arsenite enhanced synthesis of only the 74-kilodalton hsp. The induction of these hsps reached a maxim...

  19. The enantioselective b-keto ester reductions by Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    HASSAN TAJIK

    2006-09-01

    Full Text Available The enantioselective yeast reduction of aromatic b-keto esters, by use of potassium dihydrogen phosphate, calcium phosphate (monobasic, magnesium sulfate and ammonium tartrate (diammonium salt (10:1:1:50 in water at pH 7 as a buffer for 72–120 h with 45–90 % conversion to the corresponding aromatic -hydroxy esters was achieved by means of Saccharomyces cerevisiae.

  20. An apoptotic cell cycle mutant in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Villadsen, Ingrid

    1996-01-01

    The simple eukaryote Saccharomyces cerevisiae has proved to be a useful organism for elucidating the mechanisms that govern cell cycle progression in eukaryotic cells. The excellent in vivo system permits a cell cycle study using temperature sensitive mutants. In addition, it is possible to study many genes and gene products from higher eukaryotes in Saccharomyces cerevisiae because many genes and biological processes are homologous or similar in lower and in higher eukaryotes. The highly developed methods of genetics and molecular biology greatly facilitates studies of higher eukaryotic processes.Programmmed cell death with apoptosis plays a major role in development and homeostatis in most, if not all, animal cells. Apoptosis is a morphologically distinct form of death, that requires the activation of a highly regulated suicide program. Saccharomyces cerevisiae provides a new system in which apoptosis can be studied using the novel, temperature sensitive mutant, cdc77. The cdc77 cells are defective in a G1 process, and die show the characteristc signs of apoptosis: condensation of the chromatin, degradation of the inner nuclear membrane, dilation of the space between the nuclear membranes, condensation of the cytoplasm and degradation of DNA to 50kb fragmensts. It should be noted that in yeast, in contrast to higher eukaryotes, the nuclear membrane remain intact and the chromosomes remain uncondensed and invisible during mitosis. The cdc77 mutant exhibit a defect in initiation of DNA synthesis and a much prolonged DNA synthesis under semirestrictive conditions.

  1. Genetic engineering of industrial strains of Saccharomyces cerevisiae.

    Science.gov (United States)

    Le Borgne, Sylvie

    2012-01-01

    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

  2. Saccharomyces cerevisiae en la fabricación del licor Cocuy. / Saccharomyces cerevisiae in the manufacturing of Cocuy liquor

    Scientific Electronic Library Online (English)

    F, Yegres; G, Fernández-Zeppenfeldt; CG, Padin; L, Rovero; N, Richard-Yegres.

    2003-01-01

    Full Text Available El licor "cocuy" es una bebida artesanal, producida por las comunidades rurales en el occidente de Venezuela mediante un proceso de fermentación y destilación del mosto extraído del Agave cocui. Este estudio fue enmarcado en el "Programa Agave" con el propósito de contribuir a rescatar esta activida [...] d productiva tradicional. En vista de la falta de información en relación al proceso autóctono se hicieron estudios de las levaduras fermentadoras, la optimización de la producción de etanol y la utilización del residuo de la destilación (vinaza) como medio de cultivo. Los aislados con mayor capacidad fermentativa fueron seleccionados e identificados mediante parámetros morfológicos y metabólicos. Se compararon los niveles de consumo de azúcar de las levaduras con mayor capacidad fermentativa. Se estudió el efecto de la adición del azúcar blanca comercial y/o del fosfato de amonio y en la producción de alcohol en el proceso artesanal. Las concentraciones de azúcares en el mosto se evaluaron por refractometría, y el contenido de alcohol del licor por hidrometría. La utilización de la vinaza para la producción de biomasa como un componente del medio de cultivo fue comparada con un medio sintético mediante medidas del peso seco de la biomasa. Se confirma el papel de Saccharomyces cerevisiae en el proceso fermentativo espontáneo. Los resultados in situ evidenciaron un efecto favorable de la elevación del contenido de azúcar (11 a 18°Brix) y de la adición de fosfato de amonio dibásico (0,2 g/l). En estas condiciones, el tiempo de fermentación del mosto se acortó y la producción de licor aumentó hasta un 92%. Se demostró la posibilidad de utilizar la vinaza como un componente para un medio de cultivo de esta levadura, para iniciar la fermentación y para la producción de biomasa como una fuente de nutrientes de alto valor nutritivo para aves de corral o caprinos. Se recomienda apoyar los esfuerzos para desarrollar de esta importante fuente de ingresos para los campesinos que habitan las zonas semi-áridas de los estados Falcón y Lara. Abstract in english Liquor cocuy is an alcoholic beverage produced by fermentation and a subsequent distillation process of Agave cocui juice by the communities northwest Venezuela. This study was included in the "Agave cocui Program", which purpose was to rescue this traditional productive activity. Due to the lack of [...] information about this native product fermentative yeasts, alcohol production optimization, and use of distillation residue (nasty wine) as culture medium was investigated. Isolates with the best fermentative capacity were selected and identified by morphological and metabolic studies. Sugars consumption of the yeast with highest fermentative capacity were compared. Effect of white commercial sugar and/or ammonium phosphate dibasic addition to juice was evaluated during the process. Sugars concentration was estimated with a refractometer, measurement of alcohol content of liquor with an hidrometer. The production of yeasts biomass grown in a broth with nasty wine was compared to that in a synthetic one by dry biomass weight determinations. The Saccharomyces cerevisiae role in the spontaneous fermentative process was confirmed. The results in situ proved the favorable effect of extra sugar (11 to 18°Brix) and ammonium phosphate (0,2 g/l). Fermentative period was reduced and liquor production was enhanced to 92%. Use of nasty wine was proposed to obtain yeasts biomass, as fermentation starter or as a source of high nutritive value for poultry and goat feeding. We recommend to support efforts to improve this local activity which represent an important source of income for the farmer at the semi-arid zone at the states of Falcón and Lara.

  3. Fermentation capacity of Saccharomyces cerevisiae cultures

    OpenAIRE

    Elvira Maria Bezerra de Alencar; Cristina Maria Souza-Motta; Bruno Souza Walter; Rejane Maria Pessoa Santos; Olga Martins Marques; Lusinete Aciole de Queiroz

    2009-01-01

    This work aimed at the assessment of fermentative capacity of original diploid, monocellular haploid and recuperated diploid cultures of S. cerevisiae in sterilized sugar-cane wort. Twenty eight cultures were analyzed, four being original diploids (URM-4420, Itaiquara Ferment FIT, Lallemand Ferment FLA and Wild Ferment SEL); 12 monocellular haploids from original ones and 12 recuperated diploids from the monocells. The ethanol percentage ranged from 1.7 to 6.2% and the unfermentable reducing ...

  4. Localization and cell surface anchoring of the Saccharomyces cerevisiae flocculation protein Flo1p.

    OpenAIRE

    Bony, M.; Thines-sempoux, D.; Barre, P.; Blondin, B.

    1997-01-01

    The Saccharomyces cerevisiae FLO1 gene encodes a large 1,536-amino-acid serine- and threonine-rich protein involved in flocculation. We have assessed the localization of Flo1p by immunoelectron microscopy, and in this study we show that this protein is located in the external mannoprotein layer of the cell wall, at the plasma membrane level and in the periplasm. The protein was also visualized in the endoplasmic reticulum and in the nuclear envelope, indicating that it was secreted through th...

  5. Isolation and Characterization of Mutations in the ?-Tubulin Gene of SACCHAROMYCES CEREVISIAE

    OpenAIRE

    Thomas, James H.; Neff, Norma F.; Botstein, David

    1985-01-01

    Of 173 mutants of Saccharomyces cerevisiae resistant to the antimitotic drug benomyl (BenR), six also conferred cold-sensitivity for growth and three others conferred temperature-sensitivity for growth in the absence of benomyl. All of the benR mutations tested, including the nine conditional-lethal mutations, were shown to be in the same gene. This gene, TUB2, has previously been molecularly cloned and identified as the yeast structural gene encoding ?-tubulin. Four of the conditional-letha...

  6. Daughter-specific repression of Saccharomyces cerevisiae HO: Ash1 is the commander

    OpenAIRE

    Cosma, Maria Pia

    2004-01-01

    The GATA-1-like factor Ash1 is a repressor of the HO gene, which encodes an endonuclease that is responsible for mating-type switching in the yeast Saccharomyces cerevisiae. A multi-step programme, which involves a macromolecular protein complex, the secondary structure of ASH1 mRNA and the cell cytoskeleton, enables Ash1 to asymmetrically localize to the daughter cell nucleus in late anaphase and to repress HO transcription. The resulting Ash1 activity prevents the daughter cell from switchi...

  7. AFT1: a mediator of iron regulated transcriptional control in Saccharomyces cerevisiae.

    OpenAIRE

    Yamaguchi-Iwai, Y; Dancis, A; Klausner, R D

    1995-01-01

    Using a scheme for selecting mutants of Saccharomyces cerevisiae with abnormalities of iron metabolism, we have identified a gene, AFT1, that mediates the control of iron uptake. AFT1 encodes a 78 kDa protein with a highly basic amino terminal domain and a glutamine-rich C-terminal domain, reminiscent of transcriptional activators. The protein also contains an amino terminal and a C-terminal region with 10% His residues. A dominant mutant allele of this gene, termed AFT1-1up, results in high ...

  8. Regulation of STA1 gene expression by MAT during the life cycle of Saccharomyces cerevisiae.

    OpenAIRE

    Dranginis, A. M.

    1989-01-01

    STA1 encodes a secreted glucoamylase of the yeast Saccharomyces cerevisiae var. diastaticus. Glucoamylase secretion is controlled by the mating type locus MAT; a and alpha haploid yeast cells secrete high levels of the enzyme, but a/alpha diploid cells produce undetectable amounts. It has been suggested that STA1 is regulated by MATa2 (I. Yamashita, Y. Takano, and S. Fukui, J. Bacteriol. 164:769-773, 1985), which is a MAT transcript of previously unknown function. In contrast, this work shows...

  9. Determinants for glycophospholipid anchoring of the Saccharomyces cerevisiae GAS1 protein to the plasma membrane.

    OpenAIRE

    Nuoffer, C.; Jenö, P; Conzelmann, A; Riezman, H

    1991-01-01

    A 125-kDa glycoprotein exposed on the surface of Saccharomyces cerevisiae cells belongs to a class of eucaryotic membrane proteins anchored to the lipid bilayer by covalent linkage to an inositol-containing glycophospholipid. We have cloned the gene (GAS1) encoding the 125-kDa protein (Gas1p) and found that the function of Gas1p is not essential for cell viability. The nucleotide sequence of GAS1 predicts a 60-kDa polypeptide with a cleavable N-terminal signal sequence, potential sites for N-...

  10. N myristylation of the human immunodeficiency virus type 1 gag polyprotein precursor in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    A semisynthetic gene precisely encoding the 502 amino acids of the human immunodeficiency virus type 1 gag precursor (Pr53gag) was expressed in the yeast Saccharomyces cerevisiae. Amino acid sequence analysis of the recombinant Pr53gag showed that the amino terminus was fully blocked. Labeling of Pr53gag with [3H]myristic acid demonstrated that, as with Pr53gag isolated from virus-infected cells, the yeast-derived protein was demethionylated and N myristylated on glycine, the second amino acid residue

  11. Saccharomyces cerevisiae: a nomadic yeast with no niche?

    Science.gov (United States)

    Goddard, Matthew R; Greig, Duncan

    2015-05-01

    Different species are usually thought to have specific adaptations, which allow them to occupy different ecological niches. But recent neutral ecology theory suggests that species diversity can simply be the result of random sampling, due to finite population sizes and limited dispersal. Neutral models predict that species are not necessarily adapted to specific niches, but are functionally equivalent across a range of habitats. Here, we evaluate the ecology of Saccharomyces cerevisiae, one of the most important microbial species in human history. The artificial collection, concentration and fermentation of large volumes of fruit for alcohol production produce an environment in which S. cerevisiae thrives, and therefore it is assumed that fruit is the ecological niche that S. cerevisiae inhabits and has adapted to. We find very little direct evidence that S. cerevisiae is adapted to fruit, or indeed to any other specific niche. We propose instead a neutral nomad model for S. cerevisiae, which we believe should be used as the starting hypothesis in attempting to unravel the ecology of this important microbe. PMID:25725024

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1982-05-01

    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).

  13. Cloning and expression of a Saccharomyces diastaticus glucoamylase gene in Saccharomyces cerevisiae and Schizosaccharomyces pombe.

    OpenAIRE

    Erratt, J A; Nasim, A

    1986-01-01

    A recombinant plasmid pool of the Saccharomyces diastaticus genome was constructed in plasmid YEp13 and used to transform a strain of Saccharomyces cerevisiae. Six transformants were obtained which expressed amylolytic activity. The plasmids each contained a 3.9-kilobase (kb) BamHI fragment, and all of these fragments were cloned in the same orientations and had identical restriction maps, which differed from the map of the STA1 gene (I. Yamashita and S. Fukui, Agric. Biol. Chem. 47:2689-2692...

  14. Ultradian oscillators of the circadian clock in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Sara S. Dick

    2013-02-01

    Full Text Available The yeast, Saccharomyces cerevisiae, has an ENOX1 activity with a period length of 24 min similar to that of other eukaryotes. In contrast to other eukaryotes, however, Saccharomyces cerevisiae has a second ENOX1-like activity with a period length of 25 min. The latter is distinguishable from the traditional ENOX1 on the basis of the longer period length along with resistance to an ENOX1 inhibitor, simalikalactone D, and failure to be phased by melatonin. In addition, two periods are apparent in measurements of oxygen consumption indicating that the consumption of oxygen to water occurs independently by homodimers of both of the two forms of ENOX. Based on the measurements of glyceraldehyde-3- phosphate dehydrogenase, S. cerevisiae exhibits circadian activity maxima at 24 and 25 h together with a 40 h period possibly representing the 40 min metabolic rhythm of yeast not observed in our measurement of oxygen consumption and normally observed only with continuous cultures. The findings are indicative of at least three independent time-keeping systems being operative in a single cell. 

  15. A polyploid population of Saccharomyces cerevisiae with separate sexes (dioecy).

    Science.gov (United States)

    Al Safadi, Rim; Weiss-Gayet, Michèle; Briolay, Jérôme; Aigle, Michel

    2010-09-01

    Saccharomyces cerevisiae has proved to be an interesting model for studies of evolution, with whole-genome duplication shown to have played an important role in the evolution of this species. This phenomenon depends on the formation of a transient stable polyploid state. Previous studies have reported polyploidy to be an unstable state in yeast, but here, we describe a polyploid population of S. cerevisiae. The evolution of higher eukaryotes has also involved the development of different systems of sexual reproduction, the choice between self-fertilization and out-crossing becoming a key issue. Saccharomyces cerevisiae is a hermaphrodite eukaryote, despite the theoretical genetic disadvantages of this strategy, in which self-fertilization occurs. We describe, for the first time, a near-dioecious (with separate sexes) population in this species. Mating type and the MAT locus display complex segregations. Essentially, each strain produces, by meiosis, spores of only one mating type: mata or matalpha. Moreover, strains are heterothallic, and diploid nonmating clones generated from a single spore do not sporulate. These three properties limit self-fertilization and strongly favour out-crossing. We suggest that the shift in sexual strategy, from hermaphroditism to dioecy, is specific to the brewing process, which overcomes the sexual isolation probably found in natural biotopes. PMID:20662936

  16. Construction of killer industrial yeast Saccharomyces cerevisiae HAU-1 and its fermentation performance

    Scientific Electronic Library Online (English)

    Bijender K., Bajaj; S., Sharma.

    2010-06-01

    Full Text Available Saccharomyces cerevisiae HAU-1, a time tested industrial yeast possesses most of the desirable fermentation characteristics like fast growth and fermentation rate, osmotolerance, high ethanol tolerance, ability to ferment molasses, and to ferment at elevated temperatures etc. However, this yeast was [...] found to be sensitive against the killer strains of Saccharomyces cerevisiae. In the present study, killer trait was introduced into Saccharomyces cerevisiae HAU-1 by protoplast fusion with Saccharomyces cerevisiae MTCC 475, a killer strain. The resultant fusants were characterized for desirable fermentation characteristics. All the technologically important characteristics of distillery yeast Saccharomyces cerevisiae HAU-1 were retained in the fusants, and in addition the killer trait was also introduced into them. Further, the killer activity was found to be stably maintained during hostile conditions of ethanol fermentations in dextrose or molasses, and even during biomass recycling.

  17. Construction of killer industrial yeast Saccharomyces cerevisiae HAU-1 and its fermentation performance

    Directory of Open Access Journals (Sweden)

    Bijender K. Bajaj

    2010-06-01

    Full Text Available Saccharomyces cerevisiae HAU-1, a time tested industrial yeast possesses most of the desirable fermentation characteristics like fast growth and fermentation rate, osmotolerance, high ethanol tolerance, ability to ferment molasses, and to ferment at elevated temperatures etc. However, this yeast was found to be sensitive against the killer strains of Saccharomyces cerevisiae. In the present study, killer trait was introduced into Saccharomyces cerevisiae HAU-1 by protoplast fusion with Saccharomyces cerevisiae MTCC 475, a killer strain. The resultant fusants were characterized for desirable fermentation characteristics. All the technologically important characteristics of distillery yeast Saccharomyces cerevisiae HAU-1 were retained in the fusants, and in addition the killer trait was also introduced into them. Further, the killer activity was found to be stably maintained during hostile conditions of ethanol fermentations in dextrose or molasses, and even during biomass recycling.

  18. Expression of the E.coli pntA and pntB genes encoding nicotinamide nucleotide transhydrogenase in Saccharomyces cerevisiae and its effect on product formation during anaerobic glucose fermentation

    DEFF Research Database (Denmark)

    Anderlund, M.; Nissen, Torben Lauesgaard

    1999-01-01

    We studied the physiological effect of the interconversion between the NAD(H) and NADP(H) coenzyme systems in recombinant Saccharomyces cerevisiae expressing the membrane-bound transhydrogenase from Escherichia coli. Our objective was to determine if the membrane-bound transhydrogenase could work in reoxidation of NADH to NAD(+) in S. cerevisiae and thereby reduce glycerol formation during anaerobic fermentation. Membranes isolated from the recombinant strains exhibited reduction of 3-acetylpyridine-NAD(+) by NADPH and by NADH in the presence of NADP(+), which demonstrated that an active enzyme was present. Unlike the situation in E. coli, however, most of the transhydrogenase activity was not present in the yeast plasma membrane; rather, the enzyme appeared to remain localized in the membrane of the endoplasmic reticulum. During anaerobic glucose fermentation we observed an increase in the formation of 2-oxoglutarate, glycerol, and acetic acid in a strain expressing a high level of transhydrogenase, which indicated that increased NADPH consumption and NADH production occurred. The intracellular concentrations of NADH, NAD(+) NADPH, and NADP(+) were measured in cells expressing transhydrogenase. The reduction of the NADPH pool indicated that the transhydrogenase transferred reducing equivalents from NADPH to NAD(+).

  19. Cloning and DNA sequence analysis of the glucose transporter gene2 from Iranian Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Saleh Amiri

    2012-12-01

    Full Text Available Introduction: Saccharomyces cerevisiae has 20 genes that encode hexose transporter proteins including HXT1 to HXT17, GAL2, SNF3 and RGT2. Among these gene families, seven genes (HXT1-HXT7 have important role in alcohol production. The aim of this study was the identification and isolation of HXT2 gene from Saccharomyces cerevisiae genome by PCR technique and cloning into vector containing suitable expression promoter in order to design expression vector as a basis to produce recombinant yeast by transformation.Materials and Methods: After designing specific oligonucleotides primers, fragment gene amplified by PCR. Gene HXT2 inserted into pTZ57R vector by restriction enzymes EcoRI and HindIII and T4 ligase. After transformation of pTZ57R/THXT2 into E.coli, plasmid recombinant analysis considered. The final further analysis by restriction enzymes digestion and software were evaluated.Results: HXT2 gene isolated from pTZ57R/THXT2 has correct size in agarose gel electrophoresis. Electrophoresis analysis showed that this gene has correct size on agarose gel. Software study showed that this gene encode proteins with 59.84 KDa molecular weight having 541 amino acids with isoelectric point 8.3.Conclusion: HXT2 gene by PCR optimization from saccharomyces cerevisiae was isolated and cloned into prokaryotic host. This is the first report of isolation and cloning of this gene by using genetic engineering technique in IRAN that can be used for cloning into suitable expression vector to improve alcohol fermentation yield.

  20. ISOTERMAS DE ADSORÇÃO DE CÁDMIO POR Saccharomyces cerevisiae ISOTHERMS OF CADMIUM ADSORPTION BY Saccharomyces cerevisae

    Directory of Open Access Journals (Sweden)

    Silvana ALBERTINI

    2001-08-01

    Full Text Available Com o objetivo de determinar as isotermas de adsorção de cádmio por Saccharomyces cerevisiae, foram utilizados os sais cloreto e nitrato de cádmio nas concentrações de 5, 10, 20, 40, 60, 80 e 100mg L-1. A biomassa foi produzida a partir de uma cultura "starter"de Saccharomyces cerevisiae IZ 1904. Após o contato de 16h entre o microrganismo e as soluções em estudo, a biomassa foi separada por centrifugação e o teor de cádmio residual foi determinado no sobrenadante por espectrofotometria de absorção atômica. Para os dois sais empregados foi observado um acúmulo crescente de cádmio nas concentrações de 5, 10, 20 e 40mg L-1. Nas concentrações de 60, 80 e 100mg L-1 foi observado que a levedura acumulou teores menores do metal, evidenciando danos na parede celular, nem sempre acompanhados de iguais danos da membrana citoplasmática, tais alterações da parede visualizadas por microscopia eletrônica de varredura.With the objective of determining the isotherms of cadmium the adsorption by Saccharomyces cerevisiae, the chloride and nitrate salts were used in the concentrations of 5, 10, 20, 40, 60, 80, and 100mg L-1. The biomass was produced from a starter culture of Saccharomyces cerevisiae IZ 1904. After a 16h contact between the microrganism and solutions of study the biomass was separated by a centrifuge and the cadmium residue content was determined at the supernatant by atomic adsorption spectrophotometry. For the two salts used a growing accumulation of cadmium was observed at concentrations of 5, 10, 20, and 40mg L-1. In the concentrations of 60, 80 and 100mg L-1 a decreasing of the accumulation of the metal was observed, evidencing damages of the cellular wall, which they're not accompanied always by damages of the citoplasmatic membrane, visualized by scanning electron microscopy.

  1. Saccharomyces cerevisiae and non-Saccharomyces yeasts in grape varieties of the São Francisco Valley

    Scientific Electronic Library Online (English)

    Camila M.P.B.S. de, Ponzzes-Gomes; Dângelly L.F.M. de, Mélo; Caroline A., Santana; Giuliano E., Pereira; Michelle O.C., Mendonça; Fátima C.O., Gomes; Evelyn S., Oliveira; Antonio M., Barbosa Jr; Rita C., Trindade; Carlos A., Rosa.

    2014-06-01

    Full Text Available The aims of this work was to characterise indigenous Saccharomyces cerevisiae strains in the naturally fermented juice of grape varieties Cabernet Sauvignon, Grenache, Tempranillo, Sauvignon Blanc and Verdejo used in the São Francisco River Valley, northeastern Brazil. In this study, 155 S. cerevisi [...] ae and 60 non-Saccharomyces yeasts were isolated and identified using physiological tests and sequencing of the D1/D2 domains of the large subunit of the rRNA gene. Among the non-Saccharomyces species, Rhodotorula mucilaginosa was the most common species, followed by Pichia kudriavzevii, Candida parapsilosis, Meyerozyma guilliermondii, Wickerhamomyces anomalus, Kloeckera apis, P. manshurica, C. orthopsilosis and C. zemplinina. The population counts of these yeasts ranged among 1.0 to 19 x 10(5) cfu/mL. A total of 155 isolates of S. cerevisiae were compared by mitochondrial DNA restriction analysis, and five molecular mitochondrial DNA restriction profiles were detected. Indigenous strains of S. cerevisiae isolated from grapes of the São Francisco Valley can be further tested as potential starters for wine production.

  2. Protein expression of saccharomyces cerevisiae in response to uranium exposure

    International Nuclear Information System (INIS)

    Protein expression of Saccharomyces cerevisiae grown in the medium containing 238U (VI) and 233U (VI) was examined by two-dimensional gel electrophoresis. Saccharomyces cerevisiae of BY4743 was grown in yeast nitrogen base medium containing glucose and glycerol 2-phosphate and 238U of 0, 2.0, and 5.0 x 10-4 M or 233U of 2.5 x 10-6 M (radioactivity was higher by 350 times than 2.0 x 10-4 M 238U) and 5.0 x 10-6 M for 112 h at 30 degC. The growth of Saccharomyces cerevisiae was monitored by measuring OD600 at 112 h after the inoculation. Uranium concentrations in the media also were measured by radiometry using a liquid scintillation counter. The growths of the yeast grown in the above media were in the following order: control>2.5 x 10-6 M 233U>2.0 x 10-4 M 238U>5.0 x 10-6 M 233U>5.0 x 10-4 M 238U. This result indicated that not only radiological but also chemical effect of U reduced the growth of the yeast. The concentrations of U in the medium containing 238U or 233U decreased, suggesting U accumulation by the yeast cells. The 2-D gel electrophoresis analysis showed the appearance of several spots after exposure to 238U or to 233U but not in the control containing no uranium. These results show that the yeast cells exposed to U ex show that the yeast cells exposed to U express several specific proteins. (author)

  3. Applied systems biology - vanillin production in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Strucko, Tomas; Eriksen, Carsten

    2012-01-01

    Vanillin is the most important aroma compound based on market value, and natural vanillin is extracted from the cured seed pods of the Vanilla orchid. Most of the world’s vanillin, however, is obtained by chemical synthesis from petrochemicals or wood pulp lignins. As an alternative, de novo biosynthesis of vanillin in baker’s yeast Saccharomyces cerevisiae was recently demonstrated by successfully introducing the metabolic pathway for vanillin production in yeast. Nevertheless, the amount of vanillin produced in this S. cerevisiae strain is insufficient for commercial production and improvements need to be done. We have introduced the genes necessary for vanillin production in an identical manner in two different yeast strains S288c and CEN.PK,where comprehensive – omics datasets are available, hence, allowing vanillin production in the two strain backgrounds to be evaluated and compared in a systems biology setting.

  4. Understanding the 3-hydroxypropionic acid tolerance mechanism in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Kildegaard, Kanchana Rueksomtawin; Juncker, Agnieszka

    2013-01-01

    3-Hydroxypropionic acid (3HP) is an important platform chemical that can be converted into other valuable chemicals such as acrylic acid and its derivatives that are used in baby diap ers, various plastics, and paints. With the oil and gas resources becoming limiting, biotechnolo gy offers a sustainable alternative for production of acrylic acid from renewable feedstocks. We are establishing Saccharomyces cerevisiae as an alternative host for 3HP production. However, 3HP also inhibits yeast grow th at level well below what is desired for commercial applications. Therefore, we are aiming to improve 3HP tolerance in S. cerevisiae by applying adaptive evolution approach. We have generated yeast strains with sign ificantly improved capacity for tolerating 3HP when compared to the wild-type. We will present physiolo gical characterization, genome re-sequencing, and transcriptome analysis of the evolved strains. Conseq uently, mechanism underlying 3HP tolerance will be investigated.

  5. Hierarchical Bayesian Modelling for Saccharomyces cerevisiae population dynamics.

    Science.gov (United States)

    Spor, Aymé; Dillmann, Christine; Wang, Shaoxiao; de Vienne, Dominique; Sicard, Delphine; Parent, Eric

    2010-08-15

    Hierarchical Bayesian Modelling is powerful however under-used to model and evaluate the risks associated with the development of pathogens in food industry, to predict exotic invasions, species extinctions and development of emerging diseases, or to assess chemical risks. Modelling population dynamics of Saccharomyces cerevisiae considering its biodiversity and other sources of variability is crucial for selecting strains meeting industrial needs. Using this approach, we studied the population dynamics of S. cerevisiae, the domesticated yeast, widely encountered in food industry, notably in brewery, vinery, bakery and distillery. We relied on a logistic equation to estimate the key variables of population growth, but we took also into account factors able to affect them, namely environmental effects, genetic diversity and measurement errors. Our probabilistic approach allowed us: (i) to model the dynamical behaviour of strains in a given condition under some uncertainty, (ii) to measure environmental effects and (iii) to evaluate genetic variability of the growth key variables. PMID:20576304

  6. Effect of environmental stress on radiation response of Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    We have investigated the effect of pH shock and oxidative stress (H2O2 effect) both separately and together on the response of Saccharomyces cerevisiae exposed to UV and gamma radiations for one hour. Exposure to these environmental stresses resulted in S. cerevisiae cells acquiring resistance to UV radiation. Presence of cycloheximide (a know protein synthesis inhibitor) during stress inhibited the acquired UV resistance. The increased UV resistance is apparently mediated through nucleotide excision repair as the stress exposure to rad3 mutants (defective in nucleotide excision repair) do not have any effect on UV response. Both types of stresses used probably follow the same path of induction of radioresistance as the effect of both of them is nonadditive. In the strains used in our study stress exposure dose not have any significant effect on gamma radiation response. (author)

  7. Expression of Saccharomyces cerevisiae ?-glucoside transporters under different growth conditions

    Scientific Electronic Library Online (English)

    S. L., Alves Jr.; J. M., Thevelein; B. U., Stambuk.

    2014-03-01

    Full Text Available Important biotechnological processes depend on the efficient fermentation by Saccharomyces cerevisiae yeasts of starch hydrolysates rich in maltose and maltotriose. The rate-limiting step for fermentation of these ?-glucosides is the transport across the plasma membrane of the cells. In order to con [...] tribute to a better understanding of maltose and maltotriose metabolism by S. cerevisiae, we analyzed the expression of the main ? glucoside transporter genes in two different yeast strains grown on media with glucose, maltose or maltotriose as carbon source. Although both yeast strains have higher ?glucoside transport activity during growth on maltotriose, our results show similar expression levels of the analyzed genes on either maltose or maltrotriose media. Thus, our results indicate that, although the transport capacity of maltotriose grown cells is higher than that of maltose grown cells, maltotriose cannot be considered a better inducer of ?glucoside transporter genes.

  8. Glucose- and nitrogen sensing and regulatory mechanisms in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    RØdkaer, Steven V; Færgeman, Nils J.

    2014-01-01

    Pro- and eukaryotic cells are constantly challenged by varying concentrations of nutrients in their environment. Perceiving and adapting to such changes are therefore crucial for cellular viability. Thus, numerous specialized cellular receptors continuously sense and react to the availability of nutrients such as glucose and nitrogen. When stimulated, these receptors initiate various cellular signaling pathways, which in concert constitute a complex regulatory network. To ensure a highly specific response, these pathways and networks cross-communicate with each other and are regulated at several steps and by numerous different regulators. As numerous of these regulating proteins, biochemical mechanisms, and cellular pathways are evolutionary conserved, complex biochemical information relevant to humans can be obtained by studying simple organisms. Thus, the yeast Saccharomyces cerevisiae has been recognized as a powerful model system to study fundamental biochemical processes. In the present review, we highlight central signaling pathways and molecular circuits conferring nitrogen- and glucose sensing in S. cerevisiae.

  9. Use of Saccharomyces cerevisiae in radioactive waste treatment

    International Nuclear Information System (INIS)

    Waste management plays an important role in reducing the volume of radioactive waste streams, minimizing the cost of the final disposal and the impact on the environment. In this context, new research should focus on the development of simpler and cheaper techniques which may improve the waste processing. The use of biomass in processes concerned with the removal of heavy metals and radionuclides offers significant potential in the treatment of waste-liquid streams. Saccharomyces cerevisiae is well known for its capacity of heavy metals biosorption and it also has the additional advantages such as easy availability and the possibility of genetic manipulation. The aim of this work is to study the potential of the free cell and immobilized S. cerevisiae in bentonite in the removal Americium-241 from radioactive liquid streams produced by Radioactive Waste Laboratory of Nuclear and Energy Research Institute (IPEN-CNEN/SP). (author)

  10. Glucose sensing and signaling in Saccharomyces cerevisiae through the Rgt2 glucose sensor and casein kinase I

    OpenAIRE

    Moriya, Hisao; Johnston, Mark

    2004-01-01

    The yeast Saccharomyces cerevisiae senses glucose through two transmembrane glucose sensors, Snf3 and Rgt2. Extracellular glucose causes these sensors to generate an intracellular signal that induces expression of HXT genes encoding glucose transporters by inhibiting the function of Rgt1, a transcriptional repressor of HXT genes. We present the following evidence that suggests that the glucose sensors are coupled to the membrane-associated protein kinase casein kinase I (Yck1). (i) Overexpres...

  11. Induction of homologous recombination in mammalian chromosomes by using the I-SceI system of Saccharomyces cerevisiae.

    OpenAIRE

    Choulika, A; A. Perrin; Dujon, B; Nicolas, J F

    1995-01-01

    The mitochondrial intron-encoded endonuclease I-SceI of Saccharomyces cerevisiae has an 18-bp recognition sequence and, therefore, has a very low probability of cutting DNA, even within large genomes. We demonstrate that double-strand breaks can be initiated by the I-SceI endonuclease at a predetermined location in the mouse genome and that the breaks can be repaired with a donor molecule homologous regions flanking the breaks. This induced homologous recombination is approximately 2 orders o...

  12. Homofermentative Lactate Production Cannot Sustain Anaerobic Growth of Engineered Saccharomyces cerevisiae: Possible Consequence of Energy-Dependent Lactate Export:

    OpenAIRE

    Van Maris, A.J.; Winkler, A.A.; Porro, D.; Dijken, J.P. van; Pronk, J T

    2004-01-01

    Due to a growing market for the biodegradable and renewable polymer polylactic acid, the world demand for lactic acid is rapidly increasing. The tolerance of yeasts to low pH can benefit the process economy of lactic acid production by minimizing the need for neutralizing agents. Saccharomyces cerevisiae (CEN.PK background) was engineered to a homofermentative lactate-producing yeast via deletion of the three genes encoding pyruvate decarboxylase and the introduction of a heterologous lactate...

  13. NDT80 and the Meiotic Recombination Checkpoint Regulate Expression of Middle Sporulation-Specific Genes in Saccharomyces cerevisiae

    OpenAIRE

    Hepworth, Shelley R.; Friesen, Helena; Segall, Jacqueline

    1998-01-01

    Distinct classes of sporulation-specific genes are sequentially expressed during the process of spore formation in Saccharomyces cerevisiae. The transition from expression of early meiotic genes to expression of middle sporulation-specific genes occurs at about the time that cells exit from pachytene and form the meiosis I spindle. To identify genes encoding potential regulators of middle sporulation-specific gene expression, we screened for mutants that expressed early meiotic genes but fail...

  14. Regulation of the Premiddle and Middle Phases of Expression of the NDT80 Gene during Sporulation of Saccharomyces cerevisiae

    OpenAIRE

    Pak, Julia; Segall, Jacqueline

    2002-01-01

    The NDT80 gene of Saccharomyces cerevisiae, which encodes a global activator of transcription of middle sporulation-specific genes, is first expressed after the activation of early meiotic genes but prior to activation of middle sporulation-specific genes. Both upstream repression sequence 1 (URS1) and mid-sporulation element (MSE) sites are present in the promoter region of the NDT80 gene; these elements have been shown previously to contribute to the regulation of expression of early and mi...

  15. Production of a thermostable fumarase in Saccharomyces cerevisiae for the bioconversion of fumarate to L-malate

    OpenAIRE

    Signorell, Coralie

    2011-01-01

    The gene encoding fumarase (fum) from Thermus thermophilus was expressed in yeast Saccharomyces cerevisiae. The recombinant cells were heated at 70°C to inactivate indigenous enzymes and used for the bioconversion of fumaric acid to L-malic acid. By heating the host cells at 70°C, substrate is able to go across the heat-damaged membrane of the microorganism and a desired product can be formed. This new concept, called Synthetic Metabolic Engineering (SME), has already been applied successfu...

  16. Key features of the two-intron Saccharomyces cerevisiae gene SUS1 contribute to its alternative splicing

    OpenAIRE

    Hossain, Munshi Azad; Rodriguez, Caitlin M.; Johnson, Tracy L.

    2011-01-01

    Alternative pre-mRNA splicing allows dramatic expansion of the eukaryotic proteome and facilitates cellular response to changes in environmental conditions. The Saccharomyces cerevisiae gene SUS1, which encodes a protein involved in mRNA export and histone H2B deubiquitination, contains two introns; non-canonical sequences in the first intron contribute to its retention, a common form of alternative splicing in plants and fungi. Here we show that the pattern of SUS1 splicing changes in respon...

  17. A novel member of the split ??? fold: Solution structure of the hypothetical protein YML108W from Saccharomyces cerevisiae

    OpenAIRE

    Pineda-lucena, Antonio; Jack C. C. Liao; Cort, John R.; Yee, Adelinda; Kennedy, Michael A; Edwards, Aled M.; Cheryl H. Arrowsmith

    2003-01-01

    As part of the Northeast Structural Genomics Consortium pilot project focused on small eukaryotic proteins and protein domains, we have determined the NMR structure of the protein encoded by ORF YML108W from Saccharomyces cerevisiae. YML108W belongs to one of the numerous structural proteomics targets whose biological function is unknown. Moreover, this protein does not have sequence similarity to any other protein. The NMR structure of YML108W consists of a four-stranded ?-sheet with strand...

  18. Magnetically altered ethanol fermentation capacity of Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Galonja-Corghill Tamara

    2009-01-01

    Full Text Available We studied the effect of static magnetic fields on ethanol production by yeast Saccharomyces cerevisiae 424A (LNH-ST using sugar cane molasses during the fermentation in an enclosed bioreactor. Two static NdFeB magnets were attached to a cylindrical tube reactor with their opposite poles (north to south, creating 150 mT magnetic field inside the reactor. Comparable differences emerged between the results of these two experimental conditions. We found ethanol productivity to be 15% higher in the samples exposed to 150 mT magnetic field.

  19. Properties of the transcriptional enhancer in Saccharomyces cerevisiae telomeres.

    OpenAIRE

    Runge, K W; Zakian, V A

    1990-01-01

    Saccharomyces cerevisiae chromosomes end with the sequence C2-3A(CA)1-4, commonly abbreviated as C1-3A. These sequences can function as upstream activators of transcription (UAS's) when placed in front of a CYC1-lacZ fusion gene. When C1-3A sequences are placed between the GAL1,10 UAS and the CYC1-lacZ fusion, the C1-3A UAS still functions and the amount of beta-galactosidase produced in cells grown on glucose is as much or more than that for cells grown on either glycerol medium, or cells gr...

  20. Saccharomyces cerevisiae HSP70 heat shock elements are functionally distinct.

    OpenAIRE

    Young, M. R.; Craig, E. A.

    1993-01-01

    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 t...

  1. Functional evaluation of serine 252 of Saccharomyces cerevisiae phosphoenolpyruvate carboxykinase.

    Science.gov (United States)

    Castillo, Daniel; Sepúlveda, Carolina; Cardemil, Emilio; Jabalquinto, Ana M

    2009-02-01

    Saccharomyces cerevisiae phosphoenolpyruvate (PEP) carboxykinase mutant Ser252Ala, affecting the conserved Walker A serine residue, was characterized to elucidate the role of this serine residue. The substitution did not result in changes in the protein structure, as indicated by circular dichroism, intrinsic fluorescence spectroscopy, and gel-exclusion chromatography. Kinetic analysis of the mutated enzyme in both directions of the main reaction and in the two secondary reactions showed an approximately 50-fold increase in apparent K(m) for oxaloacetate with minor alterations in the other kinetic parameters. These results show that the hydroxyl group of serine 252 is required for proper oxaloacetate interaction. PMID:18996167

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

    OpenAIRE

    Naumovski, L; Friedberg, E.C.

    1984-01-01

    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...

  3. Postreplication Repair Inhibits CAG · CTG Repeat Expansions in Saccharomyces cerevisiae?

    OpenAIRE

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

    2006-01-01

    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 postreplication repair (PRR). Like srs2 mutants, expansions are elevated in rad18 and rad5 mutants, as well as the PRR-specific PCNA alleles pol30-K164R and pol30-K127/164R. Epistasis analy...

  4. Radioimmunoassay for yeast killer toxin from Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    A radioimmunoassay was developed for the K1 killer toxin from strain T158C/S14a of Saccharomyces cerevisiae. Iodine 125-labelled toxin was made to a specific activity of 100 ?Ci/mg of protein. Antibody to purified toxin was prepared in rabbits using toxin cross-linked to itself. These antibodies, partially purified by 50 percent ammonium sulfate precipitation and Sepharose CL-6B column chromatography, produced one precipitation band with killer toxin and bound 125I-labelled toxin in a radioimmunoassay. The antibody preparation also bound with the toxins from another K1 killer, A364A, and three chromosomal superkiller mutants derived from it. (auth)

  5. Identification and Characterization of Mutations Affecting Sporulation in Saccharomyces Cerevisiae

    OpenAIRE

    Smith, L. M.; Robbins, L. G.; Kennedy, A.; Magee, P. T.

    1988-01-01

    Mutations affecting the synthesis of the sporulation amyloglucosidase were isolated in a homothallic strain of Saccharomyces cerevisiae, SCMS7-1. Two were found, both of which were deficient in sporulation at 34°. One, SL484, sporulated to 50% normal levels at 30° but less than 5% at 34° or 22°. The other, SL641, failed to sporulate at any temperature. Both mutants were blocked before premeiotic DNA synthesis, and both complemented spo1, spo3, and spo7. Genetic analysis of the mutation in...

  6. Nitrogen catabolite repression of asparaginase II in Saccharomyces cerevisiae.

    OpenAIRE

    Dunlop, P. C.; Meyer, G. M.; Roon, R. J.

    1980-01-01

    The biosynthesis of asparaginase II in Saccharomyces cerevisiae is subject to strong catabolite repression by a variety of nitrogen compounds. In the present study, asparaginase II synthesis was examined in a wild-type yeast strain and in strains carrying gdhA, gdhCR, or gdhCS mutations. The following effects were observed: (i) In the wild-type strain, the biosynthesis of asparaginase II was strongly repressed when either 10 mM ammonium sulfate or various amino acids (10 mM) served as the sou...

  7. Saccharomyces cerevisiae MutL? IS A MISMATCH REPAIR ENDONUCLEASE*

    OpenAIRE

    Kadyrov, Farid A.; Holmes, Shannon F.; Arana, Mercedes E.; Lukianova, Olga A.; O’DONNELL, MIKE; Kunkel, Thomas A.; Modrich, Paul

    2007-01-01

    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) meth...

  8. Induction of polypeptides in Saccharomyces cerevisiae after ultraviolet irradiation

    International Nuclear Information System (INIS)

    Alterations in the synthesis of proteins following exposure of Saccharomyces cerevisiae to UV light were investigated using radioactive labelling and two dimensional electrophoresis. UV-irradiation induced the synthesis of various proteins. Among them the analogue of the RecA protein of Escherichia coli (Angulo et al. 1985) and two other polypeptides (34 Kd and 35 Kd, pI 5.8) were observed in all four strains analyzed namely two DNA-repair deficient (rad?) strains : (rad6-1 and pso2-1) and their isogenic wild type RAD+ strains

  9. Fermentation capacity of Saccharomyces cerevisiae cultures

    Directory of Open Access Journals (Sweden)

    Elvira Maria Bezerra de Alencar

    2009-08-01

    Full Text Available This work aimed at the assessment of fermentative capacity of original diploid, monocellular haploid and recuperated diploid cultures of S. cerevisiae in sterilized sugar-cane wort. Twenty eight cultures were analyzed, four being original diploids (URM-4420, Itaiquara Ferment FIT, Lallemand Ferment FLA and Wild Ferment SEL; 12 monocellular haploids from original ones and 12 recuperated diploids from the monocells. The ethanol percentage ranged from 1.7 to 6.2% and the unfermentable reducing sugars from 0.45 to 0.50g/100mL. The highest ethanol percentages were produced by the monocellular cultures URM-MH3 (4.8% in 12 h and SEL-MH1 (6.2% in 24 h, corresponding to the productivity values of 3.15 and 2.03 g.L-1.h-1, respectively. The recuperated diploids did not present detectable ethanol content by the method used. The results showed that depending on the physiological state of the yeasts, being in the diploid or monocellular haploid form, different percentages of ethanol could be produced.Este trabalho teve como objetivo avaliar culturas de S. cerevisiae diplóides originais, haplóides monocelulares e diplóides recuperados, quanto à capacidade de fermentar em mosto de cana-deaçúcar esterilizado. Vinte e oito culturas foram analisadas, sendo 4 diplóides originais (URM-4420, Fermento Itaiquara FIT, Fermento Lallemand FLA e Fermento Selvagem SEL; 12 haplóides monocelulares provenientes das originais e 12 diplóides recuperados a partir das monocelulares. O percentual de etanol produzido variou de 1,70% a 6,20% e os açúcares redutores infermentescíveis de 0,45 g/100mL a 0,50 g/100mL. Os maiores percentuais de etanol foram produzidos pelas culturas haplóides monocelulares URM-MH3 (4,80%, em 12 horas e SEL-MH1 (6,20%, em 24 horas, correspondendo a valores de produtividade 3,15g.L-1.h-1 e 2,03 g.L-1.h-1 , respectivamente. Os diplóides recuperados não apresentaram teores de etanol detectáveis pelo método utilizado. Os resultados mostram que, dependendo do estado fisiológico das leveduras, seja na forma diplóide ou haplóide monocelular, podem ser produzidos diferentes percentuais de etanol.

  10. Fermentation capacity of Saccharomyces cerevisiae cultures

    Scientific Electronic Library Online (English)

    Elvira Maria Bezerra de, Alencar; Cristina Maria de, Souza-Motta; Bruno Souza, Walter; Rejane Maria Pessoa, Santos; Olga Martins, Marques; Lusinete Aciole de, Queiroz.

    2009-08-01

    Full Text Available Este trabalho teve como objetivo avaliar culturas de S. cerevisiae diplóides originais, haplóides monocelulares e diplóides recuperados, quanto à capacidade de fermentar em mosto de cana-deaçúcar esterilizado. Vinte e oito culturas foram analisadas, sendo 4 diplóides originais (URM-4420, Fermento It [...] aiquara FIT, Fermento Lallemand FLA e Fermento Selvagem SEL); 12 haplóides monocelulares provenientes das originais e 12 diplóides recuperados a partir das monocelulares. O percentual de etanol produzido variou de 1,70% a 6,20% e os açúcares redutores infermentescíveis de 0,45 g/100mL a 0,50 g/100mL. Os maiores percentuais de etanol foram produzidos pelas culturas haplóides monocelulares URM-MH3 (4,80%), em 12 horas e SEL-MH1 (6,20%), em 24 horas, correspondendo a valores de produtividade 3,15g.L-1.h-1 e 2,03 g.L-1.h-1 , respectivamente. Os diplóides recuperados não apresentaram teores de etanol detectáveis pelo método utilizado. Os resultados mostram que, dependendo do estado fisiológico das leveduras, seja na forma diplóide ou haplóide monocelular, podem ser produzidos diferentes percentuais de etanol. Abstract in english This work aimed at the assessment of fermentative capacity of original diploid, monocellular haploid and recuperated diploid cultures of S. cerevisiae in sterilized sugar-cane wort. Twenty eight cultures were analyzed, four being original diploids (URM-4420, Itaiquara Ferment FIT, Lallemand Ferment [...] FLA and Wild Ferment SEL); 12 monocellular haploids from original ones and 12 recuperated diploids from the monocells. The ethanol percentage ranged from 1.7 to 6.2% and the unfermentable reducing sugars from 0.45 to 0.50g/100mL. The highest ethanol percentages were produced by the monocellular cultures URM-MH3 (4.8%) in 12 h and SEL-MH1 (6.2%) in 24 h, corresponding to the productivity values of 3.15 and 2.03 g.L-1.h-1, respectively. The recuperated diploids did not present detectable ethanol content by the method used. The results showed that depending on the physiological state of the yeasts, being in the diploid or monocellular haploid form, different percentages of ethanol could be produced.

  11. Identification of the constitutive ultradian oscillator of the circadian clock (ENOX1 in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Sara S. Dick

    2013-05-01

    Full Text Available A yeast (Saccharomyces cerevisiae deletion library was screened based on NADH fluorescence using a 384 well plate assay and robotics to identify a yeast isolate lacking the 24 min periodic cell surface oxidase. The oxidase was shown previously to be a candidate ultradian oscillator of the yeast’s biological clock. The cDNA was cloned from a yeast overexpression library and the encoded protein was expressed in bacteria and characterized. Glyceraldehyde-3-phosphate dehydrogenase activity was used as the cellular circadian indicator. The identified gene was YML117W which encodes a ca 126 kDa putative RNA-binding protein. The candidate ENOX1 activity from yeast had functional characteristics similar to those of other constitutive ENOX1 proteins of eukaryotes exhibiting oscillating activities with a temperature independent period length of 24 min phased by melatonin and low frequency electromagnetic fields and susceptible to inhibition by the ENOX1 inhibitor, simalikalactone D. The YML117W deletion mutant cells lacked the ENOX1 clock output present in wild type yeast. The findings identify YML117W as the ENOX1 of Saccharomyces cerevisiae and support its proposed function as an ultradian oscillator of the yeast biological clock.

  12. Evaluation of cytochrome P-450 concentration in Saccharomyces cerevisiae strains

    Scientific Electronic Library Online (English)

    Míriam Cristina Sakuragui, Matuo; Irene Satiko, Kikuchi; Terezinha de Jesus Andreoli, Pinto.

    2010-09-01

    Full Text Available Linhagens de Saccharomyces cerevisiae tem sido amplamente empregadas em testes de mutagenicidade devido à presença de um sistema citocromo P-450 capaz de metabolizar substâncias pró-mutagênicas à sua forma ativa. Devido à grande variedade de linhagens de S. cerevisiae com diferentes capacidades de p [...] rodução de citocromo P-450, torna-se necessária a seleção de cepas, bem como a definição das condições ideais de cultivo. Neste trabalho, foram comparados os níveis de citocromo P-450 em quatro diferentes linhagens de S. cerevisiae e avaliadas as condições de cultivo necessárias para obtenção de altas concentrações deste sistema enzimático. O maior nível enzimático foi encontrado na linhagem NCYC 240 em presença de 20 % de glicose (p/v). A adição de etanol ao meio de cultura também produziu um aumento na síntese de citocromo P-450. Estes resultados indicam que as condições de cultivo devem ser específicas e bem definidas para a linhagem selecionada, garantindo assim elevados níveis de citocromo P-450 e, conseqüentemente, a confiabilidade nos testes de mutagenicidade. Abstract in english Saccharomyces cerevisiae has been widely used in mutagenicity tests due to the presence of a cytochrome P-450 system, capable of metabolizing promutagens to active mutagens. There are a large number of S. cerevisiae strains with varying abilities to produce cytochrome P-450. However, strain selectio [...] n and ideal cultivation conditions are not well defined. We compared cytochrome P-450 levels in four different S. cerevisiae strains and evaluated the cultivation conditions necessary to obtain the highest levels. The amount of cytochrome P-450 produced by each strain varied, as did the incubation time needed to reach the maximum level. The highest cytochrome P-450 concentrations were found in media containing fermentable sugars. The NCYC 240 strain produced the highest level of cytochrome P-450 when grown in the presence of 20 % (w/v) glucose. The addition of ethanol to the media also increased cytochrome P-450 synthesis in this strain. These results indicate cultivation conditions must be specific and well-established for the strain selected in order to assure high cytochrome P-450 levels and reliable mutagenicity results.

  13. Acquisition of tolerance against oxidative damage in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Eleutherio Elis CA

    2001-07-01

    Full Text Available Abstract Background Living cells constantly sense and adapt to redox shifts by the induction of genes whose products act to maintain the cellular redox environment. In the eukaryote Saccharomyces cerevisiae, while stationary cells possess a degree of constitutive resistance towards oxidants, treatment of exponential phase cultures with sub-lethal stresses can lead to the transient induction of protection against subsequent lethal oxidant conditions. The sensors of oxidative stress and the corresponding transcription factors that activate gene expression under these conditions have not yet been completely identified. Results We report the role of SOD1, SOD2 and TPS1 genes (which encode the cytoplasmic Cu/Zn-superoxide dismutase, the mitochondrial Mn-isoform and trehalose-6-phosphate synthase, respectively in the development of resistance to oxidative stress. In all experimental conditions, the cultures were divided into two parts, one was immediately submitted to severe stress (namely: exposure to H2O2, heat shock or ethanol stress while the other was initially adapted to 40°C for 60 min. The deficiency in trehalose synthesis did not impair the acquisition of tolerance to H2O2, but this disaccharide played an essential role in tolerance against heat and ethanol stresses. We also verified that the presence of only one Sodp isoform was sufficient to improve cellular resistance to 5 mM H2O2. On the other hand, while the lack of Sod2p caused high cell sensitivity to ethanol and heat shock, the absence of Sod1p seemed to be beneficial to the process of acquisition of tolerance to these adverse conditions. The increase in oxidation-dependent fluorescence of crude extracts of sod1 mutant cells upon incubation at 40°C was approximately 2-fold higher than in sod2 and control strain extracts. Furthermore, in Western blots, we observed that sod mutants showed a different pattern of Hsp104p and Hsp26p expression also different from that in their control strain. Conclusions Trehalose seemed not to be essential in the acquisition of tolerance to H2O2 stress, but its absence was strongly felt under water stress conditions such as heat and alcoholic stresses. On the other hand, Sod1p could be involved in the control of ROS production; these reactive molecules could signal the induction of genes implicated within cell tolerance to heat and ethanol. The effects of this deletion needs further investigation.

  14. Metabolic Engineering of Saccharomyces cerevisiae for Conversion of d-Glucose to Xylitol and Other Five-Carbon Sugars and Sugar Alcohols?

    OpenAIRE

    Toivari, Mervi H.; Ruohonen, Laura; Miasnikov, Andrei N.; Richard, Peter; Penttilä, Merja

    2007-01-01

    Recombinant Saccharomyces cerevisiae strains that produce the sugar alcohols xylitol and ribitol and the pentose sugar d-ribose from d-glucose in a single fermentation step are described. A transketolase-deficient S. cerevisiae strain accumulated d-xylulose 5-phosphate intracellularly and released ribitol and pentose sugars (d-ribose, d-ribulose, and d-xylulose) into the growth medium. Expression of the xylitol dehydrogenase-encoding gene XYL2 of Pichia stipitis in the transketolase-deficient...

  15. On the fermentative behavior of auxotrophic strains of Saccharomyces cerevisiae

    Scientific Electronic Library Online (English)

    Lucia, Paciello; Jesus, Zueco; Carmine, Landi.

    2014-09-15

    Full Text Available Background The selection of new yeast strains could lead to improvements in bioethanol production. Here, we have studied the fermentative capacity of different auxotrophic mutants of Saccharomyces cerevisiae, which are routinely used as hosts for the production of heterologous proteins. It has recen [...] tly been found that these strains exhibit physiological alterations and peculiar sensitivities with respect to the parental prototrophic strains from which they derive. In this work the performance of auxotrophic S. cerevisiae CEN.PK strains was compared to the corresponding prototrophic strain, to S. cerevisiae T5bV, a strain isolated from grape must and to another auxotrophic strain, S. cerevisiae BY4741. Results The results indicate that the fermentative capacity of strains grown in 2% glucose was similar in all the strains tested. However, in 15% initial glucose, the auxotrophic strains exhibited a more than doubled ethanol yield on biomass (10 g g- 1dw) compared to the prototrophic strains (less than 5 g g- 1dw). Other tests have also evidenced that in medium depletion conditions, ethanol production continues after growth arrest. Conclusions The results highlight the capacity of auxotrophic yeast strains to produce ethanol per mass unit, in a higher amount with respect to the prototrophic ones. This leads to potential applications for auxotrophic strains of S. cerevisiae in the production of ethanol in both homogeneous and heterogeneous phases (immobilized systems). The higher ethanol yield on biomass would be advantageous in immobilized cell systems, as a reduced yeast biomass could greatly reduce the mass transfer limitations through the immobilization matrix.

  16. A vaccine grade of yeast Saccharomyces cerevisiae expressing mammalian myostatin

    Directory of Open Access Journals (Sweden)

    Zhang Tingting

    2012-12-01

    Full Text Available Abstract Background Yeast Saccharomyces cerevisiae is a widely-used system for protein expression. We previously showed that heat-killed whole recombinant yeast vaccine expressing mammalian myostatin can modulate myostatin function in mice, resulting in increase of body weight and muscle composition in these animals. Foreign DNA introduced into yeast cells can be lost soon unless cells are continuously cultured in selection media, which usually contain antibiotics. For cost and safety concerns, it is essential to optimize conditions to produce quality food and pharmaceutical products. Results We developed a simple but effective method to engineer a yeast strain stably expressing mammalian myostatin. This method utilized high-copy-number integration of myostatin gene into the ribosomal DNA of Saccharomyces cerevisiae. In the final step, antibiotic selection marker was removed using the Cre-LoxP system to minimize any possible side-effects for animals. The resulting yeast strain can be maintained in rich culture media and stably express mammalian myostatin for two years. Oral administration of the recombinant yeast was able to induce immune response to myostatin and modulated the body weight of mice. Conclusions Establishment of such yeast strain is a step further toward transformation of yeast cells into edible vaccine to improve meat production in farm animals and treat human muscle-wasting diseases in the future.

  17. Engineering cellular redox balance in Saccharomyces cerevisiae for improved production of L-lactic acid.

    Science.gov (United States)

    Lee, Ju Young; Kang, Chang Duk; Lee, Seung Hyun; Park, Young Kyoung; Cho, Kwang Myung

    2015-04-01

    Owing to the growing market for the biodegradable and renewable polymer, polylactic acid, world demand for lactic acid is rapidly increasing. However, the very high concentrations desired for industrial production of the free lactic acid create toxicity and low pH concerns for manufacturers. Saccharomyces cerevisiae is the most well characterized eukaryote, a preferred microbial cell factory for the largest industrial biotechnology product (bioethanol), and a robust, commercially compatible workhorse to be exploited for the production of diverse chemicals. S. cerevisiae has also been explored as a host for lactic acid production because of its high acid tolerance. Here, we constructed an L-lactic acid-overproducing S. cerevisiae by redirecting cellular metabolic fluxes to the production of L-lactic acid. To this end, we deleted the S. cerevisiae genes encoding pyruvate decarboxylase 1 (PDC1), L-lactate cytochrome-c oxidoreductase (CYB2), and glycerol-3-phosphate dehydrogenase (GPD1), replacing them with a heterologous L-lactate dehydrogenase (LDH) gene. Two new target genes encoding isoenzymes of the external NADH dehydrogenase (NDE1 and NDE2), were also deleted from the genome to re-engineer the intracellular redox balance. The resulting strain was found to produce L-lactic acid more efficiently (32.6% increase in final L-lactic acid titer). When tested in a bioreactor in fed-batch mode, this engineered strain produced 117?g/L of L-lactic acid under low pH conditions. This result demonstrates that the redox balance engineering should be coupled with the metabolic engineering in the construction of L-lactic acid-overproducing S. cerevisiae. PMID:25363674

  18. Characterization of fungal RTG2 genes in retrograde signaling of Saccharomyces cerevisiae

    Science.gov (United States)

    Ünlü, Ercan Selçuk; Narayanan, Lakshmi; Gordon, Donna M

    2013-01-01

    Changes in the functional status of mitochondria result in the transcriptional activation of a subset of nuclear-encoded genes in a process referred to as retrograde signaling. In Saccharomyces cerevisiae, this molecular link between mitochondria and the nuclear genome is controlled by three key signaling proteins: Rtg1p, Rtg2p, and Rtg3p. Although the retrograde signaling response has been well characterized in S. cerevisiae, very little is known about this pathway in other fungi. In this study, we selected four species having uncharacterized open reading frames (ORFs) with more than 66% amino acid identity to Rtg2p for further analysis. To determine whether these putative RTG2 ORFs encoded bona fide regulators of retrograde signaling, we tested their ability to complement the defects associated with the S. cerevisiae rtg2? mutant. Specifically, we tested for complementation of citrate synthase (CIT2) and aconitase (ACO1) at the transcript and protein levels, glutamate auxotrophy, and changes in the interaction between Rtg2p and the negative regulator Mks1p. Our findings show that all four Rtg2p homologs are functional upon activation of retrograde signaling, although their degree of complementation varied. In addition, all Rtg2p homologs showed a marked reduction in Mks1p binding, which may contribute to their altered responses to retrograde signaling. PMID:23711018

  19. Novel physiological roles for glutathione in sequestering acetaldehyde to confer acetaldehyde tolerance in Saccharomyces cerevisiae.

    Science.gov (United States)

    Matsufuji, Yoshimi; Yamamoto, Kohei; Yamauchi, Kosei; Mitsunaga, Tohru; Hayakawa, Takashi; Nakagawa, Tomoyuki

    2013-01-01

    In this work, we identified novel physiological functions of glutathione in acetaldehyde tolerance in Saccharomyces cerevisiae. Strains deleted in the genes encoding the enzymes involved in glutathione synthesis and reduction, GSH1, GSH2 and GLR1, exhibited severe growth defects compared to wild-type under acetaldehyde stress, although strains deleted in the genes encoding glutathione peroxidases or glutathione transferases did not show any growth defects. On the other hand, intracellular levels of reduced glutathione decreased in the presence of acetaldehyde in response to acetaldehyde concentration. Moreover, we show that glutathione can trap a maximum of four acetaldehyde molecules within its molecule in a non-enzymatic manner. Taken together, these findings suggest that glutathione has an important role in acetaldehyde tolerance, as a direct scavenger of acetaldehyde in the cell. PMID:22615054

  20. Uranium removal from acidic aqueous solutions by Saccharomyces cerevisiae, Debaryomyces hansenii, Kluyveromyces marxianus and Candida colliculosa

    International Nuclear Information System (INIS)

    The sorption of uranium from acidic aqueous solutions (pH 4.5, Cinit = 10 to 1000 mg U/L) by Saccharomyces cerevisiae, Debaryomyces hansenii, Kluyveromyces marxianus and Candida colliculosa was investigated using a batch technique. The U-sorption onto Saccharomyces cerevisiae and Debaryomyces hansenii followed a Langmuir, while that onto Kluyveromyces marxianus and Candida colliculosa a Freundlich isotherm. The results demonstrated that all investigated biomasses could effectively remove uranium from acidic aqueous solutions. From all sorbents, Saccharomyces cerevisiae appeared to be the most effective with a maximum sorption capacity of 127.7 mg U/g dry biomass. (author)

  1. Increasing NADH oxidation reduces overflow metabolism in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Vemuri, Goutham; Eiteman, M.A

    2007-01-01

    Respiratory metabolism plays an important role in energy production in the form of ATP in all aerobically growing cells. However, a limitation in respiratory capacity results in overflow metabolism, leading to the formation of byproducts, a phenomenon known as ‘‘overflow metabolism’’ or ‘‘the Crabtree effect.’’ The yeast Saccharomyces cerevisiae has served as an important model organism for studying the Crabtree effect. When subjected to increasing glycolytic fluxes under aerobic conditions, there is a threshold value of the glucose uptake rate at which the metabolism shifts from purely respiratory to mixed respiratory and fermentative. It is well known that glucose repression of respiratory pathways occurs at high glycolytic fluxes, resulting in a decrease in respiratory capacity. Despite many years of detailed studies on this subject, it is not known whether the onset of the Crabtree effect is due to limited respiratory capacity or is caused by glucose-mediated repression of respiration. When respiration inS. cerevisiae was increased by introducing a heterologous alternative oxidase, we observed reduced aerobic ethanol formation. In contrast, increasing nonrespiratory NADH oxidation by overexpression of a water-forming NADH oxidase reduced aerobic glycerol formation. The metabolic response to elevated alternative oxidase occurred predominantly in the mitochondria, whereas NADH oxidase affected genes that catalyze cytosolic reactions. Moreover, NADH oxidase restored the deficiency of cytosolic NADH dehydrogenases in S. cerevisiae. These results indicate that NADH oxidase localizes in the cytosol, whereas alternative oxidase is directed to the mitochondria.

  2. Protein Enrichment of Cassava Pulp Fermentation by Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    C. Yuangklang

    2011-01-01

    Full Text Available The purpose of this study was to determine intestinal digestibility of residual components of cassava pulp solid state fermentation by Saccharomyces cerevisiae for animal feed. Three ruminally cannulated animal were used to measure in situ rumen Dry Matter (DM and Crude Protein (CP degradability characteristics of cassava pulp solid state fermentation by S. cerevisiae. Nylon bags containing 3 g (as fed basis of each feed was immersed in duplicate at each time point in the ventral rumen of each goat for 2, 4, 8, 12, 24, 48 and 72 h. Rumen feed residues from bags of 16 h incubation were used for estimation of lower gut digestibility by the technique of in vitro pepsin-pancreatin digestion. The results of the chemical analysis indicated that fermentation was slightly improved Ruminal Undegradable Protein (RUP of cassava pulp. The highest value of RUP was significantly differ (pS. cerevisiae in cassava pulp. The present results indicate that fermented cassava pulp can improve protein content and ruminal undegradable protein content.

  3. Irradiation effects on the alcohol fermentation ability of saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Irradiation effects on the alcohol fermentation ability of saccharomyces cerevisiae. S. cerevisiae suspensions of 1.5x108 clls/ml were exposed to single and fractionated doses of gamma irradiation, i.e. 0; 0.30; 0.60; 0.90; and 1.20 kGy in aerobic condition at dose rate of 1.63 kGy/hour. The fractionated doses were given with time interval of 15, 30 and 45 minutes. The fermentation was held at 300C for 40 hours. It is seen that an increase of alcohol production was obtained when cells were irradiated at 0.60 kGy, although the result has no significant difference statistically with control. At the dose of 1.20 kGy the alcohol fermentation ability of S. cerevisiae decreased drastically as compared to control. Irradiation using single or fractionated doses with time interval of 15-45 minutes did not influence the alcohol production. Comparing the time interval of 45 minutes at 0.60 kGy and at 1.20 kGy, it appeared that the yield of alcohol was different. (author). 17 refs.; 4 figs

  4. Osmo-, thermo- and ethanol- tolerances of Saccharomyces cerevisiae S1

    Scientific Electronic Library Online (English)

    Sandrasegarampillai, Balakumar; Vasanthy, Arasaratnam.

    2012-03-01

    Full Text Available Saccharomyces cerevisiae S1, which is a locally isolated and improved strain showed viability at 40, 45 and 50ºC and produced ethanol at 40, 43 and 45ºC. When the cells were given heat shock at 45ºC for 30min and grown at 40ºC, 100% viability was observed for 60h, and addition of 200gl-1 ethanol has [...] led to complete cell death at 30h. Heat shock given at 45ºC (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-1 sorbitol. In presence of sorbitol (200gl-1) and ethanol (50gl-1) at 40ºC, 46% viability was retained by S. cerevisiae S1 at 48h and it was improved to 80% by soy flour supplementation.

  5. Capacidade fermentativa de Saccharomyces cerevisiae enriquecida com ácidos graxos / Fermentative capacity of (Saccharomyces cerevisiae) enriched with fatty acids

    Scientific Electronic Library Online (English)

    L.E., Gutierrez; A.V.K.O., Annicchino; L., Lucatti.

    Full Text Available Levedura de panificaçao (Saccharomyces cerevisiae) foi obtida anaerobicamente (sem e com adição dos ácidos graxos palmítico, oleico e linoleico) e aerobicamente e utilizada em ensaios de fermentação com 14% e 16% de sacarose a 32°C. Não houve diferenças significativas, quanto a viabilidade celular, [...] entre os tratamentos das leveduras com ácido oleico, ácido linoleico e aerobicamente (as quais foram ricas em palmitoleico e oleico). As leveduras enriquecidas com ácido palmítico e anaeróbicas apresentaram maior redução na viabilidade do que com ácidos graxos insaturados. Foi observado um aumento na produção de ácido pirúvico e uma redução nos álcoois superiores com a redução da viabilidade celular. Abstract in english Baker's yeast (Saccharomyces cerevisiae) was obtained anaerobically (with and without the addition of following fatty acids: palmitic, oleic and linoleic) aerobically and utilised infermentation trials with 14 and 16% of sucrose in the medium at 32°C. There were no significant differences among olei [...] c acid, linoleic acid and aerobic treatments (which were rich in palmitoleic and oleic acids) in relation to cellular viability. Yeasts enriched with palmitic acid and under anaerobic conditions showed a higher reduction on viability than those treated with unsaturated fatty acids. An increased production of pyruvic acid and a reduction in higher alcohols with a reduction on cellular viability were observed.

  6. Comparison of heterologous xylose transporters in recombinant Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Hahn-Hägerdal Bärbel

    2010-03-01

    Full Text Available Abstract Background Baker's yeast (Saccharomyces cerevisiae has been engineered for xylose utilization to enable production of fuel ethanol from lignocellulose raw material. One unresolved challenge is that S. cerevisiae lacks a dedicated transport system for pentose sugars, which means that xylose is transported by non-specific Hxt transporters with comparatively low transport rate and affinity for xylose. Results In this study, we compared three heterologous xylose transporters that have recently been shown to improve xylose uptake under different experimental conditions. The transporters Gxf1, Sut1 and At5g59250 from Candida intermedia, Pichia stipitis and Arabidopsis thaliana, respectively, were expressed in isogenic strains of S. cerevisiae and the transport kinetics and utilization of xylose was evaluated. Expression of the Gxf1 and Sut1 transporters led to significantly increased affinity and transport rates of xylose. In batch cultivation at 4 g/L xylose concentration, improved transport kinetics led to a corresponding increase in xylose utilization, whereas no correlation could be demonstrated at xylose concentrations greater than 15 g/L. The relative contribution of native sugar transporters to the overall xylose transport capacity was also estimated during growth on glucose and xylose. Conclusions Kinetic characterization and aerobic batch cultivation of strains expressing the Gxf1, Sut1 and At5g59250 transporters showed a direct relationship between transport kinetics and xylose growth. The Gxf1 transporter had the highest transport capacity and the highest xylose growth rate, followed by the Sut1 transporter. The range in which transport controlled the growth rate was determined to between 0 and 15 g/L xylose. The role of catabolite repression in regulation of native transporters was also confirmed by the observation that xylose transport by native S. cerevisiae transporters increased significantly during cultivation in xylose and at low glucose concentration.

  7. Heterologous production of non-ribosomal peptide LLD-ACV in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Siewers, Verena; Chen, Xiao

    2009-01-01

    Non-ribosomal peptides (NRPs) are a diverse family of secondary metabolites with a broad range of biological activities. We started to develop an eukaryotic microbial platform based on the yeast Saccharomyces cerevisiae for heterologous production of NRPs using ?-(l-?-aminoadipyl)–l-cysteinyl–d-valine (ACV) as a model NRP. The Penicillium chrysogenum gene pcbAB encoding ACV synthetase was expressed in S. cerevisiae from a high-copy plasmid together with phosphopantetheinyl transferase (PPTase) encoding genes from Aspergillus nidulans, P. chrysogenum and Bacillus subtilis, and in all the three cases production of ACV was observed. To improve ACV synthesis, several factors were investigated. Codon optimization of the 5? end of pcbAB did not significantly increase ACV production. However, a 30-fold enhancement was achieved by lowering the cultivation temperature from 30 to 20 °C. When ACVS and PPTase encoding genes were integrated into the yeast genome, a 6-fold decrease in ACV production was observed indicatingthat gene copy number was one of the rate-limiting factors for ACV production in yeast.

  8. “A comparison between sugar consumption and ethanol production in wort by immobilized Saccharomyces Cerevisiae, Saccharomyces Ludwigii and Saccharomyces Rouxii on Brewer’S Spent Grain”

    OpenAIRE

    Mohammadi, Aniseh; Razavi, Seyyed Hadi; Mousavi, Seyyed Mohammad; Rezaei, Karamatollah

    2011-01-01

    The immobilization of Saccharomyces cerevisiae DSM 70424, Saccharomyces ludwigii DSM 3447 and Saccharomyces rouxii DSM 2531 on brewer’s spent grain and then ethanol production and sugar consumption of these immobilized yeasts were investigated. The aim of this study was to investigate the abilities of these three immobilized yeasts for producing alcohol for brewing at two temperatures (7 and 12 °C) using two different sugar levels (one at original level supplied in the brewery and one with...

  9. RPG1úTIF32 Saccharomyces cerevisiae a jeho interakce s miktotubuly.

    Czech Academy of Sciences Publication Activity Database

    Špryngar, Martin; Janatová, Ivana; Hašek, Ji?í

    Praha, 2002. s. -. [Cytoskeletální klub /9./. 25.04.2001-27.04.2001, Vranovská Ves] R&D Projects: GA ?R GA204/99/1531 Keywords : rpg * saccharomyces * cerevisiae Subject RIV: EE - Microbiology, Virology

  10. Probing glycolytic and membrane potential oscillations in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Poulsen, Allan K.; Andersen, Ann Zahle

    2008-01-01

    We have investigated glycolytic oscillations under semi-anaerobic conditions in Saccharomyces cerevisiae by means of NADH fluorescence, measurements of intracellular glucose concentration, and mitochondrial membrane potential. The glucose concentration was measured using an optical nanosensor, while mitochondrial membrane potential was measured using the fluorescent dye DiOC(2)(3). The results show that, as opposed to NADH and other intermediates in glycolysis, intracellular glucose is not oscillating. Furthermore, oscillations in NADH and membrane potential are inhibited by the ATP/ADP antiporter inhibitor atractyloside and high concentrations of the ATPase inhibitor N,N'-dicyclohexylcarbodiimide, suggesting that there is a strong coupling between oscillations in mitochondrial membrane potential and oscillations in NADH mediated by the ATP/ADP antiporter and possibly also other respiratory components.

  11. Replication of Avocado Sunblotch Viroid in the Yeast Saccharomyces cerevisiae?

    Science.gov (United States)

    Delan-Forino, Clémentine; Maurel, Marie-Christine; Torchet, Claire

    2011-01-01

    Viroids are the smallest known pathogenic agents. They are noncoding, single-stranded, closed-circular, “naked” RNAs, which replicate through RNA-RNA transcription. Viroids of the Avsunviroidae family possess a hammerhead ribozyme in their sequence, allowing self-cleavage during their replication. To date, viroids have only been detected in plant cells. Here, we investigate the replication of Avocado sunblotch viroid (ASBVd) of the Avsunviroidae family in a nonconventional host, the yeast Saccharomyces cerevisiae. We demonstrate that ASBVd RNA strands of both polarities are able to self-cleave and to replicate in a unicellular eukaryote cell. We show that the viroid monomeric RNA is destabilized by the nuclear 3? and the cytoplasmic 5? RNA degradation pathways. For the first time, our results provide evidence that viroids can replicate in other organisms than plants and that yeast contains all of the essential cellular elements for the replication of ASBVd. PMID:21270165

  12. Tolerance of budding yeast Saccharomyces cerevisiae to ultra high pressure

    Science.gov (United States)

    Shibata, M.; Torigoe, M.; Matsumoto, Y.; Yamamoto, M.; Takizawa, N.; Hada, Y.; Mori, Y.; Takarabe, K.; Ono, F.

    2014-05-01

    Our studies on the tolerance of plants and animals against very high pressure of several GPa have been extended to a smaller sized fungus, the budding yeast Saccharomyces cerevisiae. Several pieces of budding yeast (dry yeast) were sealed in a small teflon capsule with a liquid pressure medium fluorinate, and exposed to 7.5 GPa by using a cubic anvil press. The pressure was kept constant for various duration of time from 2 to 24 h. After the pressure was released, the specimens were brought out from the teflon capsule, and they were cultivated on a potato dextrose agar. It was found that the budding yeast exposed to 7.5 GPa for up to 6 h showed multiplication. However, those exposed to 7.5 GPa for longer than 12 h were found dead. The high pressure tolerance of budding yeast is a little weaker than that of tardigrades.

  13. Higher-order structure of Saccharomyces cerevisiae chromatin

    International Nuclear Information System (INIS)

    We have developed a method for partially purifying chromatin from Saccharomyces cerevisiae (baker's yeast) to a level suitable for studies of its higher-order folding. This has required the use of yeast strains that are free of the ubiquitous yeast killer virus. Results from dynamic light scattering, electron microscopy, and x-ray diffraction show that the yeast chromatin undergoes a cation-dependent folding into 30-nm filaments that resemble those characteristic of higher-cell chromatin; moreover, the packing of nucleosomes within the yeast 30-nm filaments is similar to that of higher cells. These results imply that yeast has a protein or protein domain that serves the role of the histone H 1 found in higher cells; physical and genetic studies of the yeast activity could help elucidate the structure and function of H 1. Images of the yeast 30-nm filaments can be used to test crossed-linker models for 30-nm filament structure

  14. Adaptive answer to low ionizing radiation doses in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    The aim of this work is to verify the existence of the adaptive response phenomenon induced by low doses of ionizing radiation in living cells. It is known that low doses of ionizing radiation, called conditioning doses, may induce resistance in exposed organisms to higher doses, called challenging doses, which are applied after a period of time. The involved mechanisms in this phenomenon, called Adaptive Response, are diverse and complex. Among them, the most important are the activation of DNA-repair enzymes and nuclear recombination process. As the 'target' sample, it was utilized a 'wild type' strain of Saccharomyces cerevisiae in aqueous suspension. Adaptive Response was verified in a wide range of challenging doses. Conditioning doses, inductors of radio-resistance, were (0.44±0.03) Gy and the waiting time between them and challenging doses was 2 hours at room temperature.(author)

  15. Brazilian propolis protects Saccharomyces cerevisiae cells against oxidative stress

    Scientific Electronic Library Online (English)

    Rafael A. de, Sá; Frederico A.V. de, Castro; Elis C.A., Eleutherio; Raquel M. de, Souza; Joaquim F.M. da, Silva; Marcos D., Pereira.

    2013-09-01

    Full Text Available Propolis is a natural product widely used for humans. Due to its complex composition, a number of applications (antimicrobial, antiinflammatory, anesthetic, cytostatic and antioxidant) have been attributed to this substance. Using Saccharomyces cerevisiae as a eukaryotic model we investigated the me [...] chanisms underlying the antioxidant effect of propolis from Guarapari against oxidative stress. Submitting a wild type (BY4741) and antioxidant deficient strains (ctt1?, sod1?, gsh1?, gtt1? and gtt2?) either to 15 mM menadione or to 2 mM hydrogen peroxide during 60 min, we observed that all strains, except the mutant sod1?, acquired tolerance when previously treated with 25 µg/mL of alcoholic propolis extract. Such a treatment reduced the levels of ROS generation and of lipid peroxidation, after oxidative stress. The increase in Cu/Zn-Sod activity by propolis suggests that the protection might be acting synergistically with Cu/Zn-Sod.

  16. ACTIVITY OF SUPEROXIDE DISMUTASE ENZYME IN YEAST SACCHAROMYCES CEREVISIAE

    Directory of Open Access Journals (Sweden)

    Blažena Lavová

    2014-02-01

    Full Text Available Reactive oxygen species (ROS with reactive nitrogen species (RNS are known to play dual role in biological systems, they can be harmful or beneficial to living systems. ROS can be important mediators of damage to cell structures, including proteins, lipids and nucleic acids termed as oxidative stress. The antioxidant enzymes protect the organism against the oxidative damage caused by active oxygen forms. The role of superoxide dismutase (SOD is to accelerate the dismutation of the toxic superoxide radical, produced during oxidative energy processes, to hydrogen peroxide and molecular oxygen. In this study, SOD activity of three yeast strains Saccharomyces cerevisiae was determined. It was found that SOD activity was the highest (23.7 U.mg-1 protein in strain 612 after 28 hours of cultivation. The lowest SOD activity from all tested strains was found after 56 hours of cultivation of strain Gyöng (0.7 U.mg-1 protein.

  17. Pyrimidine-specific cleavage by an endoribonuclease of Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    An endoribonuclease with pyrimidine cleavage site specificity was isolated from Saccharomyces cerevisiae. The enzyme had a pH optimum of 6 to 7 and did not require a divalent cation. It was inhibited by 5 x 10-5 M hetidium bromide, although it appeared to be single strand specific. The enzyme gave a limited cleavage of yeast mRNA and rRNA, yielding products that were terminated with pyrimidine nucleoside 2',3'-cyclic phosphate. The bonds between pyrimidine and A residues constituted more than 90% of the scission sites when the average product size was 50 nucloetides. Homopolyribonucleotides were cleaved poorly. Poly(A,U) was cleaved rapidly, and analysis of the products of poly(A,U) hydrolysis showed a very stringent cleavage of U-A bonds

  18. Structural properties of Saccharomyces cerevisiae protein complex network

    CERN Document Server

    Ramezanpour, A; Karimipour, V

    2003-01-01

    Recent studies indicate that protein complexes rather than individual proteins are the functional units of the cell. A first step in the understanding of integration and coordination of these cellular functions is a comprehensive study of the structural features of the network of connections between these complexes. We use the recently obtained data based on tandem-affinity purification (TAP) and mass spectrometry, to study the structural features of the yeast Saccharomyces cerevisiae protein complex network. We find striking similarities and differences between the structural properties of the networks of proteins and protein complexes. The network of complexes is still a small world network with scale free distributions for many of its properties (e.g. connectivity and size of complexes). However in contrast to the protein network, we find no correlations between the degrees of neighbouring complexes. There is also no correlation between the size of neighbouring complexes. Finally we propose a simple evolut...

  19. Effect of Reserve Carbohydrates on Oxidative Stress in Yeast Saccharomyces cerevisiae Y6210

    OpenAIRE

    Smita Kanwal; Saharan, R. K.; Mahmood, A.; Sharma, S. C.

    2011-01-01

    The aim of this study is investigate the role of reserve carbohydrates, trehalose and glycogen in DEM (Diethyl Maleate) induced oxidative stress in yeast Saccharomyces cerevisiae Y6210. Trehalose and glycogen accumulated in Saccharomyces cerevisiae, when growth conditions deteriorate. Yeast cells were subjected to oxidative stress for different time periods (0, 30, 60 and 120 min) to evaluate the role of trehalose, glycogen and trehalase. There was no change in the level of trehalose while th...

  20. Repair of UV-damaged incoming plasmid DNA in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    A whole-cell transformation assay was used for the repair of UV-damaged plasma DNA in highly-transformable haploid strains of Saccharomyces cerevisiae having different repair capabilities. The experiments described demonstrate that three epistasis groups (Friedberg 1988) are involved in the repair of UV-incoming DNA and that the repair processes act less efficiently on incoming DNA than they do on chromosomal DNA. The implications of these findings for UV repair in Saccharomyces cerevisiae are discussed. (author)

  1. Water treatment process and system for metals removal using Saccharomyces cerevisiae

    Science.gov (United States)

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

    2002-01-01

    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.

  2. Copper Tolerance and Biosorption of Saccharomyces cerevisiae during Alcoholic Fermentation.

    Science.gov (United States)

    Sun, Xiang-Yu; Zhao, Yu; Liu, Ling-Ling; Jia, Bo; Zhao, Fang; Huang, Wei-Dong; Zhan, Ji-Cheng

    2015-01-01

    At high levels, copper in grape mash can inhibit yeast activity and cause stuck fermentations. Wine yeast has limited tolerance of copper and can reduce copper levels in wine during fermentation. This study aimed to understand copper tolerance of wine yeast and establish the mechanism by which yeast decreases copper in the must during fermentation. Three strains of Saccharomyces cerevisiae (lab selected strain BH8 and industrial strains AWRI R2 and Freddo) and a simple model fermentation system containing 0 to 1.50 mM Cu2+ were used. ICP-AES determined Cu ion concentration in the must decreasing differently by strains and initial copper levels during fermentation. Fermentation performance was heavily inhibited under copper stress, paralleled a decrease in viable cell numbers. Strain BH8 showed higher copper-tolerance than strain AWRI R2 and higher adsorption than Freddo. Yeast cell surface depression and intracellular structure deformation after copper treatment were observed by scanning electron microscopy and transmission electron microscopy; electronic differential system detected higher surface Cu and no intracellular Cu on 1.50 mM copper treated yeast cells. It is most probably that surface adsorption dominated the biosorption process of Cu2+ for strain BH8, with saturation being accomplished in 24 h. This study demonstrated that Saccharomyces cerevisiae strain BH8 has good tolerance and adsorption of Cu, and reduces Cu2+ concentrations during fermentation in simple model system mainly through surface adsorption. The results indicate that the strain selected from China's stress-tolerant wine grape is copper tolerant and can reduce copper in must when fermenting in a copper rich simple model system, and provided information for studies on mechanisms of heavy metal stress. PMID:26030864

  3. Global Mapping of DNA Conformational Flexibility on Saccharomyces cerevisiae.

    Science.gov (United States)

    Menconi, Giulia; Bedini, Andrea; Barale, Roberto; Sbrana, Isabella

    2015-04-01

    In this study we provide the first comprehensive map of DNA conformational flexibility in Saccharomyces cerevisiae complete genome. Flexibility plays a key role in DNA supercoiling and DNA/protein binding, regulating DNA transcription, replication or repair. Specific interest in flexibility analysis concerns its relationship with human genome instability. Enrichment in flexible sequences has been detected in unstable regions of human genome defined fragile sites, where genes map and carry frequent deletions and rearrangements in cancer. Flexible sequences have been suggested to be the determinants of fragile gene proneness to breakage; however, their actual role and properties remain elusive. Our in silico analysis carried out genome-wide via the StabFlex algorithm, shows the conserved presence of highly flexible regions in budding yeast genome as well as in genomes of other Saccharomyces sensu stricto species. Flexibile peaks in S. cerevisiae identify 175 ORFs mapping on their 3'UTR, a region affecting mRNA translation, localization and stability. (TA)n repeats of different extension shape the central structure of peaks and co-localize with polyadenylation efficiency element (EE) signals. ORFs with flexible peaks share common features. Transcripts are characterized by decreased half-life: this is considered peculiar of genes involved in regulatory systems with high turnover; consistently, their function affects biological processes such as cell cycle regulation or stress response. Our findings support the functional importance of flexibility peaks, suggesting that the flexible sequence may be derived by an expansion of canonical TAYRTA polyadenylation efficiency element. The flexible (TA)n repeat amplification could be the outcome of an evolutionary neofunctionalization leading to a differential 3'-end processing and expression regulation in genes with peculiar function. Our study provides a new support to the functional role of flexibility in genomes and a strategy for its characterization inside human fragile sites. PMID:25860149

  4. Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol

    Directory of Open Access Journals (Sweden)

    Myers Samuel

    2008-12-01

    Full Text Available Abstract Background Increasing energy costs and environmental concerns have motivated engineering microbes for the production of "second generation" biofuels that have better properties than ethanol. Results and conclusion Saccharomyces cerevisiae was engineered with an n-butanol biosynthetic pathway, in which isozymes from a number of different organisms (S. cerevisiae, Escherichia coli, Clostridium beijerinckii, and Ralstonia eutropha were substituted for the Clostridial enzymes and their effect on n-butanol production was compared. By choosing the appropriate isozymes, we were able to improve production of n-butanol ten-fold to 2.5 mg/L. The most productive strains harbored the C. beijerinckii 3-hydroxybutyryl-CoA dehydrogenase, which uses NADH as a co-factor, rather than the R. eutropha isozyme, which uses NADPH, and the acetoacetyl-CoA transferase from S. cerevisiae or E. coli rather than that from R. eutropha. Surprisingly, expression of the genes encoding the butyryl-CoA dehydrogenase from C. beijerinckii (bcd and etfAB did not improve butanol production significantly as previously reported in E. coli. Using metabolite analysis, we were able to determine which steps in the n-butanol biosynthetic pathway were the most problematic and ripe for future improvement.

  5. Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol

    Energy Technology Data Exchange (ETDEWEB)

    Steen, EricJ.; Chan, Rossana; Prasad, Nilu; Myers, Samuel; Petzold, Christopher; Redding, Alyssa; Ouellet, Mario; Keasling, JayD.

    2008-11-25

    BackgroundIncreasing energy costs and environmental concerns have motivated engineering microbes for the production of ?second generation? biofuels that have better properties than ethanol.Results& ConclusionsSaccharomyces cerevisiae was engineered with an n-butanol biosynthetic pathway, in which isozymes from a number of different organisms (S. cerevisiae, Escherichia coli, Clostridium beijerinckii, and Ralstonia eutropha) were substituted for the Clostridial enzymes and their effect on n-butanol production was compared. By choosing the appropriate isozymes, we were able to improve production of n-butanol ten-fold to 2.5 mg/L. The most productive strains harbored the C. beijerinckii 3-hydroxybutyryl-CoA dehydrogenase, which uses NADH as a co-factor, rather than the R. eutropha isozyme, which uses NADPH, and the acetoacetyl-CoA transferase from S. cerevisiae or E. coli rather than that from R. eutropha. Surprisingly, expression of the genes encoding the butyryl-CoA dehydrogenase from C. beijerinckii (bcd and etfAB) did not improve butanol production significantly as previously reported in E. coli. Using metabolite analysis, we were able to determine which steps in the n-butanol biosynthetic pathway were the most problematic and ripe for future improvement.

  6. Genomic Evolution of Saccharomyces cerevisiae under Chinese Rice Wine Fermentation

    Science.gov (United States)

    Li, Yudong; Zhang, Weiping; Zheng, Daoqiong; Zhou, Zhan; Yu, Wenwen; Zhang, Lei; Feng, Lifang; Liang, Xinle; Guan, Wenjun; Zhou, Jingwen; Chen, Jian; Lin, Zhenguo

    2014-01-01

    Rice wine fermentation represents a unique environment for the evolution of the budding yeast, Saccharomyces cerevisiae. To understand how the selection pressure shaped the yeast genome and gene regulation, we determined the genome sequence and transcriptome of a S. cerevisiae strain YHJ7 isolated from Chinese rice wine (Huangjiu), a popular traditional alcoholic beverage in China. By comparing the genome of YHJ7 to the lab strain S288c, a Japanese sake strain K7, and a Chinese industrial bioethanol strain YJSH1, we identified many genomic sequence and structural variations in YHJ7, which are mainly located in subtelomeric regions, suggesting that these regions play an important role in genomic evolution between strains. In addition, our comparative transcriptome analysis between YHJ7 and S288c revealed a set of differentially expressed genes, including those involved in glucose transport (e.g., HXT2, HXT7) and oxidoredutase activity (e.g., AAD10, ADH7). Interestingly, many of these genomic and transcriptional variations are directly or indirectly associated with the adaptation of YHJ7 strain to its specific niches. Our molecular evolution analysis suggested that Japanese sake strains (K7/UC5) were derived from Chinese rice wine strains (YHJ7) at least approximately 2,300 years ago, providing the first molecular evidence elucidating the origin of Japanese sake strains. Our results depict interesting insights regarding the evolution of yeast during rice wine fermentation, and provided a valuable resource for genetic engineering to improve industrial wine-making strains. PMID:25212861

  7. Quantifying separation and similarity in a Saccharomyces cerevisiae metapopulation.

    Science.gov (United States)

    Knight, Sarah; Goddard, Matthew R

    2015-02-01

    Eukaryotic microbes are key ecosystem drivers; however, we have little theory and few data elucidating the processes influencing their observed population patterns. Here we provide an in-depth quantitative analysis of population separation and similarity in the yeast Saccharomyces cerevisiae with the aim of providing a more detailed account of the population processes occurring in microbes. Over 10,000 individual isolates were collected from native plants, vineyards and spontaneous ferments of fruit from six major regions spanning 1000?km across New Zealand. From these, hundreds of S. cerevisiae genotypes were obtained, and using a suite of analytical methods we provide comprehensive quantitative estimates for both population structure and rates of gene flow or migration. No genetic differentiation was detected within geographic regions, even between populations inhabiting native forests and vineyards. We do, however, reveal a picture of national population structure at scales above ?100?km with distinctive populations in the more remote Nelson and Central Otago regions primarily contributing to this. In addition, differential degrees of connectivity between regional populations are observed and correlate with the movement of fruit by the New Zealand wine industry. This suggests some anthropogenic influence on these observed population patterns. PMID:25062126

  8. Induction and inhibition of the allantoin permease in Saccharomyces cerevisiae.

    Science.gov (United States)

    Sumrada, R; Zacharski, C A; Turoscy, V; Cooper, T G

    1978-01-01

    Allantoin uptake in Saccharomyces cerevisiae is mediated by an energy-dependent, low-Km, active transport system. However, there is at present little information concerning its regulation. In view of this, we investigated the control of alloantoin transport and found that it was regulated quite differently from the other pathway components. Preincubation of appropriate mutant cultures with purified allantoate (commercial preparations contain 17% allantoin), urea, or oxalurate did not significantly increase allantoin uptake. Preincubation with allantoin, however, resulted in a 10- to 15-fold increase in the rate of allantoin accumulation. Two allantoin analogs were also found to elicit dramatic increases in allantoin uptake. Hydantoin and hydantoin acetic acid were able to induce allantoin transport to 63 and 95% of the levels observed with allantoin. Neither of these compounds was able to serve as a sole nitrogen source for S. cerevisiae, and they may be non-metabolizable inducers of the allantoin permease. The rna1 gene product appeared to be required for allantoin permease induction, suggesting that control was exerted at the level of gene expression. In addition, we have shown that allantoin uptake is not unidirectional; efflux merely occurs at a very low rate. Allantoin uptake is also transinhibited by addition of certain amino acids to the culture medium, and several models concerning the operation of such inhibition were discussed. PMID:355231

  9. Long-chain alkane production by the yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Buijs, Nicolaas A; Zhou, Yongjin J; Siewers, Verena; Nielsen, Jens

    2015-06-01

    In the past decade industrial-scale production of renewable transportation biofuels has been developed as an alternative to fossil fuels, with ethanol as the most prominent biofuel and yeast as the production organism of choice. However, ethanol is a less efficient substitute fuel for heavy-duty and maritime transportation as well as aviation due to its low energy density. Therefore, new types of biofuels, such as alkanes, are being developed that can be used as drop-in fuels and can substitute gasoline, diesel, and kerosene. Here, we describe for the first time the heterologous biosynthesis of long-chain alkanes by the yeast Saccharomyces cerevisiae. We show that elimination of the hexadecenal dehydrogenase Hfd1 and expression of a redox system are essential for alkane biosynthesis in yeast. Deletion of HFD1 together with expression of an alkane biosynthesis pathway resulted in the production of the alkanes tridecane, pentadecane, and heptadecane. Our study provides a proof of principle for producing long-chain alkanes in the industrial workhorse S. cerevisiae, which was so far limited to bacteria. We anticipate that these findings will be a key factor for further yeast engineering to enable industrial production of alkane based drop-in biofuels, which can allow the biofuel industry to diversify beyond bioethanol. Biotechnol. Bioeng. 2015;112: 1275-1279. © 2014 Wiley Periodicals, Inc. PMID:25545362

  10. Heat shock response improves heterologous protein secretion in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Hou, Jin; Österlund, Tobias

    2013-01-01

    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.

  11. High level secretion of cellobiohydrolases by Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Ahlgren Simon

    2011-09-01

    Full Text Available Abstract 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.

  12. Improvement of Galactose Uptake in Saccharomyces cerevisiae through Overexpression of Phosphoglucomutase: Example of Transcript Analysis as a Tool in Inverse Metabolic Engineering

    OpenAIRE

    Bro, Christoffer; Knudsen, Steen; Regenberg, Birgitte; Olsson, Lisbeth; Nielsen, Jens

    2005-01-01

    Through genome-wide transcript analysis of a reference strain and two recombinant Saccharomyces cerevisiae strains with different rates of galactose uptake, we obtained information about the global transcriptional response to metabolic engineering of the GAL gene regulatory network. One of the recombinant strains overexpressed the gene encoding the transcriptional activator Gal4, and in the other strain the genes encoding Gal80, Gal6, and Mig1, which are negative regulators of the GAL system,...

  13. Reconstruction of the biosynthetic pathway for the core fungal polyketide scaffold rubrofusarin in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Rugbjerg, Peter; Naesby, Michael

    2013-01-01

    ABSTRACT: BACKGROUND: Fungal polyketides include commercially important pharmaceuticals and food additives, e.g. the cholesterol-lowering statins and the red and orange monascus pigments. Presently, production relies on isolation of the compounds from the natural producers, and systems for heterologous production in easily fermentable and genetically engineerable organisms, such as Saccharomyces cerevisiae and Escherichia coli are desirable. Rubrofusarin is an orange polyketide pigment that is a common intermediate in many different fungal biosynthetic pathways. RESULTS: In this study, we established a biosynthetic pathway for rubrofusarin in S. cerevisiae. First, the Fusarium graminearum gene encoding polyketide synthase 12 (PKS12) was heterologously co-expressed with the Aspergillus fumigatus gene encoding phosphopantetheinyl transferase (npgA) resulting in production of YWA1. This aromatic heptaketide intermediate was converted into nor-rubrofusarin upon expression of the dehydratase gene aurZ from the aurofusarin gene cluster of F. graminearum. Final conversion into rubrofusarin was achieved by expression of the O-methyltransferase encoding gene aurJ, also obtained from the aurofusarin gene cluster, resulting in a titer of 1.1 mg/L. Reduced levels of rubrofusarin were detected when expressing PKS12, npgA, and aurJ alone, presumably due to spontaneous conversion of YWA1 to nor-rubrofusarin. However, the co-expression of aurZ resulted in an approx. six-fold increase in rubrofusarin production. CONCLUSIONS: The reconstructed pathway for rubrofusarin in S. cerevisiae allows the production of a core scaffold molecule with a branch-point role in several fungal polyketide pathways, thus paving the way for production of further natural pigments and bioactive molecules. Furthermore, the reconstruction verifies the suggested pathway, and as such, it is the first example of utilizing a synthetic biological “bottom up” approach for the validation of a complex fungal polyketide pathway.

  14. Invertase SUC2 Is the Key Hydrolase for Inulin Degradation in Saccharomyces cerevisiae

    OpenAIRE

    Wang, Shi-An; Li, Fu-li

    2013-01-01

    Specific Saccharomyces cerevisiae strains were recently found to be capable of efficiently utilizing inulin, but genetic mechanisms of inulin hydrolysis in yeast remain unknown. Here we report functional characteristics of invertase SUC2 from strain JZ1C and demonstrate that SUC2 is the key enzyme responsible for inulin metabolism in S. cerevisiae.

  15. ISOLATION OF A CYTOCHROME P-450 STRUCTURAL GENE FROM SACCHAROMYCES CEREVISIAE

    Science.gov (United States)

    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...

  16. Enhancing sesquiterpene production in Saccharomyces cerevisiae through in silico driven metabolic engineering

    DEFF Research Database (Denmark)

    Asadollahi, Mohammadali; Maury, Jerome

    2009-01-01

    A genome-scale metabolic model was used to identify new target genes for enhanced biosynthesis of sesquiterpenes in the yeast Saccharomyces cerevisiae. The effect of gene deletions on the flux distributions in the metabolic model of S. cerevisiae was assessed using OptGene as the modeling framework and minimization of metabolic adjustments (MOMA) as objective function. Deletion of NADPH-dependent glutamate dehydrogenase encoded by GDH1 was identified as the best target gene for the improvement of sesquiterpene biosynthesis in yeast. Deletion of this gene enhances the available NADPH in the cytosol for other NADPH requiring enzymes, including HMG-CoA reductase. However, since disruption of GDH1 impairs the ammonia utilization, simultaneous over-expression of the NADH-dependent glutamate dehydrogenase encoded by GDH2 was also considered in this study. Deletion of GDH1 led to an approximately 85% increase in the final cubebol titer. However, deletion of this gene also caused a significant decrease in the maximum specific growth rate. Over-expression of GDH2 did not show a further effect on the final cubebol titer but this alteration significantly improved the growth rate compared to the GDH1 deleted strain.

  17. Crystallization and preliminary X-ray diffraction analysis of the invertase from Saccharomyces cerevisiae.

    Science.gov (United States)

    Sainz-Polo, M Angela; Lafraya, Alvaro; Polo, Aitana; Marín-Navarro, Julia; Polaina, Julio; Sanz-Aparicio, Julia

    2012-12-01

    Saccharomyces cerevisiae invertase (ScInv) is an enzyme encoded by the SUC2 gene that releases ?-fructose from the nonreducing termini of various ?-D-fructofuranoside substrates. Its ability to produce 6-kestose by transglycosylation makes this enzyme an interesting research target for applications in industrial biotechnology. The native enzyme, which presents a high degree of oligomerization, was crystallized by vapour-diffusion methods. The crystals belonged to space group P3(1)21, with unit-cell parameters a=268.6, b=268.6, c=224.4?Å. The crystals diffracted to 3.3?Å resolution and gave complete data sets using a synchrotron X-ray source. PMID:23192042

  18. Allantoin transport in Saccharomyces cerevisiae is regulated by two induction systems.

    Science.gov (United States)

    Cooper, T G; Chisholm, V T; Cho, H J; Yoo, H S

    1987-01-01

    We show that the allantoin transport system of Saccharomyces cerevisiae responds to two induction systems, one mediated by allophanate or its analog oxalurate and the other mediated by allantoin or its analog hydantoin acetate. The effects of the two inducers were additive in strain M85. Like other allantoin pathway genes, oxalurate-mediated induction of allantoin transport required a functional DAL81 gene product. Hydantoin acetate-mediated induction of the system, on the other hand, occurred normally in dal81 mutants. This suggests that induction was not only mediated by two separate inducers, but also involved different regulatory proteins. Induction is probably a transcriptionally regulated process, because addition of hydantoin acetate or oxalurate to the culture medium increased the steady-state levels of mRNA encoded by a gene required for allantoin transport (DAL4). Images PMID:2820939

  19. Utilization by Saccharomyces cerevisiae of 5'-methylthioadenosine as a source of both purine and methionine.

    OpenAIRE

    Cone, M. C.; Marchitto, K.; Zehfus, B.; Ferro, A. J.

    1982-01-01

    Cells of the yeast Saccharomyces cerevisiae are normally impermeable to the purine nucleosides adenosine and 5'-deoxy-5'-methylthioadenosine (MTA), a product of polyamine biosynthesis. cordycepin-sensitive, adenosine-utilizing strains of S. cerevisiae were able to use MTA to fulfill an auxotrophic requirement for purine. Cordycepin-sensitive strains carrying a met5 mutation were also able to use MTA as a source of methionine. These MTA-utilizing strains of S. cerevisiae should be useful for m...

  20. Comparison between two selected Saccharomyces cerevisiae strains as fermentation starters in the production of traditional cachaça

    OpenAIRE

    Fátima de Cássia Oliveira Gomes; Roberta Amália de Carvalho Araújo; Patrícia Silva Cisalpino; Elizabeth Spangler Andrade Moreira; Carlos Leomar Zani; Carlos Augusto Rosa

    2009-01-01

    Two Saccharomyces cerevisiae strains were tested as the starter yeasts in a traditional cachaça distillery. The strains used were S. cerevisiae UFMG-A829, isolated from a cachaça fermentation process, and S. cerevisiae K1-V1116, obtained from the wine industry. The permanence of each strain in the fermentation must was determined by RAPD (Random Amplified Polymorphic DNA)-PCR, with primer M13. Both yeast strains were prevalent in the vats for approximately 30 days. Indigenous non-Saccharomy...

  1. Industrial Systems Biology of Saccharomyces cerevisiae Enables Novel Succinic Acid Cell Factory.

    OpenAIRE

    Otero, Jose? Manuel; Cimini, Donatella; Patil, Kiran Raosaheb; Poulsen, Simon Guldberg; Olsson, Lisbeth; Nielsen, Jens

    2013-01-01

    Saccharomyces cerevisiae is the most well characterized eukaryote, the preferred microbial cell factory for the largest industrial biotechnology product (bioethanol), and a robust commerically compatible scaffold to be exploitted for diverse chemical production. Succinic acid is a highly sought after added-value chemical for which there is no native pre-disposition for production and accmulation in S. cerevisiae. The genome-scale metabolic network reconstruction of S. cerevisiae enabled in si...

  2. Development of Efficient Xylose Fermentation in Saccharomyces cerevisiae: Xylose Isomerase as a Key Component:

    OpenAIRE

    Maris, A. J. A.; Winkler, A. A.; Kuyper, M.; Laat, W. T.; Dijken, J. P.; Pronk, J. T.

    2007-01-01

    Metabolic engineering of Saccharomyces cerevisiae for ethanol production from d-xylose, an abundant sugar in plant biomass hydrolysates, has been pursued vigorously for the past 15 years. Whereas wild-type S. cerevisiae cannot ferment d-xylose, the ketoisomer d-xylulose can be metabolised slowly. Conversion of d-xylose into d-xylulose is therefore crucial in metabolic engineering of xylose fermentation by S. cerevisiae. Expression of heterologous xylose reductase and xylitol dehydrogenase doe...

  3. Epidemiological Investigation of Vaginal Saccharomyces cerevisiae Isolates by a Genotypic Method

    OpenAIRE

    Mccullough, Michael J.; Clemons, Karl V.; Farina, Claudio; Mccusker, John H.; Stevens, David A.

    2000-01-01

    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 S. cerevisiae isolates obtained from patients suffering from vaginitis and four isolates...

  4. Ecological Success of a Group of Saccharomyces cerevisiae/Saccharomyces kudriavzevii Hybrids in the Northern European Wine-Making Environment

    OpenAIRE

    Erny, C.; Raoult, P.; Alais, A.; Butterlin, G.; Delobel, P.; Matei-radoi, F.; Casaregola, S.; Legras, J. L.

    2012-01-01

    The hybrid nature of lager-brewing yeast strains has been known for 25 years; however, yeast hybrids have only recently been described in cider and wine fermentations. In this study, we characterized the hybrid genomes and the relatedness of the Eg8 industrial yeast strain and of 24 Saccharomyces cerevisiae/Saccharomyces kudriavzevii hybrid yeast strains used for wine making in France (Alsace), Germany, Hungary, and the United States. An array-based comparative genome hybridization (aCGH) pro...

  5. [Saccharomyces cerevisiae fungemia in an elderly patient following probiotic treatment].

    Science.gov (United States)

    Eren, Zehra; Gurol, Ye?im; Sonmezoglu, Meral; Eren, Hatice Seyma; Celik, Gülden; Kantarci, Gülçin

    2014-04-01

    Saccharomyces cerevisiae, known as baker's yeast, is also used as a probiotic agent to treat gastroenteritis by modulating the endogenous flora and immune system. However, since there have been increasing reports of fungemia due to S.cerevisiae and its subspecies S.boulardii, it is recommended that probiotics should be cautiously used in immunosuppressed patients, people with underlying diseases and low-birth weight babies. To emphasize this phenomenon, in this report, a case of S.cerevisiae fungemia developed in a patient given probiotic treatment for antibiotic-associated diarrhea, was presented. An 88-year-old female patient was admitted to our hospital with left hip pain, hypotension, and confusion. Her medical history included hypertension, chronic renal failure, left knee replacement surgery, and recurrent urinary tract infections due to neurogenic bladder. She was transferred to the intensive care unit with the diagnosis of urosepsis. After obtaining blood and urine samples for culture, empirical meropenem (2 x 500 mg) and linezolid (1 x 600 mg) treatment were administered. A central venous catheter (CVC) was inserted and after one day of inotropic support, her hemodynamic parameters were stabilized. The urine culture obtained on admission yielded extended-spectrum beta-lactamase-producing Klebsiella pneumoniae and Escherichia coli. Urine culture was repeated after three days and no bacteria were isolated. On the 4th day of admission she developed diarrhea. Toxin A/B tests for Clostridium difficile were negative. To relieve diarrhea, S.boulardii (Reflor 250 mg capsules, Sanofi Aventis, Turkey) was administered twice a day, without opening capsules. Two days later, her C-reactive protein (CRP) level increased from 23.2 mg/L to 100 mg/L without fever. Her blood culture taken from the CVC yielded S.cerevisiae. Linezolid and meropenem therapies were stopped on the 13th and 14th days, respectively, while prophylactic fluconazole therapy was replaced with caspofungin 1 x 50 mg on the fifth day. After seven days of therapy CRP and serum creatinine levels decreased to 9.1 mg/L and 1.2 mg/dl, respectively; and she was discharged from the hospital with improvement. The probiotic capsules were used unopen, thus, it was proposed that S.cerevisiae fungemia originated from translocation from the intestinal mucosa. Since it was not possible to investigate the molecular genetics of the strain isolated from the blood culture and the strain present in the probiotic, a definite conclusion about the origin of the strain could not be reached. It was thought that old age and underlying disease of the patient were the related predisposing factors for S.cerevisiae fungemia. This case emphasized that clinicians should be cautious in case of probiotic application even though in encapsulated form, even in immunocompetent patients with a history of long-term hospital stay and use of broad-spectrum antimicrobials since there may be a risk of S.cerevisiae fungemia development. PMID:24819274

  6. The Saccharomyces cerevisiae MADS-box transcription factor Rlm1 is a target for the Mpk1 mitogen-activated protein kinase pathway.

    OpenAIRE

    Dodou, E.; Treisman, R.

    1997-01-01

    Mutation of Saccharomyces cerevisiae RLM1, which encodes a MADS-box transcription factor, confers resistance to the toxic effects of constitutive activity of the Mpk1 mitogen-activated kinase (MAPK) pathway. The Rlm1 DNA-binding domain, which is similar to that of the metazoan MEF2 transcription factors, is also closely related to that of a second S. cerevisiae protein, Smp1 (second MEF2-like protein), encoded by the YBR182C open reading frame (N. Demolis et al., Yeast 10:1511-1525, 1994; H. ...

  7. ACÚMULO DE CÁDMIO POR Saccharomyces cerevisiae FERMENTANDO MOSTO DE MELAÇO

    Scientific Electronic Library Online (English)

    L.G. do, PRADO-FILHO; R.N., DOMINGOS; S.M.G. da, SILVA.

    1998-01-01

    Full Text Available O presente trabalho visou o estudo do acúmulo de cádmio (Cd) por Saccharomyces cerevisiae, fermentando mosto de melaço com contaminações controladas em níveis sub-tóxicos do citado metal. As condições de fermentação foram similares às reinantes na produção industrial de etanol. O mosto, não esterili [...] zado, continha 12% de açúcares redutores totais (ART) e pH 4,5. Para a contaminação controlada empregou-se dois sais de cádmio, cloreto e acetato e, quatro níveis de contaminação 0,5; 1,0; 2,0 e 5,0 mg Cd.kg-1 mosto. A inoculação do mosto foi executada com fermento de panificação (10% p/p). Após a fermentação (4 horas) foram determinados, porcentagem de fermento no vinho centrifugado e teor alcoólico. Na levedura separada foram determinados peso úmido, matéria seca, proteína bruta e teores de cádmio por espectrofotometria de absorção atômica. Em todos os níveis de contaminação estudados houve acúmulo de Cd pela levedura e diminuição do rendimento em etanol. Abstract in english The aim of this paper was to study the cadmium (Cd) accumulation by Saccharomyces cerevisiae fermenting wort of molasses, under sub-toxic levels of controlled cadmium contamination. Fermentation conditions were similar to industrial alcohol production. Non-sterelized wort had 12% of total reducing s [...] ugars (w/w) and pH 4.5. For the controlled contamination, two cadmium salts were used (chloride and acetate), at four levels of contamination: 0.5; 1.0; 2.0 and 5.0 mg Cd.kg-1 wort. The inoculation of the wort was carried out with commercial bread yeast (10% w/w). After fermentation (4 hours), samples were evaluated for cellular viability, alcohol content and yeast percentage in the centrifuged wine. The centrifuged yeast cells were evaluated for total fresh and dry weight, total protein, and cadmium concentration by atomic absortion spectroscopy. In all Cd levels, there was cadmium accumulation by yeast and a decrease in ethanol yield.

  8. ACÚMULO DE CÁDMIO POR Saccharomyces cerevisiae FERMENTANDO MOSTO DE MELAÇO

    Directory of Open Access Journals (Sweden)

    L.G. do PRADO-FILHO

    1998-01-01

    Full Text Available O presente trabalho visou o estudo do acúmulo de cádmio (Cd por Saccharomyces cerevisiae, fermentando mosto de melaço com contaminações controladas em níveis sub-tóxicos do citado metal. As condições de fermentação foram similares às reinantes na produção industrial de etanol. O mosto, não esterilizado, continha 12% de açúcares redutores totais (ART e pH 4,5. Para a contaminação controlada empregou-se dois sais de cádmio, cloreto e acetato e, quatro níveis de contaminação 0,5; 1,0; 2,0 e 5,0 mg Cd.kg-1 mosto. A inoculação do mosto foi executada com fermento de panificação (10% p/p. Após a fermentação (4 horas foram determinados, porcentagem de fermento no vinho centrifugado e teor alcoólico. Na levedura separada foram determinados peso úmido, matéria seca, proteína bruta e teores de cádmio por espectrofotometria de absorção atômica. Em todos os níveis de contaminação estudados houve acúmulo de Cd pela levedura e diminuição do rendimento em etanol.The aim of this paper was to study the cadmium (Cd accumulation by Saccharomyces cerevisiae fermenting wort of molasses, under sub-toxic levels of controlled cadmium contamination. Fermentation conditions were similar to industrial alcohol production. Non-sterelized wort had 12% of total reducing sugars (w/w and pH 4.5. For the controlled contamination, two cadmium salts were used (chloride and acetate, at four levels of contamination: 0.5; 1.0; 2.0 and 5.0 mg Cd.kg-1 wort. The inoculation of the wort was carried out with commercial bread yeast (10% w/w. After fermentation (4 hours, samples were evaluated for cellular viability, alcohol content and yeast percentage in the centrifuged wine. The centrifuged yeast cells were evaluated for total fresh and dry weight, total protein, and cadmium concentration by atomic absortion spectroscopy. In all Cd levels, there was cadmium accumulation by yeast and a decrease in ethanol yield.

  9. Septins localize to microtubules during nutritional limitation in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Vázquez de Aldana Carlos R

    2008-10-01

    Full Text Available Abstract Background In Saccharomyces cerevisiae, nutrient limitation stimulates diploid cells to undergo DNA replication and meiosis, followed by the formation of four haploid spores. Septins are a family of proteins that assemble a ring structure at the mother-daughter neck during vegetative growth, where they control cytokinesis. In sporulating cells, the septin ring disassembles and septins relocalize to the prospore membrane. Results Here, we demonstrate that nutrient limitation triggers a change in the localization of at least two vegetative septins (Cdc10 and Cdc11 from the bud neck to the microtubules. The association of Cdc10 and Cdc11 with microtubules persists into meiosis, and they are found associated with the meiotic spindle until the end of meiosis II. In addition, the meiosis-specific septin Spr28 displays similar behavior, suggesting that this is a common feature of septins. Septin association to microtubules is a consequence of the nutrient limitation signal, since it is also observed when haploid cells are incubated in sporulation medium and when haploid or diploid cells are grown in medium containing non-fermentable carbon sources. Moreover, during meiosis II, when the nascent prospore membrane is formed, septins moved from the microtubules to this membrane. Proper organization of the septins on the membrane requires the sporulation-specific septins Spr3 and Spr28. Conclusion Nutrient limitation in S. cerevisiae triggers the sporulation process, but it also induces the disassembly of the septin bud neck ring and relocalization of the septin subunits to the nucleus. Septins remain associated with microtubules during the meiotic divisions and later, during spore morphogenesis, they are detected associated to the nascent prospore membranes surrounding each nuclear lobe. Septin association to microtubules also occurs during growth in non-fermentable carbon sources.

  10. Point mutations implicate repeated sequences as essential elements of the CYC7 negative upstream site in Saccharomyces cerevisiae.

    OpenAIRE

    Wright, C. F.; Zitomer, R. S.

    1985-01-01

    The transcription of the CYC7 gene of Saccharomyces cerevisiae, encoding the iso-2-cytochrome c protein, is controlled by two upstream regulatory elements, a positive element and a negative element. The nature of the DNA sequences in the negative element were investigated in a two-part approach. The first involved the construction of a CYC7-galK fusion gene which placed the coding sequence of the Escherichia coli galactokinase gene under the regulation of the CYC7 upstream sequences. This fus...

  11. Differential transcriptional activation by v-myb and c-myb in animal cells and Saccharomyces cerevisiae.

    OpenAIRE

    Chen, R. H.; Lipsick, J. S.

    1993-01-01

    The v-myb oncogene and its cellular homolog c-myb encode sequence-specific DNA-binding proteins which regulate transcription from promoters containing Myb-binding sites in animal cells. We have developed a Saccharomyces cerevisiae system to assay transcriptional activation by v-Myb and c-Myb. In yeast strains containing integrated reporter genes, activation was strictly dependent upon both the Myb DNA-binding domain and the Myb recognition element. BAS1, an endogenous Myb-related yeast protei...

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

    OpenAIRE

    Flom, Gary; Weekes, Jared; Johnson, Jill L.

    2005-01-01

    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, CPR7 and YDJ1, and two essential genes, SSL2 and UTP21, not previously linked to Hsp9...

  13. Regulatory elements in the FBP1 promoter respond differently to glucose-dependent signals in Saccharomyces cerevisiae

    OpenAIRE

    Zaragoza, Oscar; Vincent, Olivier; Gancedo, Juana M

    2001-01-01

    In Saccharomyces cerevisiae expression of the fructose-1,6-bisphosphatase-encoding gene, FBP1, is controlled by glucose through the upstream activating sequences UAS1 and UAS2 and the upstream repressing sequence URS1 in its promoter. We have studied the regulation of the proteins that could bind to these elements. We have investigated the role of the putative transcription factors Cat8 and Sip4 in the formation of specific DNA-protein complexes with UAS1 and UAS2, and in the expression of UA...

  14. An essential gene, ESR1, is required for mitotic cell growth, DNA repair and meiotic recombination in Saccharomyces cerevisiae.

    OpenAIRE

    Kato, R.; Ogawa, H.

    1994-01-01

    A new mutant, which was sensitive to both methyl-methanesulfonate (MMS) and ultra-violet light (UV) and defective in meiotic recombination, was isolated from Saccharomyces cerevisiae. The gene, ESR1, was cloned by complementation of the MMS sensitivity of the mutant and found to be essential for cell growth, as the deleted haploid strain was lethal. The ESR1 gene was adjacent to the CKS1 gene on chromosome II and encoded a putative 2368-amino acid protein with a molecular weight of 273 k. The...

  15. GTP hydrolysis controls stringent selection of the AUG start codon during translation initiation in Saccharomyces?cerevisiae

    OpenAIRE

    Huang, Han-kuei; Yoon, Heejeong; Hannig, Ernest M.; Donahue, Thomas F.

    1997-01-01

    We have isolated and characterized two suppressor genes, SUI4 and SUI5, that can initiate translation in the absence of an AUG start codon at the HIS4 locus in Saccharomyces cerevisiae. Both suppressor genes are dominant in diploid cells and lethal in haploid cells. The SUI4 suppressor gene is identical to the GCD11 gene, which encodes the ? subunit of the eIF-2 complex and contains a mutation in the G2 motif, one of the four signature motifs that characterizes this subunit to be a G-protein...

  16. Construction of recombinant industrial Saccharomyces cerevisiae strain with bglS gene insertion into PEP4 locus by homologous recombination*

    OpenAIRE

    Zhang, Qiang; Chen, Qi-he; Fu, Ming-liang; Wang, Jin-ling; Zhang, Hong-bo; He, Guo-qing

    2008-01-01

    The bglS gene encoding endo-l,3-1,4-?-glucanase from Bacillus subtilis was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeast mating pheromone ?-factor (MF?1S), and ADH1 terminator with G418-resistance as the selected marker, was constructed. Then one of the PEP4 allele of Saccharomyces cerevisiae WZ65 strain was replaced by bglS expression cassette using chromosomal integration of polymerase chain ...

  17. Functional domains of a positive regulatory protein, PHO4, for transcriptional control of the phosphatase regulon in Saccharomyces cerevisiae.

    OpenAIRE

    Ogawa, N.; Oshima, Y.

    1990-01-01

    The PHO4 gene encodes a positive regulatory factor involved in regulating transcription of various genes in the phosphatase regulon of Saccharomyces cerevisiae. Besides its own coding region, the 1.8-kilobase PHO4 transcript contains a coding region for a mitochondrial protein which does not appear to be translated. Four functional domains were found in the PHO4 protein, which consists of 312 amino acid (aa) residues as deduced from the open reading frame of PHO4. A gel retardation assay with...

  18. Production of resveratrol from tyrosine in metabolically engineered Saccharomyces cerevisiae.

    Science.gov (United States)

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

    2012-09-10

    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

  19. Variability in individual cell cycles of Saccharomyces cerevisiae.

    Science.gov (United States)

    Lord, P G; Wheals, A E

    1981-08-01

    The kinetics of cell proliferation of Saccharomyces cerevisiae were studied at 4 growth rates using time-lapse cinephotomicrography. Cells were grown on media with a high refractive index to reveal greater intracellular detail under the phase-contrast microscope. The morphological cell-cycle events scored were: bud emergence, nuclear migration, nuclear division, onset of cytokinesis and cell separation. Cell size was measured at cell separation and at bud emergence. The daughter-cycle time was always longer than the parent-cycle time mainly due to the large difference in the lengths of the unbudded phases. Parent cells had a shorter budded period than daughter cells. The large variance in daughter-cycle times was accounted for by the large variance in the lengths of the unbudded phase of daughter cells. The duration and variability of the periods in the cyclc from nuclear migration onwards were equivalent for parent and daughter cells. Daughter cells were always smaller than parent cells at division. There was wide variation in cell size at both division and bud emergence. The results indicated that a modified deterministic model could best explain cell proliferation kinetics in yeast. The data were used to evaluate 2 different models. The 'sloppy size control' model of Wheals (1981 a) was more consistent with the data than the 'tandem' model of Shilo, Shilo & Simchen (1976). The distribution of unbudded periods of daughter cells suggested that there was an additional incompressible period not present in parent cells. PMID:7033253

  20. Size control models of Saccharomyces cerevisiae cell proliferation.

    Science.gov (United States)

    Wheals, A E

    1982-04-01

    By using time-lapse photomicroscopy, the individual cycle times and sizes at bud emergence were measured for a population of saccharomyces cerevisiae cells growing exponentially under balanced growth conditions in a specially constructed filming slide. There was extensive variability in both parameters for daughter and parent cells. The data on 162 pairs of siblings were analyzed for agreement with the predictions of the transition probability hypothesis and the critical-size hypothesis of yeast cell proliferation and also with a model incorporating both of these hypotheses in tandem. None of the models accounted for all of the experimental data, but two models did give good agreement to all of the data. The wobbly tandem model proposes that cells need to attain a critical size, which is very variable, enabling them to enter a start state from which they exit with first order kinetics. The sloppy size control model suggests that cells have an increasing probability per unit time of traversing start as they increase in size, reaching a high plateau value which is less than one. Both models predict that the kinetics of entry into the cell division sequence will strongly depend on variability in birth size and thus will be quite different for daughters and parents of the asymmetrically dividing yeast cells. Mechanisms underlying these models are discussed. PMID:7050671

  1. BIOTECHNOLOGICAL PRODUCTION OF ETHANOL BY SACCHAROMYCES CEREVISIAE, USING DIFFERENT SUBSTRATES

    Directory of Open Access Journals (Sweden)

    Vijaya S. Gulalkayi

    2012-12-01

    Full Text Available Research efforts are needed to design and improve the process, which would produce sustainable and economically feasible transportation fuel. The present investigation was undertaken to determine the availability of carbohydrates in hydrolysates derived from different substrates Acacia arabica, Delbergia sisso, Peltophorum Pterocarpur and Perkia biglobosa pods, in the production of ethanol. The enzymatic hydrolysis of the substrates has yielded the significant amount of reducing sugar from the substrates by comparing the effect of enzymes on hydrolysis. The Acacia arabica pods has showed the higher production of reducing sugars when treated with 4% a-amylase whereas Peltophorum pterocarpum has produced lowest yield of reducing sugar at 4% a-amylase enzyme. The optimum temperature required for the activity of a-amylase enzyme in the production of reducing sugars using different substrates were revealed that at 30oC the Acacia arabica has yielded maximum sugars whereas the Perkia biglobosa has showed the minimum yield of reducing sugar. The optimum period of enzyme activity in the production of reducing sugars using different substrates was indicted that the Acacia arabica has showed the maximum yield of reducing sugars during the incubation period of 24 hours whereas minimum yield was observed in Perkia biglobosa. The optimum incubation period of Saccharomyces cerevisiae in the production of ethanol has showed that the seven days of incubation has yielded maximum amount of ethanol using the substrate Acacia arabica.

  2. Biosynthesis of diphthamide in the yeast Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Inactivation of EF-2 by diphtheria toxin requires the presence of a posttranslationally synthesized amino acid residue, diphthamide. The present work was undertaken to study the biosynthetic mechanism of diphthamide synthesis in the yeast Saccharomyces cerevisiae in order to gain better understanding of the biological roles of this unique amino acid residue. Thirty-one haploid ADP-ribosylation-negative mutants, comprising 5 complementation groups, were obtained. One of these mutants contains a toxin-resistant form of EF-2 which can be converted to a toxin-sensitive form through the methylation reaction catalyzed by a S-AdoMet:EF-2 methyltransferase enzyme which is present in other yeast strains. The [3He]methylated residue in the EF-2 modified by the methyltransferase in the presence of S-Ado-L-[3H-methyl]-Met has been analyzed chromatographically following both acid and enzymatic hydrolysis. At the conclusion of the reaction, all of the radiolabel was recovered as diphthine (the unamidated form of diphthamide). The authors conclude that the S-AdoMet:EF-2-methyltransferase is specific for the addition of at least the last two of the three methyl groups present in diphthine

  3. Passage through stationary phase advances replicative aging in Saccharomyces cerevisiae

    Science.gov (United States)

    Ashrafi, Kaveh; Sinclair, David; Gordon, Jeffrey I.; Guarente, Leonard

    1999-01-01

    Saccharomyces cerevisiae mother cells undergo an aging program that includes morphologic changes, sterility, redistribution of the Sir transcriptional silencing complex from HM loci and telomeres to the nucleolus, alterations in nucleolar architecture, and accumulation of extrachromosomal ribosomal DNA circles (ERCs). We report here that cells starved for nutrients during prolonged periods in stationary phase show a decrease in generational lifespan when they reenter the cell cycle. This shortened lifespan is not transmitted to progeny cells, indicating that it is not due to irreversible genetic damage. The decrease in the lifespan is accompanied by all of the changes of accelerated aging with the notable exception that ERC accumulation is not augmented compared with generation-matched, nonstarved cells. These results suggest a number of models, including one in which starvation reveals a component of aging that works in parallel with the accumulation of ERCs. Stationary-phase yeast cells may be a useful system for identifying factors that affect aging in other nondividing eukaryotic cells. PMID:10430902

  4. Generation and maintenance of synchrony in Saccharomyces cerevisiae continuous culture.

    Science.gov (United States)

    Murray, Douglas B; Klevecz, Robert R; Lloyd, David

    2003-07-01

    Cultures of Saccharomyces cerevisiae grown continuously produce an autonomous oscillation in many metabolic outputs. The most conveniently measured variable, i.e., dissolved oxygen concentration, oscillates with a period of 40-55 min. Previously we have identified two compounds capable of resetting phase, acetaldehyde and hydrogen sulfide. The phase-response curves constructed for acetaldehyde show a strong (Type 0) response at 3.0 mM and a weak (Type 1) response at 1.0 mM. Ammonium sulfide phase-response curves (pulse injected at 1.0 microM and 3.0 microM) revealed that sulfide is only an effective perturbation agent when endogenous sulfide concentrations are at a maximum. Also only Type 1 phase responses were observed. When the phase-response curve for sulfite (at 3.0 M) was constructed, phase responses were at a maximum at 60 degrees, indicating the possible involvement of sulfite in cell synchronization. It is concluded that endogenously produced acetaldehyde and sulfite tune the oscillation of mitochondrial energization state whereas sulfide mediates population synchrony. PMID:12799177

  5. Nitrogen Repression of the Allantoin Degradative Enzymes in Saccharomyces cerevisiae

    Science.gov (United States)

    Bossinger, June; Lawther, Robert P.; Cooper, Terrance G.

    1974-01-01

    Saccharomyces cerevisiae can utilize allantoin as a sole nitrogen source by degrading it to ammonia, “CO2,” and glyoxylate. We have previously shown that synthesis of the allantoin degradative enzymes is contingent upon the presence of allophanate, the last intermediate in the pathway. The reported repression of arginase by ammonia prompted us to ascertain whether or not the allantoin degradative system would respond in a similar manner. We observed that the differential rates of allantoinase and allophanate hydrolase synthesis were not decreased appreciably when comparing cultures grown on urea to those grown on urea plus ammonia. These experiments were also performed using the strain and conditions previously reported by Dubois, Grenson, and Wiame. We found allophanate hydrolase production to be twofold repressed by ammonia when that strain was grown on glucose-urea plus ammonia medium. If, however, serine or a number of other readily metabolized amino acids were provided in place of ammonia, production of the allantoin degradative enzymes was quickly (within 20 min) and severely repressed in both strains. We conclude that repression previously attributed to ammonia may result from its metabolism to amino acids and other metabolites. PMID:4598006

  6. Genotoxicity assessment of amaranth and allura red using Saccharomyces cerevisiae.

    Science.gov (United States)

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

    2013-01-01

    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

  7. Heterologous Expression of Syntaxin 6 in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    MARTIN GÖTTE

    2002-01-01

    Full Text Available The molecular mechanisms of vesicular protein transport in eukaryotic cells are highly conserved. Members of the syntaxin family play a pivotal role in the membrane fusion process. We have expressed rat syntaxin 6 and its cytoplasmic domain in wild-type and pep12 mutant strains of Saccharomyces cerevisiae to elucidate the role of the syntaxin 6-dependent vesicular trafficking step in yeast. Immunofluorescence microscopy revealed a punctate, Golgi-like staining pattern for syntaxin 6, which only partially overlapped with Pep12p in wild-type yeast cells. In contrast to Pep12p, syntaxin 6 was not mislocalized to the vacuole upon expression from 2 micron vectors, which might be attributed to conserved sorting and retention signals. Syntaxin 6 was not capable of complementing the sorting and maturation defects of the vacuolar hydrolase CPY in pep12 null mutants. No dominant negative effects of either syntaxin 6 or syntaxin 6deltaC overexpression on CPY sorting and maturation were observed in wild-type yeast cells. We conclude that syntaxin 6 and Pep12p do not act at the same vesicular trafficking step(s in yeast and higher eukaryotes

  8. Antimutators of mitochodrial and nuclear DNA in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    In Saccharomyces cerevisiae ten antimutator mutants have been isolated. The spontaneous occurrence of mitochondrial mutants resistant to erythromycin, oligomycin, and diuron is decreased 2-60-fold in these strains. The rate of forward and reverse spontaneous mutations of the nuclear genome is also reduced. The meiotic progenies arising from the crosses of seven mutants (LB1, LB2, LB4, LB5, LB6, LB7, LB10) with an isogenic parental strain exhibit 2:2 segregations and therefore are the result of mutations in a single nuclear gene. The six mutants LB1, LB2, LB4, LB6, LB7, LB10 are semidominant and determine six complementation groups. The mutant LB5 is dominant and therefore cannot be assigned to any complementation group. The mutants. LB1, LB4 and LB10 are gamma-ray sensitive and, by tetrad analysis, it has been shown that gamma-ray sensitivity and spontaneous antimutability are the result of a single nuclear gene mutation. The other three mutants LB3, LB8 and LB9 exhibit complex tetrad segregations, typical of cytoplasmic inheritance and do not complement each other. However, although the mutations are semidominant, it has not been possible to detect any antimutator cytoductant among some 500 cytoductants carrying the karl 1-1 nucleus. (orig./AJ)ng the karl 1-1 nucleus. (orig./AJ)

  9. Metabolic impact of redox cofactor perturbations in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Hou, Jin; Lages, Nuno

    2009-01-01

    Redox cofactors play a pivotal role in coupling catabolism with anabolism and energy generation during metabolism. There exists a delicate balance in the intracellular level of these cofactors to ascertain an optimal metabolic output. Therefore, cofactors are emerging to be attractive targets to induce widespread changes in metabolism. We present a detailed analysis of the impact of perturbations in redox cofactors in the cytosol or mitochondria on glucose and energy metabolism in Saccharomyces cerevisiae to aid metabolic engineering decisions that involve cofactor engineering. We enhanced NADH oxidation by introducing NADH oxidase or alternative oxidase, its ATP-mediated conversion to NADPH using NADH kinase as well as the interconversion of NADH and NADPH independent of ATP by the soluble, non-proton-translocating bacterial transhydrogenase. Decreasing cytosolic NADH level lowered glycerol production, while decreasing mitochondrial NADH lowered ethanol production. However, when these reactions were coupled with NADPH production, the metabolic changes were more moderated. The direct consequence of these perturbations could be seen in the shift of the intracellular concentrations of the cofactors. The changes in product profile and intracellular metabolite levels were closely linked to the ATP requirement for biomass synthesis and the efficiency of oxidative phosphorylation, as estimated from a simple stoichiometric model. The results presented here will provide valuable insights for a quantitative understanding and prediction of cellular response to redox-based perturbations for metabolic engineering applications.

  10. Effects of low X-ray doses in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Three strains of Saccharomyces cerevisiae with different capacities for repair of radiation damage (RAD, rad18, and rad52) have been tested for their colony forming ability (CFA) and growth rates after application of small X-ray doses from 3.8 mGy to 40 Gy. There was no reproducible increase in CFA observable after application of doses between 3.8 mGy and 4.7 Gy.X-ray doses of 40 Gy causing an inactivation of CFA from 90% to 50%, depending on the repair capacity of the strains used, caused a reduced increase in optical density during 2 h buffer treatment in comparison to unirradiated cells. This reduction however, is reversible as soon as the cells are transferred into nutrient medium. One hour after transfer into growh medium the portions of cells with large buds (Gs and M phase) and cells with small buds (S phase) are drastically different in irradiated cells from those obtained in unirradiated cells. The time necessary for separation of mother and daughter cells is prolonged by X-ray irradiation and the formation of new buds is retarded. (orig.)

  11. MAP kinase pathways in the yeast Saccharomyces cerevisiae

    Science.gov (United States)

    Gustin, M. C.; Albertyn, J.; Alexander, M.; Davenport, K.; McIntire, L. V. (Principal Investigator)

    1998-01-01

    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.

  12. Fermentação de trealose e glicogênio endógenos em Saccharomyces cerevisiae Fermentation of endogenous trehalose and glycogen by Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    L.V. FERREIRA

    1999-01-01

    Full Text Available As linhagens PE-2 e VR-1 de Saccharomyces cerevisiae foram submetidas à fermentação das reservas endógenas na temperatura de 40oC. No intervalo de 0 a 24 horas foram recolhidas as amostras para a determinação de etanol, nitrogênio no fermento e no vinho, bem como os carboidratos de reserva (trealose e glicogênio e a viabilidade celular. A trealose foi esgotada durante 24 horas. Os teores de glicogênio sofreram muitas oscilações ao longo do tempo, entre a mobilização e a síntese e embora não esgotado, deve ter contribuído significativamente para a formação de álcool na suspensão. Foi observada a relação proporcional entre a mobilização de trealose e a queda da viabilidade celular. No transcorrer da fermentação das reservas de carboidratos houve aumento nos teores de nitrogênio no fermento até 6 e 8 horas, sendo tal incremento afetado pela linhagem de levedura. No prosseguimento da fermentação ocorreu a autólise celular, que foi percebida pelo aumento brusco de nitrogênio no vinho (de 200 para 1500mg/L e pela queda da viabilidade celular. O ganho alcançado com a fermentação endógena foi de 40 e 68 litros de álcool por tonelada de levedura seca com incremento de 25 e 27% de proteína no fermento para as linhagens PE-2 e VR-1, respectivamente. Este resultado tem reflexos positivos quando da comercialização da levedura seca como proteína microbiana.Two Saccharomyces cerevisiae strains (PE-2 and VR-1 were subjected to fermentation of its carbohidrate reserve (Trehalose and glycogen at 40oC. During a 24 hours interval samples were collected for determination of ethanol, yeast and wine nitrogen, yeast trehalose, glycogen and cell viability. Trehalose was completely exhausted after 24 hours. Glycogen was not completely consumed, but probably contributes for ethanol formation. As trehalose is consumed yeast cell viability decreases, while yeast nitrogen content increase, reaching a maximum between 6 and 8 hours, depending on the yeast strain. If yeast is maintained under prolonged stressing conditions, cell autolysis occurs and nitrogen is lost to the medium, increasing from 200 to 1500mg/L. Such endogenous fermentation allows a production of 40 to 68L of ethanol per ton of dry yeast, with yeast nitrogen increasing of 25 and 27% for PE-2 and VR-1, respectively.

  13. Fermentação de trealose e glicogênio endógenos em Saccharomyces cerevisiae / Fermentation of endogenous trehalose and glycogen by Saccharomyces cerevisiae

    Scientific Electronic Library Online (English)

    L.V., FERREIRA; H.V., AMORIM; L.C., BASSO.

    1999-01-01

    Full Text Available As linhagens PE-2 e VR-1 de Saccharomyces cerevisiae foram submetidas à fermentação das reservas endógenas na temperatura de 40oC. No intervalo de 0 a 24 horas foram recolhidas as amostras para a determinação de etanol, nitrogênio no fermento e no vinho, bem como os carboidratos de reserva (trealose [...] e glicogênio) e a viabilidade celular. A trealose foi esgotada durante 24 horas. Os teores de glicogênio sofreram muitas oscilações ao longo do tempo, entre a mobilização e a síntese e embora não esgotado, deve ter contribuído significativamente para a formação de álcool na suspensão. Foi observada a relação proporcional entre a mobilização de trealose e a queda da viabilidade celular. No transcorrer da fermentação das reservas de carboidratos houve aumento nos teores de nitrogênio no fermento até 6 e 8 horas, sendo tal incremento afetado pela linhagem de levedura. No prosseguimento da fermentação ocorreu a autólise celular, que foi percebida pelo aumento brusco de nitrogênio no vinho (de 200 para 1500mg/L) e pela queda da viabilidade celular. O ganho alcançado com a fermentação endógena foi de 40 e 68 litros de álcool por tonelada de levedura seca com incremento de 25 e 27% de proteína no fermento para as linhagens PE-2 e VR-1, respectivamente. Este resultado tem reflexos positivos quando da comercialização da levedura seca como proteína microbiana. Abstract in english Two Saccharomyces cerevisiae strains (PE-2 and VR-1) were subjected to fermentation of its carbohidrate reserve (Trehalose and glycogen) at 40oC. During a 24 hours interval samples were collected for determination of ethanol, yeast and wine nitrogen, yeast trehalose, glycogen and cell viability. Tre [...] halose was completely exhausted after 24 hours. Glycogen was not completely consumed, but probably contributes for ethanol formation. As trehalose is consumed yeast cell viability decreases, while yeast nitrogen content increase, reaching a maximum between 6 and 8 hours, depending on the yeast strain. If yeast is maintained under prolonged stressing conditions, cell autolysis occurs and nitrogen is lost to the medium, increasing from 200 to 1500mg/L. Such endogenous fermentation allows a production of 40 to 68L of ethanol per ton of dry yeast, with yeast nitrogen increasing of 25 and 27% for PE-2 and VR-1, respectively.

  14. Localization of nuclear retained mRNAs in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Thomsen, Rune; Libri, Domenico

    2003-01-01

    In the yeast Saccharomyces cerevisiae, a common conditional phenotype associated with deletion or mutation of genes encoding mRNA export factors is the rapid accumulation of mRNAs in intranuclear foci, suggested to be near transcription sites. The nuclear RNA exosome has been implicated in retaining RNAs in these foci; on deletion of the exosome component Rrp6p, the RNA is released. To determine the exact nuclear location of retained as well as released mRNAs, we have used mRNA export mutant strains to analyze the spatial relationship between newly synthesized heat shock mRNA, the chromosomal site of transcription, and known S. cerevisiae nuclear structures such as the nucleolus and the nucleolar body. Our results show that retained SSA4 RNA localizes to an area in close proximity to the SSA4 locus. On deletion of Rrp6p and release from the genomic locus, heat shock mRNAs produced in the rat7–1 strain colocalize predominantly with nucleolar antigens. Bulk poly(A)+ RNA, on the other hand, is localized primarily to the nuclear rim. Interestingly, the RNA binding nucleocytoplasmic shuttle protein Npl3p shows strong colocalization with bulk poly(A)+ RNA, regardless of its nuclear location. Taken together, our data show that retention occurs close to the gene and indicate distinct nuclear fates of different mRNAs.

  15. The role of cell wall revealed by the visualization of Saccharomyces cerevisiae transformation.

    Science.gov (United States)

    Pham, Tuan Anh; Kawai, Shigeyuki; Kono, Emi; Murata, Kousaku

    2011-03-01

    Transformation is an indispensable method for the manipulation of Saccharomyces cerevisiae cell. The spf1 cell, in which the gene encoding an endoplasmic reticulum-located P-type ATPase is deleted, has been known to show the high-transformation phenotype. In this study, fluorescent microscopic observation of transformation process of S. cerevisiae using plasmid DNA labelled with fluorescent DNA probe, YOYO-1, suggested that the spf1 cell absorbed more plasmid DNA on cellular surface than did the wild-type cell and the unwashed cell did more plasmid DNA than the washed cell. The amounts of the absorbed DNA correlated with the transformation efficiency (number of transformants per ?g plasmid DNA) and frequency (transformation efficiency per viable cell number). The high-transformation phenotype of spf1 cell and the effect of heat shock, which effectively induces the transformation of intact cell, disappeared upon cell wall digestion. Electron microscopic observation of the transformation process using negatively charged Nanogold as a mimic of plasmid DNA supported the result obtained using YOYO-1 and implied that plasmid DNA enters into cell together with membrane structure. These data strongly suggest that during the transformation of intact cell, plasmid DNA is initially absorbed on the cell wall, passes through the cell wall with the aid of heat shock, reaches to the membrane, and enters into the cell together with the membrane structure and that the capacity of the cell wall to absorb DNA is at least one of the determinants of transformation efficiency and frequency. PMID:21079962

  16. Overexpression of NADH-dependent fumarate reductase improves D-xylose fermentation in recombinant Saccharomyces cerevisiae.

    Science.gov (United States)

    Salusjärvi, Laura; Kaunisto, Sanna; Holmström, Sami; Vehkomäki, Maija-Leena; Koivuranta, Kari; Pitkänen, Juha-Pekka; Ruohonen, Laura

    2013-12-01

    Deviation from optimal levels and ratios of redox cofactors NAD(H) and NADP(H) is common when microbes are metabolically engineered. The resulting redox imbalance often reduces the rate of substrate utilization as well as biomass and product formation. An example is the metabolism of D-xylose by recombinant Saccharomyces cerevisiae strains expressing xylose reductase and xylitol dehydrogenase encoding genes from Scheffersomyces stipitis. This pathway requires both NADPH and NAD(+). The effect of overexpressing the glycosomal NADH-dependent fumarate reductase (FRD) of Trypanosoma brucei in D-xylose-utilizing S. cerevisiae alone and together with an endogenous, cytosol directed NADH-kinase (POS5?17) was studied as one possible solution to overcome this imbalance. Expression of FRD and FRD + POS5?17 resulted in 60 and 23 % increase in ethanol yield, respectively, on D-xylose under anaerobic conditions. At the same time, xylitol yield decreased in the FRD strain suggesting an improvement in redox balance. We show that fumarate reductase of T. brucei can provide an important source of NAD(+) in yeast under anaerobic conditions, and can be useful for metabolic engineering strategies where the redox cofactors need to be balanced. The effects of FRD and NADH-kinase on aerobic and anaerobic D-xylose and D-glucose metabolism are discussed. PMID:24113892

  17. Trs85 is Required for Macroautophagy, Pexophagy and Cytoplasm to Vacuole Targeting in Yarrowia lipolytica and Saccharomyces cerevisiae

    Science.gov (United States)

    Nazarko, Taras Y.; Huang, Ju; Nicaud, Jean-Marc; Klionsky, Daniel J.; Sibirny, Andrei A.

    2006-01-01

    Yarrowia lipolytica was recently introduced as a new model organism to study peroxisome degradation in yeasts. Transfer of Y. lipolytica cells from oleate/ethylamine to glucose/ammonium chloride medium leads to selective macroautophagy of peroxisomes. To decipher the molecular mechanisms of macropexophagy we isolated mutants of Y. lipolytica defective in the inactivation of peroxisomal enzymes under pexophagy conditions. Through this analysis we identified the gene YlTRS85, the ortholog of Saccharomyces cerevisiae TRS85 that encodes the 85 kDa subunit of transport protein particle (TRAPP). A parallel genetic screen in S. cerevisiae also identified the trs85 mutant. Here, we report that Trs85 is required for nonspecific autophagy, pexophagy and the cytoplasm to vacuole targeting pathway in both yeasts. PMID:16874038

  18. Effects of Saccharomyces cerevisiae Yeast on Tibia Bone Characteristics in Rabbits

    Directory of Open Access Journals (Sweden)

    Ilker Arican

    2012-01-01

    Full Text Available The effect of feeding different levels of yeast Saccharomyces cerevisiae on tibial bone characteristics of the New Zealand rabbits was studied. An experiment of 85 days duration was conducted with male 5-6 weeks old rabbits. There were 3 dietary treatments each consisting of 7 rabbits in each. The treatments were containing 0 (control, 2 and 4 g kg-1 yeast (Saccharomyces cerevisiae in their diet, respectively. At the end of the experiment the right tibia of rabbits were dissected from the surrounding tissues and used for bone force and stress measurements. Results show that there is no effect of feeding with different level of Saccharomyces cerevisiae yeast on body weight, tibia weight and length, bone force and stress values of tibia of the rabbits.

  19. Cellular responses of Saccharomyces cerevisiae to DNA damage

    International Nuclear Information System (INIS)

    Full text. Several experimental strategies have been used to study responses of S. cerevisiae cells to DNA damage. One approach was based on the isolation of novel genes, the expression of which is induced by lesions in DNA. One of these genes, DIN7, was cloned and partially characterized previously. The product of DIN7 belongs to a large family of proteins involved in DNA repair and mutagenesis. This family includes Rad2, Rad27 and ExoI proteins of S. cerevisiae and their respective human homologues, all of which are endowed with DNA nuclease activity. To study cellular function of Din7 we constructed the pPK3 plasmid carrying DIN7 fused to the GAL1 promoter. Effects of DIN7 overproduction on the phenotypes of wild-type cells and of rad27 and exoI mutants were examined. Overproduction of Din7 does not seem to affect the proficiency of wild-type S. cerevisiae cells in recombination and mutagenesis. Also, overexpression of DIN7 does not suppress the deficiency of the EXOI gene product, the closest homologue of Din7, both in recombination and in controlling the fidelity of DNA replication. Unexpectedly, we found that elevated levels of Din7 result in a very high frequency of mitochondrial rho- mutants. A high frequency of production of rho- mutants wa s also observed in strains defective in the functioning of the Dun1 protein kinase involved in signal transmission in cells exposed to DNA damaging agents. Interestingly, deficiency of Dun1 results also in a significant derepression of the DIN7 gene. Experiments are under way to distinguish whether a high cellular level of Din7 specifically decreases stability of mitochondrial DNA or affects stability of chromosomal DNA as well. Analysis of previously constructed S. cerevisiae strains carrying random geno mic fusions with reporter lacZ gene, allowed us to identify the reading frame YBR173c, on chromosome II as a novel damage inducible gene - DIN8. We have shown that DIN8-lacZ fusion is induced in yeast cells treated with MMS or exposed to UV light. Northern RNA analysis indicates that DIN8 is induced in response to DNA damage at the transcriptional level. DIN8 was cloned and the phenotype of cells with disruption of the gene is under study. POL2-MEC1-RAD53-DUN1-signal transducing pathway has recently been postulated to be involved in the regulation of response of S. cerevisiae cells to DNA-damaging agents. We analyzed the expression of a known damage inducible DNA-repair gene, MAG1, encoding 3-methyladenine glycosylase, in S. cerevisiae strains carrying MAG1 ::lacZ fusion and deficient in either POL2, MEC1, RAD53 or DUN1 function. ?-galactosidase activity was assayed in cycling cells exposed to MMS or UV light. It was found that, in contrast to model DNA damage inducible RNR genes, neither mutation in t he sensory C-terminal part of polymerase ? (pol2-11) nor the in the Mec1, Sad1/Rad53 or Dun1 cellular kinases blocks the induction of MAG1 in response to MMS or UV light in cycling yeasts. (author)

  20. Analysis of HIV-1 Vpr determinants responsible for cell growth arrest in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Yao Xiao-Jian

    2004-08-01

    Full Text Available Abstract Background The HIV-1 genome encodes a well-conserved accessory gene product, Vpr, that serves multiple functions in the retroviral life cycle, including the enhancement of viral replication in nondividing macrophages, the induction of G2 cell-cycle arrest, and the modulation of HIV-1-induced apoptosis. We previously reported the genetic selection of a panel of di-tryptophan (W-containing peptides capable of interacting with HIV-1 Vpr and inhibiting its cytostatic activity in Saccharomyces cerevisiae (Yao, X.-J., J. Lemay, N. Rougeau, M. Clément, S. Kurtz, P. Belhumeur, and E. A. Cohen, J. Biol. Chem. v. 277, p. 48816–48826, 2002. In this study, we performed a mutagenic analysis of Vpr to identify sequence and/or structural determinants implicated in the interaction with di-W-containing peptides and assessed the effect of mutations on Vpr-induced cytostatic activity in S. cerevisiae. Results Our data clearly shows that integrity of N-terminal ?-helix I (17–33 and ?-helix III (53–83 is crucial for Vpr interaction with di-W-containing peptides as well as for the protein-induced cytostatic effect in budding yeast. Interestingly, several Vpr mutants, mainly in the N- and C-terminal domains, which were previously reported to be defective for cell-cycle arrest or apoptosis in human cells, still displayed a cytostatic activity in S. cerevisiae and remained sensitive to the inhibitory effect of di-W-containing peptides. Conclusions Vpr-induced growth arrest in budding yeast can be effectively inhibited by GST-fused di-W peptide through a specific interaction of di-W peptide with Vpr functional domain, which includes ?-helix I (17–33 and ?-helix III (53–83. Furthermore, the mechanism(s underlying Vpr-induced cytostatic effect in budding yeast are likely to be distinct from those implicated in cell-cycle alteration and apoptosis in human cells.

  1. Removal of lead, mercury and nickel using the yeast Saccharomyces cerevisiae / Remoción de plomo, mercurio y níquel utilizando la levadura Saccharomyces cerevisiae

    Scientific Electronic Library Online (English)

    Cherlys, Infante J; Deniles, De Arco R; Edgardo, Angulo M.

    2014-05-01

    Full Text Available Objetivos. En este estudio se utilizó la biomasa de la levadura Saccharomyces cerevisiae para retener plomo, mercurio y níquel en forma de iones disueltos en agua. Materiales y métodos. Se prepararon soluciones sintéticas que contenían los tres metales pesados, las cuales se pusieron en contacto con [...] el microorganismo en forma viable a diferentes condiciones de pH, temperatura, aireación y agitación. Resultados. Tanto las variables individuales como los efectos de interacción influyeron sobre el proceso de biosorción. A través de todos los experimentos, se observó que la biomasa de Saccharomyces cerevisiae eliminó un mayor porcentaje de plomo (86.4%) en comparación al mercurio y al níquel (69.7 y 47.8% respectivamente). Cuando el pH se fijó en valor de 5, el efecto fue positivo para los tres metales. Conclusiones. El pH fue la variable que tuvo una mayor influencia en la biosorción de plomo sobre la biomasa de Saccharomyces cerevisiae. La afinidad de los metales pesados por la biomasa siguió el orden Pb>Hg>Ni. Abstract in english Objective. In this study the biomass of the yeast Saccharomyces cerevisiae was used to remove lead, mercury and nickel in the form of ions dissolved in water. Materials and methods. Synthetic solutions were prepared containing the three heavy metals, which were put in contact with viable microorgani [...] sms at different conditions of pH, temperature, aeration and agitation. Results. Both individual variables and the interaction effects influenced the biosorption process. Throughout the experimental framework it was observed that the biomass of Saccharomyces cerevisiae removed a higher percentage of lead (86.4%) as compared to mercury and nickel (69.7 and 47.8% respectively). When the pH was set at a value of 5 the effect was positive for all three metals. Conclusions. pH was the variable that had a greater influence on the biosorption of lead on the biomass of Saccharomyces cerevisiae. The affinity of the heavy metals for the biomass followed the order Pb>Hg>Ni.

  2. Prevalence reduction of pathogens in poultry fed with Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Fanelli, A.

    2015-01-01

    Full Text Available Description of the subject. The growth of new antibiotic-resistant strains of pathogens represents a huge problem in poultry rearing. There is evidence that dietary yeast could be effective in the protection against a variety of pathogens that can affect poultry health and cause foodborne diseases in humans. Since still few or contradictory information are available for this topic. Objectives. The objective of this study was to investigate the effects of live yeast supplementation in broiler chickens on Salmonella enteritidis and Campylobacter jejuni content in feces, cecum, and skin. Method. Supplemented yeast consisted of Saccharomyces cerevisiae (Levucell® SB20, type boulardii I-1079, Lallemand, France and was administered at a rate of 1 x 106 CFU·g-1 of feed. On day ten of life, birds were orally challenged with S. enteritidis (1 x 105 CFU/bird and C. jejuni (3 x 105 CFU/bird. Growth performance, and coliforms, yeasts and lactobacilli enumeration were evaluated on day 0, 10, 20 and 38. Ten and eighteen days post infection (PI, 10 animals per replicate were slaughtered and pooled ceca content were analyzed for yeast enumeration and Salmonella and Campylobacter frequency and enumeration. The presence and the enumeration of Salmonella and Campylobacter in neck and breast skin were performed on one subject per replicate. Results. Dietary S. cerevisiae increased yeast and lactobacilli (p = 0.01 count, while Salmonella enumeration and frequency significantly decreased in neck (p = 0.03 and tended to decrease in cecum (p = 0.06, feces (p = 0.06, and breast (p = 0.08. On 10d PI Campylobacter presence was decreased in cecum (p = 0.01, feces (p < 0.01, breast skin (p = 0.04 and neck skin (p < 0.01, while the enumeration was found to be lower in feces (p < 0.01 and neck skin (p = 0.05. At the end of the trial the frequency of this pathogen was decreased in feces (p < 0.01, and breast skin (p = 0.02, while the enumeration was diminished in cecum (p < 0.05 and feces (p < 0.05. Conclusions. The present study shows that the inclusion of Levucell® SB20 can significantly control Campylobacter carriage in chickens with some positive effects also on Salmonella presence, thus reducing the contamination of carcasses at slaughtering and preventing human foodborne diseases.

  3. Effects of Saccharomyces cerevisiae Yeast on Tibia Bone Characteristics in Rabbits

    OpenAIRE

    Ilker Arican

    2012-01-01

    The effect of feeding different levels of yeast Saccharomyces cerevisiae on tibial bone characteristics of the New Zealand rabbits was studied. An experiment of 85 days duration was conducted with male 5-6 weeks old rabbits. There were 3 dietary treatments each consisting of 7 rabbits in each. The treatments were containing 0 (control), 2 and 4 g kg-1 yeast (Saccharomyces cerevisiae) in their diet, respectively. At the end of the experiment the right tibia of rabbits were dissected from the s...

  4. Links between replication and recombination in Saccharomyces cerevisiae: A hypersensitive requirement for homologous recombination in the absence of Rad27 activity

    OpenAIRE

    Debrauwère, Hélène; Loeillet, Sophie; Lin, Waka; Lopes, Judith; Nicolas, Alain

    2001-01-01

    The RAD27 gene of Saccharomyces cerevisiae encodes a 5?-3? flap exo/endonuclease, which plays an important role during DNA replication for Okazaki fragment maturation. Genetic studies have shown that RAD27 is not essential for growth, although rad27? mutants are temperature sensitive. Moreover, they exhibit increased sensitivity to alkylating agents, enhanced spontaneous recombination, and repetitive DNA instability. The conditional lethality conferred by the rad27? mutation indicates t...

  5. Xylulokinase Overexpression in Two Strains of Saccharomyces cerevisiae Also Expressing Xylose Reductase and Xylitol Dehydrogenase and Its Effect on Fermentation of Xylose and Lignocellulosic Hydrolysate

    OpenAIRE

    Johansson, Bjo?rn; Christensson, Camilla; Hobley, Timothy; Hahn-ha?gerdal, Ba?rbel

    2001-01-01

    Fermentation of the pentose sugar xylose to ethanol in lignocellulosic biomass would make bioethanol production economically more competitive. Saccharomyces cerevisiae, an efficient ethanol producer, can utilize xylose only when expressing the heterologous genes XYL1 (xylose reductase) and XYL2 (xylitol dehydrogenase). Xylose reductase and xylitol dehydrogenase convert xylose to its isomer xylulose. The gene XKS1 encodes the xylulose-phosphorylating enzyme xylulokinase. In this study, we dete...

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

    OpenAIRE

    Feldman, D; Tökés, L G; Stathis, P A; Miller, S.C.; Kurz, W.; Harvey, D.

    1984-01-01

    Saccharomyces cerevisiae possesses a high-affinity estrogen binding protein and an endogenous ligand that displaces [3H]estradiol from both the yeast binding protein and mammalian estrogen receptors. Semipurified preparations of this ligand have been shown to exhibit estrogenic activity in mammalian systems. We now describe the purification procedure and ultimate identification of the estrogenic substance in extracts of S. cerevisiae as 17 beta-estradiol. Organic solvent extracts of commercia...

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

    OpenAIRE

    Jansen, M.L.A.; Winde, J.H. de; Pronk, J T

    2002-01-01

    When wild-type Saccharomyces cerevisiae strains pregrown in maltose-limited chemostat cultures were exposed 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 strain in which all members of the hexose transporter (H...

  8. Ethanol production from sugarbeet by Saccharomyces cerevisiae and Zymomonas mobilis strains

    Energy Technology Data Exchange (ETDEWEB)

    Nain, L.R.

    1985-01-01

    Examines the potential of sugar beet as a raw material for ethanol fermentation. The ethanol production from sugarbeet by 23 strains of Saccharomyces belonging to cerevisiae and uvarum species and four strains of Zymomonas mobilis was studied in batch fermentation. After screening, S. cerevisiae CDRI NTG was compared. The economics of ethanol production from sugar beet was calculated and the cost was found to be Rs. 2.50 per liter (excluding excise duty). (Refs. 187).

  9. Genomic Sequence Diversity and Population Structure of Saccharomyces cerevisiae Assessed by RAD-seq

    OpenAIRE

    Cromie, Gareth A.; Hyma, Katie E.; Ludlow, Catherine L.; Garmendia-torres, Cecilia; Gilbert, Teresa L.; May, Patrick; Huang, Angela A.; Dudley, Aime?e M.; Fay, Justin C.

    2013-01-01

    The budding yeast Saccharomyces cerevisiae is important for human food production and as a model organism for biological research. The genetic diversity contained in the global population of yeast strains represents a valuable resource for a number of fields, including genetics, bioengineering, and studies of evolution and population structure. Here, we apply a multiplexed, reduced genome sequencing strategy (known as RAD-seq) to genotype a large collection of S. cerevisiae ...

  10. Production and reutilization of an extracellular phosphatidylinositol catabolite, glycerophosphoinositol, by Saccharomyces cerevisiae.

    OpenAIRE

    Patton, J. L.; Pessoa-brandao, L.; Henry, S. A.

    1995-01-01

    Phosphatidylinositol catabolism in Saccharomyces cerevisiae is known to result in the formation of extracellular glycerophosphoinositol (GroPIns). We now report that S. cerevisiae not only produces but also reutilizes extracellular GroPIns and that these processes are regulated in response to inositol availability. A wild-type strain uniformly prelabeled with [3H] inositol displayed dramatically higher extracellular GroPIns levels when cultured in medium containing inositol than when cultured...

  11. Coordinate regulation of phospholipid biosynthesis in Saccharomyces cerevisiae: pleiotropically constitutive opi1 mutant.

    OpenAIRE

    Klig, L. S.; Homann, M J; Carman, G.M.; Henry, S A

    1985-01-01

    Phospholipid metabolism in the Saccharomyces cerevisiae opi1 mutant, which excretes inositol and is constitutive for the biosynthetic enzyme inositol-1-phosphate synthase (M. Greenberg, P. Goldwasser, and S. Henry, Mol. Gen. Genet. 186:157-163, 1982), was examined and compared to that of a wild-type strain. In wild-type S. cerevisiae, the phospholipid composition and the relative rates of synthesis of individual phospholipids change in response to the availability of exogenous supplies of sol...

  12. Analysis of the Metabolic Response of Saccharomyces Cerevisiae to DNA Damaging Agents

    OpenAIRE

    Rey, Simon Scheel

    2011-01-01

    Saccharomyces cerevisiae, commonly known as Baker?s yeast, is a eukaryotic model organism widely used in biotechnology research. Its genome has a high degree of similarity to humans, and research done on S. cerevisiae can give us a better understanding of the mechanisms involved and the cellular responses to anti-cancer drugs. Yeast is therefore usefool in increasing the effectiveness of anti-cancer drugs.The main goal for this master thesis was to investigate the metabolic response of S...

  13. Aminoacid metabolism and the production of wine aroma compounds by Saccharomyces cerevisiae

    OpenAIRE

    Arau?jo, Leandro Dias

    2012-01-01

    Os produtos do metabolismo de aminoácidos por leveduras Saccharomyces cerevisiae durante a fermentação alcoólica têm um impacto relevante no perfil sensorial de vinhos. Com o objetivo de estudar a influência da composição de aminoácidos do meio na formação da base do aroma típico de vinho, meios sintéticos contendo diferentes compostos nitrogenados foram fermentados por S. cerevisiae. Em um primeiro experimento, cinco meios de composições distintas quanto à fonte de azoto fora...

  14. Gene length and codon usage bias in Drosophila melanogaster, Saccharomyces cerevisiae and Escherichia coli.

    OpenAIRE

    Moriyama, E. N.; Powell, J. R.

    1998-01-01

    The relationship between gene length and synonymous codon usage bias was investigated in Drosophila melanogaster, Escherichia coli and Saccharomyces cerevisiae. Simulation studies indicate that the correlations observed in the three organisms are unlikely to be due to sampling errors or any potential bias in the methods used to measure codon usage bias. The correlation was significantly positive in E.coli genes, whereas negative correlations were obtained for D. melanogaster and S.cerevisiae ...

  15. Glutatation Transferasas de clase Omega en Saccharomyces cerevisiae: Estudio Bioquímico y Funcional

    OpenAIRE

    Barreto Parra, Lina Patricia

    2007-01-01

    Saccharomyces cerevisiae posseeix dues glutatió transferases (GST) anomenades Gtt1i Gtt2, amb capacitat de conjugar una molècula de glutatió amb el substrat estàndarCDNB. Aquests dos enzims no són clasificables dins de les classes convencionalsdescrites en base a l'estructura de les GST d'eucariotes superiors, encara que guardencerta similitud estructural amb els membres de la classe Zeta. En aquesta memòria esdescriu la caracterització de tres GST de classe Omega en S. cerevisiae anom...

  16. Study of genetic and phenotypic relationships in a Saccharomyces cerevisiae strain collection using computational approaches

    OpenAIRE

    Duarte, Ricardo Franco; Mendes, Ine?s; Umek, Lan; Neves, J. Drumonde; Zupan, Blaz; Schuller, Dorit

    2012-01-01

    Genome sequencing is essential to understand individual variation and to study the relationship between genotype and phenotype. Recently, large-scale sequencing projects of Saccharomyces cerevisiae revealed the existence of a few well defined lineages and some mosaics of that lineages, and suggested the occurrence of two domestication events during the history of association to human activities, one for sake strains and one for wine yeasts. Although the diversity of S. cerevisiae strains in w...

  17. Role of Nitrogen and Carbon Transport, Regulation, and Metabolism Genes for Saccharomyces cerevisiae Survival In Vivo†

    OpenAIRE

    Kingsbury, Joanne M.; Goldstein, Alan L.; Mccusker, John H.

    2006-01-01

    Saccharomyces cerevisiae is both an emerging opportunistic pathogen and a close relative of pathogenic Candida species. To better understand the ecology of fungal infection, we investigated the importance of pathways involved in uptake, metabolism, and biosynthesis of nitrogen and carbon compounds for survival of a clinical S. cerevisiae strain in a murine host. Potential nitrogen sources in vivo include ammonium, urea, and amino acids, while potential carbon sources include glucose, lactate,...

  18. Rapid Identification and Enumeration of Saccharomyces cerevisiae Cells in Wine by Real-Time PCR

    OpenAIRE

    Martorell, P.; Querol, A.; Ferna?ndez-espinar, M. T.

    2005-01-01

    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 designed for S. cerevisiae using the sequence information obtained from a cloned random amplified polymorphic DNA band that ...

  19. Identification and Characterization of a Novel Biotin Biosynthesis Gene in Saccharomyces cerevisiae

    OpenAIRE

    Wu, Hong; Ito, Kiyoshi; Shimoi, Hitoshi

    2005-01-01

    Yeast Saccharomyces cerevisiae cells generally cannot synthesize biotin, a vitamin required for many carboxylation reactions. Although sake yeasts, which are used for Japanese sake brewing, are classified as S. cerevisiae, they do not require biotin for their growth. In this study, we identified a novel open reading frame (ORF) in the genome of one strain of sake yeast that we speculated to be involved in biotin synthesis. Homologs of this gene are widely distributed in the genomes of sake ye...

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

    OpenAIRE

    Schuller, Dorit; Casal, Margarida

    2005-01-01

    During the last 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, continuously extended. The demand for suitable genetically modified (GM) S. cerevisiae strains for the biofuel, bakery and beverage industries or for the production of biotechnological products (e.g....

  1. Genomewide Expression Profiling of Cryptolepine-Induced Toxicity in Saccharomyces cerevisiae?

    OpenAIRE

    Rojas, Marta; Colin W. Wright; Piña, Benjamín; Portugal, José

    2008-01-01

    We have used the budding yeast Saccharomyces cerevisiae to identify genes that may confer sensitivity in vivo to the antimalarial and cytotoxic agent cryptolepine. Five S. cerevisiae strains, with different genetic backgrounds in cell permeability and DNA damage repair mechanisms, were exposed to several concentrations of cryptolepine. Cryptolepine showed a relatively mild toxicity for wild-type strains, which was augmented by either increasing cell permeability (Deltaerg6 or ISE2 strains) or...

  2. The Bioconversion of Red Ginseng Ethanol Extract into Compound K by Saccharomyces cerevisiae HJ-014

    OpenAIRE

    Choi, Hak Joo; Kim, Eun A.; Kim, Dong Hee; Shin, Kwang-soo

    2014-01-01

    A ?-glucosidase producing yeast strain was isolated from Korean traditional rice wine. Based on the sequence of the YCL008c gene and analysis of the fatty acid composition, the isolate was identified as Saccharomyces cerevisiae strain HJ-014. S. cerevisiae HJ-014 produced ginsenoside Rd, F2, and compound K from the ethanol extract of red ginseng. The production was increased by shaking culture, where the bioconversion efficiency was increased 2-fold compared to standing culture. The producti...

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

    OpenAIRE

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

    2012-01-01

    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 metaboli...

  4. Ergosterol production from molasses by genetically modified Saccharomyces cerevisiae.

    Science.gov (United States)

    He, Xiuping; Guo, Xuena; Liu, Nan; Zhang, Borun

    2007-05-01

    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) reductase and sterol acyltransferase in the yeast strain YEH56. The production of ergosterol by this recombinant strain using cane molasses (CM) as an inexpensive carbon source was investigated. An ergosterol content of 52.6 mg/g was obtained with 6.1 g/l of biomass from CM medium containing 60 g/l of total sugar in 30 h in shake flask. The ergosterol yield was enhanced through the increasing cell biomass by supplementation of urea to a concentration of 6 g/l in molasses medium. Fermentation was performed in 5-l bioreactor using the optimized molasses medium. In batch fermentation, the effect of agitation velocity on ergosterol production was examined. The highest ergosterol yield was obtained at 400 rpm that increased 60.4 mg/l in comparison with the shake flask culture. In fed-batch fermentation, yeast cells were cultivated, firstly, in the starting medium containing molasses with 20 g/l of total sugar, 1.68 g/l of phosphate acid, and 6 g/l of urea (pH 5.4) for 5 h, then molasses containing 350 g/l of total sugar was fed exponentially into the bioreactor to keep the ethanol level in the broth below 0.5%. After 40 h of cultivation, the ergosterol yield reached 1,707 mg/l, which was 3.1-fold of that in the batch fermentation. PMID:17225097

  5. IMMOBILIZATION OF Saccharomyces cerevisiae IN RICE HULLS FOR ETHANOL PRODUCTION

    Directory of Open Access Journals (Sweden)

    Don-Hee Park

    2010-11-01

    Full Text Available The whole cell immobilization in ethanol fermentation can be done by using natural carriers or through synthetic carriers. All of these methods have the same purpose of retaining high cell concentrations within a certain defined region of space which leads to higher ethanol productivity. Lignocellulosic plant substance represents one of highly potential sources in ethanol production. Some studies have found that cellulosic substances substances can also be used as a natural carrier in cell immobilization by re-circulating pre-culture medium into a reactor. In this experiment, rice hulls without any treatment were used to immobilize Saccharomyces cerevisiae through semi solid state incubation combined with re-circulating pre-culture medium. The scanning electron microscopy (SEM pictures of the carrier show that the yeast cells are absorbed and embedded to the rice hull pore. In liquid batch fermentation system with an initial sugar concentration of 50 g/L, nearly 100% total sugar was consumed after 48 hours. This resulted in an ethanol yield of 0.32 g ethanol/g glucose, which is 62.7% of the theoretical value. Ethanol productivity of 0.59 g/(L.h is 2.3 fold higher than that of free cells which is 0.26 g/(L.h. An effort to reuse the immobilized cells in liquid fermentation system showed poor results due to cell desorption in the first batch which led to high sugar concentration inhibitory effect in the second batch fermentation. This might be solved by using semi solid fermentation process in the future work.

  6. Genetic Basis for Saccharomyces cerevisiae Biofilm in Liquid Medium

    Science.gov (United States)

    Andersen, Kaj Scherz; Bojsen, Rasmus; Sørensen, Laura Gro Rejkjær; Nielsen, Martin Weiss; Lisby, Michael; Folkesson, Anders; Regenberg, Birgitte

    2014-01-01

    Biofilm-forming microorganisms switch between two forms: free-living planktonic and sessile multicellular. Sessile communities of yeast biofilms in liquid medium provide a primitive example of multicellularity and are clinically important because biofilms tend to have other growth characteristics than free-living cells. We investigated the genetic basis for yeast, Saccharomyces cerevisiae, biofilm on solid surfaces in liquid medium by screening a comprehensive deletion mutant collection in the ?1278b background and found 71 genes that were essential for biofilm development. Quantitative northern blots further revealed that AIM1, ASG1, AVT1, DRN1, ELP4, FLO8, FMP10, HMT1, KAR5, MIT1, MRPL32, MSS11, NCP1, NPR1, PEP5, PEX25, RIM8, RIM101, RGT1, SNF8, SPC2, STB6, STP22, TEC1, VID24, VPS20, VTC3, YBL029W, YBL029C-A, YFL054C, YGR161W-C, YIL014C-A, YIR024C, YKL151C, YNL200C, YOR034C-A, and YOR223W controlled biofilm through FLO11 induction. Almost all deletion mutants that were unable to form biofilms in liquid medium also lost the ability to form surface-spreading biofilm colonies (mats) on agar and 69% also lost the ability to grow invasively. The protein kinase A isoform Tpk3p functioned specifically in biofilm and mat formation. In a tpk3 mutant, transcription of FLO11 was induced three-fold compared with wild-type, but biofilm development and cell–cell adhesion was absent, suggesting that Tpk3p regulates FLO11 positive posttranscriptionally and negative transcriptionally. The study provides a resource of biofilm-influencing genes for additional research on biofilm development and suggests that the regulation of FLO11 is more complex than previously anticipated. PMID:25009170

  7. Isolation and characterization of acid phosphatase mutants in Saccharomyces cerevisiae.

    Science.gov (United States)

    To-E, A; Ueda, Y; Kakimoto, S I; Oshima, Y

    1973-02-01

    Saccharomyces cerevisiae strain H-42 seems to have two kinds of acid phosphatase: one which is constitutive and one which is repressible by inorganic phosphate. The constitutive enzyme was significantly unstable to heat inactivation, and its K(m) of 9.1 x 10(-4)m for p-nitrophenylphosphate was higher than that of the repressible enzyme (2.4 x 10(-4)m). The constitutive and the repressible acid phosphatases are specified by the phoC gene and by the phoB, phoD, or phoE gene, respectively. Results of tetrad analysis suggested that the phoC and phoE genes are linked to the lys2 locus on chromosome II. Since both repressible acid and alkaline phosphatases were affected simultaneously in the phoR, phoD, and phoS mutants, it was concluded that these enzymes were under the same regulatory mechanism or that they shared a common polypeptide. The phoR mutant produced acid phosphatase constitutively, and the phoR mutant allele was recessive to its wild-type counterpart. The phoS mutant showed a phenotype similar to that of a mutant defective in one of the phoB, phoD, or phoE genes. However, the results of genetic analysis of the phoS mutant clearly indicated that the phoS gene is not a structural gene for either of the repressible acid and alkaline phosphatases, but is a kind of regulatory gene. According to the proposed model, the phoS gene controls the expression of the phoR gene, and inorganic phosphate would act primarily as an inducer for the formation of the phoR product which represses phosphatase synthesis. PMID:4570606

  8. Metabolic engineering of Saccharomyces cerevisiae for optimizing 3HP production

    DEFF Research Database (Denmark)

    Jensen, Niels Bjerg; Maury, Jerome

    2012-01-01

    The finite nature of fossil resources and the negative influence of CO2 emissions on the global climate are key drivers in development of new biological processes. These are based on renewable resources such as sugar, starch, and biomass and aim at replacing chemical production from fossil fuels. Polyacrylates are a substantial part of the different plastic varieties found on the market. This kind of plastic is derived from acrylic acid, which is currently produced from propylene, a by-product of ethylene and gasoline production. Annually, more than one billion kilograms of acrylic acid is produced and the market for acrylate products exceeds USD 100 billion. As an alternative to oil and gas derived acrylic acid, 3-hydroxypropionic (3HP) acid produced from renewable sources is highly desired, because 3HP can easily be converted into acrylic acid. We are setting out to produce 3HP in yeast Saccharomyces cerevisiae. One main reason for selecting Baker's yeast as host organism is that yeast has a high tolerance towards low pH in comparison to bacteria, e.g. E. coli. Hence, it lowers the consumption of base for neutralization of growth media when compared to bacteria. The preferred engineered pathway towards 3HP has a substantial need for NADPH equivalents. Consequently, a yeast host with elevated NADPH availability is preferred. We will redirect several of the glycolysis steps in order to increase the NADPH generation per glucose molecule and thereby increase 3HP production. We believe this strain will be of high interest for other NADPH demanding biosynthetic routes.

  9. Predicting functional upstream open reading frames in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Kristiansson Erik

    2009-12-01

    Full Text Available Abstract Background Some upstream open reading frames (uORFs regulate gene expression (i.e., they are functional and can play key roles in keeping organisms healthy. However, how uORFs are involved in gene regulation is not yet fully understood. In order to get a complete view of how uORFs are involved in gene regulation, it is expected that a large number of experimentally verified functional uORFs are needed. Unfortunately, wet-experiments to verify that uORFs are functional are expensive. Results In this paper, a new computational approach to predicting functional uORFs in the yeast Saccharomyces cerevisiae is presented. Our approach is based on inductive logic programming and makes use of a novel combination of knowledge about biological conservation, Gene Ontology annotations and genes' responses to different conditions. Our method results in a set of simple and informative hypotheses with an estimated sensitivity of 76%. The hypotheses predict 301 further genes to have 398 novel functional uORFs. Three (RPC11, TPK1, and FOL1 of these 301 genes have been hypothesised, following wet-experiments, by a related study to have functional uORFs. A comparison with another related study suggests that eleven of the predicted functional uORFs from genes LDB17, HEM3, CIN8, BCK2, PMC1, FAS1, APP1, ACC1, CKA2, SUR1, and ATH1 are strong candidates for wet-lab experimental studies. Conclusions Learning based prediction of functional uORFs can be done with a high sensitivity. The predictions made in this study can serve as a list of candidates for subsequent wet-lab verification and might help to elucidate the regulatory roles of uORFs.

  10. Studies of anaerobic and aerobic glycolysis in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    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 experiments, which measured the concentration of [1-13C]glucose, whereas the distribution of end products was determined from the 13C and the 14C experiments. By combining these measurements the flows into the various pathways that contribute to glucose catabolism were estimated, and the effect of oxygen upon glucose catabolism was evaluated. From these measurements, the Pasteur quotient (PQ) for glucose catabolism was calculated to be 2.95 for acetate-grown cells and 1.89 for cells grown on glucose into saturation. The Warburg experiments provided an independent estimate of glucose catabolism. The PQ estimated from Warburg experiments was 2.9 for acetate-grown cells in excellent agreement with the labeled carbon experiments and 4.6 for cells grown into saturation, which did not agree. Possible explanations of these differences are discussed. From these data an estimate is obtained of the net flow through the Embden-Meyerhof-Parnas pathway. The backward flow through fructose-1,6-bisphosphatase (Fru-1,6-P2-ase) was calculated from the scrambling of the 13C label of [1-13C]the scrambling of the 13C label of [1-13C]glucose into the C1 and C6 positions of trehalose. Combining these data allowed us to calculate the net flux through phosphofructokinase (PFK). For acetate-grown cells we found that the relative flow through PFK is a factor of 1.7 faster anaerobically than aerobically

  11. Genetic Basis for Saccharomyces cerevisiae Biofilm in Liquid Medium

    DEFF Research Database (Denmark)

    Andersen, Kaj Scherz; Bojsen, Rasmus Kenneth

    2014-01-01

    Biofilm-forming microorganisms switch between two forms: free-living planktonic and sessile multicellular. Sessile communities of yeast biofilms in liquid medium provide a primitive example of multicellularity and are clinically important because biofilms tend to have other growth characteristics than free-living cells. We investigated the genetic basis for yeast, Saccharomyces cerevisiae, biofilm on solid surfaces in liquid medium by screening a comprehensive deletion mutant collection in the S1278b background and found 71 genes that were essential for biofilm development. Quantitative northern blots further revealed that AIM1, ASG1, AVT1, DRN1, ELP4, FLO8, FMP10, HMT1, KAR5, MIT1, MRPL32, MSS11, NCP1, NPR1, PEP5, PEX25, RIM8, RIM101, RGT1, SNF8, SPC2, STB6, STP22, TEC1, VID24, VPS20, VTC3, YBL029W, YBL029C-A, YFL054C, YGR161W-C, YIL014C-A, YIR024C, YKL151C, YNL200C, YOR034C-A, and YOR223W controlled biofilm through FLO11 induction. Almost all deletion mutants that were unable to form biofilms in liquid medium also lost the ability to form surface-spreading biofilm colonies (mats) on agar and 69% also lost the ability to grow invasively. The protein kinase A isoform Tpk3p functioned specifically in biofilm and mat formation. In a tpk3 mutant, transcription of FLO11 was induced three-fold compared with wild-type, but biofilm development and cell–cell adhesion was absent, suggesting that Tpk3p regulates FLO11 positive posttranscriptionally and negative transcriptionally. The study provides a resource of biofilm-influencing genes for additional research on biofilm development and suggests that the regulation of FLO11 is more complex than previously anticipated.

  12. Construction of lactose-consuming Saccharomyces cerevisiae for lactose fermentation into ethanol fuel.

    Science.gov (United States)

    Zou, Jing; Guo, Xuewu; Shen, Tong; Dong, Jian; Zhang, Cuiying; Xiao, Dongguang

    2013-04-01

    Two lactose-consuming diploid Saccharomyces cerevisiae strains, AY-51024A and AY-51024M, were constructed by expressing the LAC4 and LAC12 genes of Kluyveromyces marxianus in the host strain AY-5. In AY-51024A, both genes were targeted to the ATH1 and NTH1 gene-encoding regions to abolish the activity of acid/neutral trehalase. In AY-51024M, both genes were respectively integrated into the MIG1 and NTH1 gene-encoding regions to relieve glucose repression. Physiologic studies of the two transformants under anaerobic cultivations in glucose and galactose media indicated that the expression of both LAC genes did not physiologically burden the cells, except for AY-51024A in glucose medium. Galactose consumption was initiated at higher glucose concentrations in the MIG1 deletion strain AY-51024M than in the corresponding wild-type strain and AY-51024A, wherein galactose was consumed until glucose was completely depleted in the mixture. In lactose medium, the Sp. growth rates of AY-51024A and AY-51024M under anaerobic shake-flasks were 0.025 and 0.067 h(-1), respectively. The specific lactose uptake rate and ethanol production of AY-51024M were 2.50 g lactose g CDW(-1) h(-1) and 23.4 g l(-1), respectively, whereas those of AY-51024A were 0.98 g lactose g CDW(-1) h(-1) and 24.3 g lactose g CDW(-1) h(-1), respectively. In concentrated cheese whey powder solutions, AY-51024M produced 63.3 g l(-1) ethanol from approximately 150 g l(-1) initial lactose in 120 h, conversely, AY-51024A consumed 63.7 % of the initial lactose and produced 35.9 g l(-1) ethanol. Therefore, relieving glucose repression is an effective strategy for constructing lactose-consuming S. cerevisiae. PMID:23344501

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

    Directory of Open Access Journals (Sweden)

    de Winde Johannes H

    2008-02-01

    Full Text Available Abstract 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 relief of glucose limitation. Results The shift towards fully fermentative conditions caused a massive transcriptional reprogramming, where one third of all genes within the genome were transcribed differentially. The changes in transcript levels were mostly driven by relief from glucose-limitation. After an initial strong response to the addition of glucose, the expression profile of most transcriptionally regulated genes displayed a clear switch at 30 minutes. In this respect, a striking difference was observed between the transcript profiles of genes encoding ribosomal proteins and those encoding ribosomal biogenesis components. Not all regulated genes responded with this binary profile. A group of 87 genes showed a delayed and steady increase in expression that specifically responded to anaerobiosis. Conclusion Our study demonstrated that, despite the complexity of this multiple-input perturbation, the transcriptional responses could be categorized and biologically interpreted. By comparing this study with public datasets representing dynamic and steady conditions, 14 up-regulated and 11 down-regulated genes were determined to be anaerobic specific. Therefore, these can be seen as true "signature" transcripts for anaerobicity under dynamic as well as under steady state conditions.

  14. Acetaldehyde tolerance in Saccharomyces cerevisiae involves the pentose phosphate pathway and oleic acid biosynthesis.

    Science.gov (United States)

    Matsufuji, Yoshimi; Fujimura, Shuki; Ito, Takashi; Nishizawa, Makoto; Miyaji, Tatsuro; Nakagawa, Junichi; Ohyama, Tohru; Tomizuka, Noboru; Nakagawa, Tomoyuki

    2008-11-01

    To identify genes responsible for acetaldehyde tolerance, genome-wide screening was performed using a collection of haploid Saccharomyces cerevisiae strains deleted in single genes. The screen identified 49 genes whose deletion conferred acetaldehyde sensitivity, and these were termed the genes required for acetaldehyde tolerance. We focused on six of these genes required for acetaldehyde tolerance, ZWF1, GND1, RPE1, TKL1 and TAL1, which encode enzymes in the pentose phosphate pathway (PPP), and OAR1, which encodes for NADPH-dependent 3-oxoacyl-(acyl-carrier-protein) reductase. These genes were not only responsible for acetaldehyde tolerance but also turned out to be induced by acetaldehyde. Moreover, the content of oleic acid was remarkably increased in yeast cells under acetaldehyde stress, and supplementation of oleic acid into the media partially alleviated acetaldehyde stress-induced growth inhibition of strains disrupted in the genes required for acetaldehyde tolerance and OLE1. Taken together, our data suggest that the supply of NADPH and the process of fatty acid biosynthesis are the key factors in acetaldehyde tolerance in the yeast, and that oleic acid plays an important role in acetaldehyde tolerance. PMID:19061187

  15. Cloning of two cellobiohydrolase genes from Trichoderma viride and heterogeneous expression in yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Song, Jinzhu; Liu, Beidong; Liu, Zhihua; Yang, Qian

    2010-04-01

    Cellobiohydrolase genes cbhI and cbhII were isolated from Trichoderma viride AS3.3711 and T. viride CICC 13038, respectively, using RT-PCR technique. The cbhI gene from T. viride AS3.3711 contains 1,542 nucleotides and encodes a 514-amino acid protein with a molecular weight of approximately 53.96 kDa. The cbhII gene from T. viride CICC 13038 was 1,413 bp in length encoding 471 amino acid residues with a molecular weight of approximately 49.55 kDa. The CBHI protein showed high homology with enzymes belonging to glycoside hydrolase family 7 and CBHII is a member of Glycoside hydrolase family 6. CBHI and CBHII play a role in the conversion of cellulose to glucose by cutting the disaccharide cellobiose from the non-reducing end of the cellulose polymer chain. The two cellobiohydrolase (CBHI, CBHII) genes were successfully expressed in Saccharomyces cerevisiae H158. Maximal activities of transformants Sc-cbhI and Sc-cbhII were 0.03 and 0.089 units ml(-1) under galactose induction, respectively. The optimal temperatures of the recombinant enzymes (CBHI, CBHII) were 60 and 70 degrees C, respectively. The optimal pHs of recombinant enzymes CBHI and CBHII were at pH 5.8 and 5.0, respectively. PMID:19669931

  16. Accumulation and chemical states of radiocesium by fungus Saccharomyces cerevisiae

    Science.gov (United States)

    Ohnuki, Toshihiko; Sakamoto, Fuminori; Kozai, Naofumi; Yamasaki, Shinya; Yu, Qianqian

    2014-05-01

    After accident of Fukushima Daiichi Nuclear Power Plant, the fall-out radiocesium was deposited on the ground. Filamentous fungus is known to accumulate radiocesium in environment, even though many minerals are involved in soil. These facts suggest that fungus affect the migration behavior of radiocesium in the environment. However, accumulation mechanism of radiocesium by fungus is not understood. In the present study, accumulation and chemical states change of Cs by unicellular fungus of Saccharomyces cerevisiae have been studied to elucidate the role of microorganisms in the migration of radiocesium in the environment. Two different experimental conditions were employed; one is the accumulation experiments of radiocesium by S. cerevisiae from the agar medium containing 137Cs and a mineral of zeolite, vermiculite, smectite, mica, or illite. The other is the experiments using stable cesium to examine the chemical states change of Cs. In the former experiment, the cells were grown on membrane filter of 0.45 ?m installed on the agar medium. After the grown cells were weighed, radioactivity in the cells was measured by an autoradiography technique. The mineral weight contents were changed from 0.1% to 1% of the medium. In the latter experiment, the cells were grown in the medium containing stable Cs between 1 mM and 10mM. The Cs accumulated cells were analyzed by SEM-EDS and EXAFS. The adsorption experiments of cesium by the cells under resting condition were also conducted to test the effect of cells metabolic activity. Without mineral in the medium, cells of S. cerevisiae accumulated 1.5x103 Bq/g from the medium containing 137Cs of 2.6x102 Bq/g. When mineral was added in the medium, concentration of 137Cs in the cells decreased. The concentration of 137Cs in the cells from the medium containing different minerals were in the following order; smectite, illite, mica > vermiculite > zeolite. This order was nearly the same as the inverse of distribution coefficient of mineral for 137Cs in the medium solution. The concentration of 137Cs in the cells lowered in the medium containing higher mineral content. These results indicate that radiocesium was competively accumulated in the cells with minerals in the soil. Higher concentration of stable Cs was accumulated in the cells in the metabolically active condition than in the resting cells condition. XAFS analyses showed that the k3-weighted extended-XAFS functions and the radial structural function of Cs accumulated by the cells in the metabolically active condition were similar to those in the resting condition, indicating that chemical states of the accumulated Cs were nearly the same between both conditions. These results indicate that the fungus accumulates radiocesium by competitively with minerals in the soils, and performs higher retardation of the migration of Cs in the metabolically active condition than the resting one. A part of this study is the results of "Multidisciplinary investigation on radiocesium fate and transport for safety assessment for interim storage and disposal of heterogeneous waste" carried out under the Initiatives for Atomic Energy Basic and Generic Strategic Research by the Ministry of Education, Culture, Sports, Science and Technology of Japan.

  17. [Construction of high sulphite-producing industrial strain of Saccharomyces cerevisiae].

    Science.gov (United States)

    Qu, Na; He, Xiu-ping; Guo, Xue-na; Liu, Nan; Zhang, Bo-run

    2006-02-01

    In the process of beer storage and transportation, off-flavor can be produced for oxidation of beer. Sulphite is important for stabilizing the beer flavor because of its antioxidant activity. However, the low level of sulphite synthesized by the brewing yeast is not enough to stabilize beer flavor. Three enzymes involve sulphite biosynthesis in yeast. One of them, APS kinase (encoded by MET14) plays important role in the process of sulphite formation. In order to construct high sulphite-producing brewing yeast strain for beer production, MET14 gene was cloned and overexpressed in industrial strain of Saccharomyces cerevisiae. Primer 1 (5'-TGTGAATTCCTGTACACCAATGGCTACT-3', EcoR I) and primer 2 (5'-TATAAGCTTGATGA GGTGGATGAAGACG-3', HindIII) were designed according to the MET14 sequence in GenBank. A 1.1kb DNA fragment containing the open reading frame and terminator of MET14 gene was amplified from Saccharomyces cerevisiae YSF-5 by PCR, and inserted into YEp352 to generate recombinant plasmid pMET14. To express MET14 gene properly in S. cerevisiae, the recombinant expression plasmids pPM with URA3 gene as the selection marker and pCPM with URA3 gene and copper resistance gene as the selection marker for yeast transformation were constructed. In plasmid pPM, the PGK1 promoter from plasmid pVC727 was fused with the MET14 gene from pMET14, and the expression cassette was inserted into the plasmid YEp352. The dominant selection marker, copper-resistance gene expression cassette CUP1-MTI was inserted in plasmid pPM to result in pCPM. Restriction enzyme analysis showed that plasmids pPM and pCPM were constructed correctly. The laboratory strain of S. cerevisiae YS58 with ura3, trp1, leu2, his4 auxotroph was transformed with plasmid pPM. Yeast transformants were screened on synthetic minimal medium (SD) containing leucine, histidine and tryptophan. The sulphite production of the transformants carrying pPM was 2 fold of that in the control strain YS58, which showed that the MET14 gene on plasmid pPM was expressed functionally in YS58. The industrial brewing yeast strain YSF-38 was transformed with the plasmid pCPM and yeast transformants were selected on YEPD medium containing 4mmol/L copper sulphate. The recombinant strain carrying pCPM showed a 3.2-fold increase in sulphite production when compared to the host strain YSF-38 under laboratory culture conditions. Flask fermentation under brewing-like conditions was performed in Tsingtao Beer Brewery. The sulphite production of the recombinant strain began to be higher than that of the host strain YSF-38 at the fourth day and reached the maximum at the eighth day. At the end of fermentation, the sulphite produced by recombinant strain is 1.4 fold of that in the host strain. The overexpression of MET14 gene in both laboratory and industrial strains of S. cerevisiae increases the sulphite formation. It is the first time to construct high sulphite-producing industrial strain by functional expression of MET14 in S. cerevisiae. Such study provides the foundation for construction of an excellent brewing yeast strain that can produce proper sulphite and can be used in commercial beer production. PMID:16579462

  18. Saccharomyces kudriavzevii and Saccharomyces uvarum differ from Saccharomyces cerevisiae during the production of aroma-active higher alcohols and acetate esters using their amino acidic precursors.

    Science.gov (United States)

    Stribny, Jiri; Gamero, Amparo; Pérez-Torrado, Roberto; Querol, Amparo

    2015-07-16

    Higher alcohols and acetate esters are important flavour and aroma components in the food industry. In alcoholic beverages these compounds are produced by yeast during fermentation. Although Saccharomyces cerevisiae is one of the most extensively used species, other species of the Saccharomyces genus have become common in fermentation processes. This study analyses and compares the production of higher alcohols and acetate esters from their amino acidic precursors in three Saccharomyces species: Saccharomyces kudriavzevii, Saccharomyces uvarum and S. cerevisiae. The global volatile compound analysis revealed that S. kudriavzevii produced large amounts of higher alcohols, whereas S. uvarum excelled in the production of acetate esters. Particularly from phenylalanine, S. uvarum produced the largest amounts of 2-phenylethyl acetate, while S. kudriavzevii obtained the greatest 2-phenylethanol formation from this precursor. The present data indicate differences in the amino acid metabolism and subsequent production of flavour-active higher alcohols and acetate esters among the closely related Saccharomyces species. This knowledge will prove useful for developing new enhanced processes in fragrance, flavour, and food industries. PMID:25886016

  19. Viabilidade celular de Saccharomyces cerevisiae cultivada em associação com bactérias contaminantes da fermentação alcoólica / Cellular viability of Saccharomyces cerevisiae cultivated in association with contaminant bacteria of alcoholic fermentation

    Scientific Electronic Library Online (English)

    Thais de Paula, Nobre; Jorge, Horii; André Ricardo, Alcarde.

    2007-03-01

    Full Text Available O objetivo deste trabalho foi estudar a influência de bactérias dos gêneros Bacillus e Lactobacillus, bem como de seus produtos metabólicos, na redução da viabilidade celular de leveduras Saccharomyces cerevisiae. As bactérias Bacillus subtilis, Bacillus coagulans, Bacillus stearothermophilus, Lacto [...] bacillus fermentum e Lactobacillus plantarum foram cultivadas em associação com a levedura S. cerevisiae (cepa Y-904) por 72 horas a 32 °C, sob agitação. A viabilidade celular, a taxa de brotamento e a população de células de S. cerevisiae e a acidez total, a acidez volátil e o pH dos meios de cultivos foram determinados às 0, 24, 48 e 72 horas do cultivo misto. As culturas de bactérias foram tratadas através do calor, de agente antimicrobiano e de irradiação. Os resultados mostraram que apenas os meios de cultivo mais acidificados, contaminados com as bactérias ativas L. fermentum e B. subtilis, provocaram redução na viabilidade celular de S. cerevisiae. Excetuando a bactéria B. subtilis tratada com radiação gama, as demais bactérias tratadas pelos diferentes processos (calor, irradiação e antimicrobiano) não causaram diminuição da viabilidade celular e da população de S. cerevisiae, indicando que a presença isolada dos metabólitos celulares dessas bactérias não foi suficiente para reduzir a porcentagem de células vivas de S. cerevisiae. Abstract in english The aim of this project was to study the influence of the bacteria Bacillus and Lactobacillus, as well as their metabolic products to decrease the cellular viability of the yeast Saccharomyces cerevisiae. The bacteria Bacillus subtilis, Bacillus coagulans, Bacillus stearothermophilus, Lactobacillus [...] fermentum and Lactobacillus plantarum were cultivated in association with yeast S. cerevisiae (strain Y-904) for 72 hours at 32 ºC under agitation. The cellular viability, budding rate and population of S. Cerevisiae and the total acidity, volatile acidity and pH of culture medium were determined at 0, 24, 48 and 72 hours of incubation of the mixed culture. The bacteria cultures were treated by heat sterilization, antibacterial agent and irradiation. The results showed that only the more acidified culture medium, contaminated with active bacteria L. fermentum and B. subtilis, caused a reduction in the yeast cellular viability. Except for the bacteria B. subtilis treated for radiation, the other bacteria treated by the different procedures (heat, radiation and antibacterial) did not cause a reduction in the cellular viability of S. cerevisiae, indicating that the isolated presence of the cellular metabolic of these bacteria was not enough to reduce the percentage of the living yeast cells.

  20. Viabilidade celular de Saccharomyces cerevisiae cultivada em associação com bactérias contaminantes da fermentação alcoólica Cellular viability of Saccharomyces cerevisiae cultivated in association with contaminant bacteria of alcoholic fermentation

    Directory of Open Access Journals (Sweden)

    Thais de Paula Nobre

    2007-03-01

    Full Text Available O objetivo deste trabalho foi estudar a influência de bactérias dos gêneros Bacillus e Lactobacillus, bem como de seus produtos metabólicos, na redução da viabilidade celular de leveduras Saccharomyces cerevisiae. As bactérias Bacillus subtilis, Bacillus coagulans, Bacillus stearothermophilus, Lactobacillus fermentum e Lactobacillus plantarum foram cultivadas em associação com a levedura S. cerevisiae (cepa Y-904 por 72 horas a 32 °C, sob agitação. A viabilidade celular, a taxa de brotamento e a população de células de S. cerevisiae e a acidez total, a acidez volátil e o pH dos meios de cultivos foram determinados às 0, 24, 48 e 72 horas do cultivo misto. As culturas de bactérias foram tratadas através do calor, de agente antimicrobiano e de irradiação. Os resultados mostraram que apenas os meios de cultivo mais acidificados, contaminados com as bactérias ativas L. fermentum e B. subtilis, provocaram redução na viabilidade celular de S. cerevisiae. Excetuando a bactéria B. subtilis tratada com radiação gama, as demais bactérias tratadas pelos diferentes processos (calor, irradiação e antimicrobiano não causaram diminuição da viabilidade celular e da população de S. cerevisiae, indicando que a presença isolada dos metabólitos celulares dessas bactérias não foi suficiente para reduzir a porcentagem de células vivas de S. cerevisiae.The aim of this project was to study the influence of the bacteria Bacillus and Lactobacillus, as well as their metabolic products to decrease the cellular viability of the yeast Saccharomyces cerevisiae. The bacteria Bacillus subtilis, Bacillus coagulans, Bacillus stearothermophilus, Lactobacillus fermentum and Lactobacillus plantarum were cultivated in association with yeast S. cerevisiae (strain Y-904 for 72 hours at 32 ºC under agitation. The cellular viability, budding rate and population of S. Cerevisiae and the total acidity, volatile acidity and pH of culture medium were determined at 0, 24, 48 and 72 hours of incubation of the mixed culture. The bacteria cultures were treated by heat sterilization, antibacterial agent and irradiation. The results showed that only the more acidified culture medium, contaminated with active bacteria L. fermentum and B. subtilis, caused a reduction in the yeast cellular viability. Except for the bacteria B. subtilis treated for radiation, the other bacteria treated by the different procedures (heat, radiation and antibacterial did not cause a reduction in the cellular viability of S. cerevisiae, indicating that the isolated presence of the cellular metabolic of these bacteria was not enough to reduce the percentage of the living yeast cells.

  1. Isolation and characterization of Saccharomyces cerevisiae strains of winery interest Isolamento e caracterização de cepas de Saccharomyces cerevisiae de interesse em produção de vinho

    Directory of Open Access Journals (Sweden)

    Thais M. Guimarães

    2006-03-01

    Full Text Available Despite the availability of several Saccharomyces cerevisiae commercial strains intended for wine production, strains isolated from winery regions are usually more adapted to their own climatic conditions, grapes and also partially responsible for particular characteristics that frequently identify specific wines and regions. Thus the microbiota of an important winery region (Colombo was studied in order to isolate and characterize S. cerevisiae strains that could be used on wine production. From 61 yeasts isolated, 14 were identified as S. cerevisiae. Some of them showed fermentative characteristics even better than commercial strains indicating that they could be applied on wine production in order to increase the quality and assure the particular wine characteristics of that region.

  2. Effects of low-frequency magnetic fields on the viability of yeast Saccharomyces cerevisiae.

    Czech Academy of Sciences Publication Activity Database

    Novák, Jan; Strašák, Lud?k; Fojt, Lukáš; Slaninová, I.; Vetterl, Vladimír

    2007-01-01

    Ro?. 70, ?. 1 (2007), s. 115-121. ISSN 1567-5394 R&D Projects: GA AV ?R(CZ) IAA4004404; GA AV ?R(CZ) IBS5004107 Institutional research plan: CEZ:AV0Z50040702 Keywords : low-frequency electromagnetic field * yeast * Saccharomyces cerevisiae Subject RIV: BO - Biophysics Impact factor: 2.992, year: 2007

  3. Growth-rate regulated genes have profound impact on interpretation of transcriptome profiling in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Regenberg, Birgitte; Grotkjaer, Thomas

    2006-01-01

    Growth rate is central to the development of cells in all organisms. However, little is known about the impact of changing growth rates. We used continuous cultures to control growth rate and studied the transcriptional program of the model eukaryote Saccharomyces cerevisiae, with generation times varying between 2 and 35 hours.

  4. Global LC/MS Metabolomics Profiling of Calcium Stressed and Immunosuppressant Drug Treated Saccharomyces cerevisiae

    OpenAIRE

    Stefan Jenkins; Fischer, Steven M.; Lily Chen; Sana, Theodore R.

    2013-01-01

    Previous studies have shown that calcium stressed Saccharomyces cerevisiae, challenged with immunosuppressant drugs FK506 and Cyclosporin A, responds with comprehensive gene expression changes and attenuation of the generalized calcium stress response. Here, we describe a global metabolomics workflow for investigating the utility of tracking corresponding phenotypic changes. This was achieved by efficiently analyzing relative abundance differences between intracellular metabolite pools from w...

  5. Induction of mutations by photodynamic action of thiopyronine in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    The induction of cytoplasmic and nuclear mutations by the photodynamic action of thiopyronine is demonstrated in a haploid strain of Saccharomyces cerevisiae that has been isolated as a photodynamic sensitive mutant. No significant increase in corresponding mutation frequensies could be observed in a strain resistant to photodynamic inactivation by thiopyronine. (orig.)

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

    Science.gov (United States)

    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...

  7. Engineering Saccharomyces cerevisiae for consolidated bioprocessing in starch and biomass conversion

    Science.gov (United States)

    The conversion of starch or biomass to biofuel is a two-stage process involving enzymatic treatment, followed by yeast fermentation. An alternative route would be to consolidate the process by engineering Saccharomyces cerevisiae capable of both saccharification and fermentation. An approach was d...

  8. Implementation of Sliding Mode Controller with Boundary Layer for Saccharomyces cerevisiae Fed-batch Cultivation

    Directory of Open Access Journals (Sweden)

    Stoyan Tzonkov

    2005-04-01

    Full Text Available An implementation of sliding mode control for yeast fed-batch cultivation is presented in this paper. Developed controller has been implemented on two real fed-batch cultivations of Saccharomyces cerevisiae. The controller successfully stabilizes the process and shows a very good performance at high input disturbances.

  9. Investigation of the effect of water exposed to nonequilibrium contact plasma onto saccharomyces cerevisiae yeast

    Directory of Open Access Journals (Sweden)

    S. Mykolenko

    2015-05-01

    Full Text Available Introduction. Additional treatment of water by nonequilibrium contact plasma allows improving consumer characteristics of bakery goods considerably. Determination of the effect of plasma-chemically activated water on morphological, cultural and physiological properties of Saccharomyces cerevisiae yeast is important from the technological point of view. Materials and Methods. Experimental investigations were carried out in the conditions of bacteriological laboratory by seeding the culture of yeasts of ?? “Lvivski” and “Kryvorizki” on Sabouraud dense liquid nutrient media. The quantity of viable cells of microorganisms was determined by the method of Gould sector seeds. Morphology of the yeast was investigated by phase-contrast microscopy. Biotechnological properties of yeasts were determined on Giss media. Results. The paper establishes the effect of water exposed to nonequilibrium contact plasma on the sensitivity of Saccharomyces cerevisiae and shows absence of suppressive action of treated water with regard to cultural properties of microorganisms. The experiments prove that with the use of plasma-chemically activated water morphological characteristics and biochemical properties of bakery yeasts produced by Lviv and Kryvyi Rig yeast plants are preserved. Culturing of Saccharomyces cerevisiae yeast on the nutrient media prepared with the use of water exposed to nonequilibrium contact plasm resulted in 6,5–15 times’ increase in quantity of viable microorganisms compared with the control on the mains drinking water. Conclusions. Physiological properties of Saccharomyces cerevisiae yeast improved owing to use water exposed to nonequilibrium contact plasma. Results of investigations are recommended for using in yeast production and bread making.

  10. Aged mother cells of Saccharomyces cerevisiae show markers of oxidative stress and apoptosis.

    Czech Academy of Sciences Publication Activity Database

    Laun, P.; Pichová, Alena; Madeo, F.; Fuchs, J.; Ellinger, A.; Kohlwein, S.; Dawes, I.; Fröhlich, K. U.; Breitenbach, M.

    2001-01-01

    Ro?. 39, ?. 5 (2001), s. 1166-1173. ISSN 0950-382X R&D Projects: GA ?R GA204/97/0541 Institutional research plan: CEZ:AV0Z5020903 Keywords : Saccharomyces cerevisiae * genetic changes Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 6.398, year: 2001

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

    Science.gov (United States)

    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...

  12. Saccharomyces cerevisiae Mixed Culture of Blackberry (Rubus ulmifolius L.) Juice: Synergism in the Aroma Compounds Production

    OpenAIRE

    Bautista-Rosales, Pedro Ulises; Ragazzo-Sánchez, Juan Arturo; Ruiz-Montañez, Gabriela; Ortiz-Basurto, Rosa Isela; Luna-Solano, Guadalupe; Calderón-Santoyo, Montserrat

    2014-01-01

    Blackberry (Rubus sp.) juice was fermented using four different strains of Saccharomyces cerevisiae (Vitilevure-CM4457, Enoferm-T306, ICV-K1, and Greroche Rhona-L3574) recognized because of their use in the wine industry. A medium alcoholic graduation spirit (

  13. Saccharomyces cerevisiae BY4741 and W303-1A laboratory strains differ in salt tolerance.

    Czech Academy of Sciences Publication Activity Database

    Petrezsélyová, Silvia; Zahrádka, Jaromír; Sychrová, Hana

    2010-01-01

    Ro?. 114, 2-3 (2010), s. 144-150. ISSN 1878-6146 R&D Projects: GA MŠk(CZ) LC531; GA ?R(CZ) GA204/08/0354 Institutional research plan: CEZ:AV0Z50110509 Keywords : Saccharomyces cerevisiae * salt tolerance * potassium homeostasis Subject RIV: EB - Genetics ; Molecular Biology

  14. The uptake of different iron salts by the yeast Saccharomyces cerevisiae

    Scientific Electronic Library Online (English)

    Fernanda, Gaensly; Geraldo, Picheth; Debora, Brand; Tania M.B., Bonfim.

    2014-06-01

    Full Text Available Yeasts can be enriched with microelements, including iron; however, special physicochemical conditions are required to formulate a culture media that promotes both yeast growth and iron uptake. Different iron sources do not affect biomass formation; however, considering efficacy, cost, stability, an [...] d compatibility with Saccharomyces cerevisiae metabolism, ferrous sulphate is recommended.

  15. Invert sugar formation with Saccharomyces cerevisiae cells encapsulated in magnetically responsive alginate microparticles.

    Czech Academy of Sciences Publication Activity Database

    Šafa?ík, Ivo; Mad?rová, Zde?ka; Šafa?íková, Miroslava

    2009-01-01

    Ro?. 321, - (2009), s. 1478-1481. ISSN 0304-8853 R&D Projects: GA MPO 2A-1TP1/094; GA MŠk(CZ) OC 157 Institutional research plan: CEZ:AV0Z60870520 Keywords : magnetic alginate microbeads * Saccharomyces cerevisiae * invertase Subject RIV: EI - Biotechnology ; Bionics Impact factor: 1.204, year: 2009

  16. Screening and identification of respiration deficiency mutants of yeasts (Saccharomyces Cerevisiae) induced by heavy ion irradiation

    International Nuclear Information System (INIS)

    A screen of respiration deficiency mutants of Saccharomyces Cerevisiae induced by 5.19 MeV/u 22Ne5- ion irradiation is studied. Some respiration deficiency mutants, which are white colony phenotype in the selective culture of TTC medium, are obtained. The mutants are effectively identified by means of a new and simplified restriction analysis method. (authors)

  17. Bioaccumulation of uranium from waste water using different strains of Saccharomyces cerevisiae.

    Czech Academy of Sciences Publication Activity Database

    Tykva, Richard; Novák, Jaroslav; Podracká, Eva; Popa, K.

    2009-01-01

    Ro?. 54, ?. 2 (2009), s. 143-148. ISSN 0029-5922 Institutional research plan: CEZ:AV0Z40550506 Keywords : bioaccumulation * uranium * Saccharomyces cerevisiae * competitive ions Subject RIV: CH - Nuclear ; Quantum Chemistry Impact factor: 0.159, year: 2009

  18. High-efficiency transformation of Saccharomyces cerevisiae cells by bacterial minicell protoplast fusion.

    OpenAIRE

    Gyuris, J; Duda, E G

    1986-01-01

    After a new transformation procedure, 10% of Saccharomyces cerevisiae cells were found to contain transforming DNA sequences. We used direct transfer of plasmid molecules by fusing bacterial minicell protoplasts to yeast protoplasts. Since the procedure significantly reduces the toxic effect of procaryotic protoplasm on the eucaryotic organism, it might be generally applicable in other systems in which transformation is inefficient or impossible.

  19. Mixing of vineyard and oak-tree ecotypes of Saccharomyces cerevisiae in North American vineyards.

    Science.gov (United States)

    Hyma, Katie E; Fay, Justin C

    2013-06-01

    Humans have had a significant impact on the distribution and abundance of Saccharomyces cerevisiae through its widespread use in beer, bread and wine production. Yet, similar to other Saccharomyces species, S. cerevisiae has also been isolated from habitats unrelated to fermentations. Strains of S. cerevisiae isolated from grapes, wine must and vineyards worldwide are genetically differentiated from strains isolated from oak-tree bark, exudate and associated soil in North America. However, the causes and consequences of this differentiation have not yet been resolved. Historical differentiation of these two groups may have been influenced by geographic, ecological or human-associated barriers to gene flow. Here, we make use of the relatively recent establishment of vineyards across North America to identify and characterize any active barriers to gene flow between these two groups. We examined S. cerevisiae strains isolated from grapes and oak trees within three North American vineyards and compared them to those isolated from oak trees outside of vineyards. Within vineyards, we found evidence of migration between grapes and oak trees and potential gene flow between the divergent oak-tree and vineyard groups. Yet, we found no vineyard genotypes on oak trees outside of vineyards. In contrast, Saccharomyces paradoxus isolated from the same sources showed population structure characterized by isolation by distance. The apparent absence of ecological or genetic barriers between sympatric vineyard and oak-tree populations of S. cerevisiae implies that vineyards play an important role in the mixing between these two groups. PMID:23286354

  20. Heterologous expression and characterization of bacterial 2-C-methyl-d-erythritol-4-phosphate pathway in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Carlsen, Simon; Ajikumar, Parayil Kumaran

    2013-01-01

    Transfer of a biosynthetic pathway between evolutionary distant organisms can create a metabolic shunt capable of bypassing the native regulation of the host organism, hereby improving the production of secondary metabolite precursor molecules for important natural products. Here, we report the engineering of Escherichia coli genes encoding the 2-C-methyl-d-erythritol-4-phosphate (MEP) pathway into the genome of Saccharomyces cerevisiae and the characterization of intermediate metabolites synthesized by the MEP pathway in yeast. Our UPLC-MS analysis of the MEP pathway metabolites from engineered yeast showed that the pathway is active until the synthesis of 2-C-methyl-d-erythritol-2,4-cyclodiphosphate, but appears to lack functionality of the last two steps of the MEP pathway, catalyzed by the [4Fe–4S] iron sulfur cluster proteins encoded by ispG and ispH. In order to functionalize the last two steps of the MEP pathway, we co-expressed the genes for the E. coli iron sulfur cluster (ISC) assembly machinery. Bydeleting ERG13, thereby incapacitating the mevalonate pathway, in conjunction with labeling experiments with U–13C6 glucose and growth experiments, we found that the ISC assembly machinery was unable to functionalize ispG and ispH. However, we have found that leuC and leuD, encoding the heterodimeric iron–sulfur cluster protein, isopropylmalate isomerase, can complement the S. cerevisiae leu1 auxotrophy. To our knowledge, this is the first time a bacterial iron–sulfur cluster protein has been functionally expressed in the cytosol of S. cerevisiae under aerobic conditions and shows that S. cerevisiae has the capability to functionally express at least some bacterial iron–sulfur cluster proteins in its cytosol.

  1. Crystal structure of Saccharomyces cerevisiae 6-phosphogluconate dehydrogenase Gnd1

    Directory of Open Access Journals (Sweden)

    Zhou Cong-Zhao

    2007-06-01

    Full Text Available Abstract Background As the third enzyme of the pentose phosphate pathway, 6-phosphogluconate dehydrogenase (6PGDH is the main generator of cellular NADPH. Both thioredoxin reductase and glutathione reductase require NADPH as the electron donor to reduce oxidized thioredoxin or glutathione (GSSG. Since thioredoxin and GSH are important antioxidants, it is not surprising that 6PGDH plays a critical role in protecting cells from oxidative stress. Furthermore the activity of 6PGDH is associated with several human disorders including cancer and Alzheimer's disease. The 3D structural investigation would be very valuable in designing small molecules that target this enzyme for potential therapeutic applications. Results The crystal structure of 6-phosphogluconate dehydrogenase (6PGDH/Gnd1 from Saccharomyces cerevisiae has been determined at 2.37 Å resolution by molecular replacement. The overall structure of Gnd1 is a homodimer with three domains for each monomer, a Rossmann fold NADP+ binding domain, an all-? helical domain contributing the majority to hydrophobic interaction between the two subunits and a small C-terminal domain penetrating the other subunit. In addition, two citrate molecules occupied the 6PG binding pocket of each monomer. The intact Gnd1 had a Km of 50 ± 9 ?M for 6-phosphogluconate and of 35 ± 6 ?M for NADP+ at pH 7.5. But the truncated mutants without the C-terminal 35, 39 or 53 residues of Gnd1 completely lost their 6PGDH activity, despite remaining the homodimer in solution. Conclusion The overall tertiary structure of Gnd1 is similar to those of 6PGDH from other species. The substrate and coenzyme binding sites are well conserved, either from the primary sequence alignment, or from the 3D structural superposition. Enzymatic activity assays suggest a sequential mechanism of catalysis, which is in agreement with previous studies. The C-terminal domain of Gnd1 functions as a hook to further tighten the dimer, but it is not necessary for the dimerization. This domain also works as a lid on the substrate binding pocket to control the binding of substrate and the release of product, so it is indispensable for the 6PGDH activity. Moreover, the co-crystallized citrate molecules, which mimic the binding mode of the substrate 6-phosphogluconate, provided us a novel strategy to design the 6PDGH inhibitors.

  2. "Ant" and "grasshopper" life-history strategies in Saccharomyces cerevisiae.

    Science.gov (United States)

    Spor, Aymé; Wang, Shaoxiao; Dillmann, Christine; de Vienne, Dominique; Sicard, Delphine

    2008-01-01

    From the evolutionary and ecological points of view, it is essential to distinguish between the genetic and environmental components of the variability of life-history traits and of their trade-offs. Among the factors affecting this variability, the resource uptake rate deserves particular attention, because it depends on both the environment and the genetic background of the individuals. In order to unravel the bases of the life-history strategies in yeast, we grew a collection of twelve strains of Saccharomyces cerevisiae from different industrial and geographical origins in three culture media differing for their glucose content. Using a population dynamics model to fit the change of population size over time, we estimated the intrinsic growth rate (r), the carrying capacity (K), the mean cell size and the glucose consumption rate per cell. The life-history traits, as well as the glucose consumption rate, displayed large genetic and plastic variability and genetic-by-environment interactions. Within each medium, growth rate and carrying capacity were not correlated, but a marked trade-off between these traits was observed over the media, with high K and low r in the glucose rich medium and low K and high r in the other media. The cell size was tightly negatively correlated to carrying capacity in all conditions. The resource consumption rate appeared to be a clear-cut determinant of both the carrying capacity and the cell size in all media, since it accounted for 37% to 84% of the variation of those traits. In a given medium, the strains that consume glucose at high rate have large cell size and low carrying capacity, while the strains that consume glucose at low rate have small cell size but high carrying capacity. These two contrasted behaviors may be metaphorically defined as "ant" and "grasshopper" strategies of resource utilization. Interestingly, a strain may be "ant" in one medium and "grasshopper" in another. These life-history strategies are discussed with regards to yeast physiology, and in an evolutionary perspective. PMID:18270570

  3. Increased isobutanol production in Saccharomyces cerevisiae by overexpression of genes in valine metabolism

    Directory of Open Access Journals (Sweden)

    Karhumaa Kaisa

    2011-07-01

    Full Text Available Abstract Background Isobutanol can be a better biofuel than ethanol due to its higher energy density and lower hygroscopicity. Furthermore, the branched-chain structure of isobutanol gives a higher octane number than the isomeric n-butanol. Saccharomyces cerevisiae was chosen as the production host because of its relative tolerance to alcohols, robustness in industrial fermentations, and the possibility for future combination of isobutanol production with fermentation of lignocellulosic materials. Results The yield of isobutanol was improved from 0.16 to 0.97 mg per g glucose by simultaneous overexpression of biosynthetic genes ILV2, ILV3, and ILV5 in valine metabolism in anaerobic fermentation of glucose in mineral medium in S. cerevisiae. Isobutanol yield was further improved by twofold by the additional overexpression of BAT2, encoding the cytoplasmic branched-chain amino-acid aminotransferase. Overexpression of ILV6, encoding the regulatory subunit of Ilv2, in the ILV2 ILV3 ILV5 overexpression strain decreased isobutanol production yield by threefold. In aerobic cultivations in shake flasks in mineral medium, the isobutanol yield of the ILV2 ILV3 ILV5 overexpression strain and the reference strain were 3.86 and 0.28 mg per g glucose, respectively. They increased to 4.12 and 2.4 mg per g glucose in yeast extract/peptone/dextrose (YPD complex medium under aerobic conditions, respectively. Conclusions Overexpression of genes ILV2, ILV3, ILV5, and BAT2 in valine metabolism led to an increase in isobutanol production in S. cerevisiae. Additional overexpression of ILV6 in the ILV2 ILV3 ILV5 overexpression strain had a negative effect, presumably by increasing the sensitivity of Ilv2 to valine inhibition, thus weakening the positive impact of overexpression of ILV2, ILV3, and ILV5 on isobutanol production. Aerobic cultivations of the ILV2 ILV3 ILV5 overexpression strain and the reference strain showed that supplying amino acids in cultivation media gave a substantial improvement in isobutanol production for the reference strain, but not for the ILV2 ILV3 ILV5 overexpression strain. This result implies that other constraints besides the enzyme activities for the supply of 2-ketoisovalerate may become bottlenecks for isobutanol production after ILV2, ILV3, and ILV5 have been overexpressed, which most probably includes the valine inhibition to Ilv2.

  4. Increased isobutanol production in Saccharomyces cerevisiae by overexpression of genes in valine metabolism

    DEFF Research Database (Denmark)

    Chen, Xiao; Nielsen, Kristian Fog

    2011-01-01

    BACKGROUND: Isobutanol can be a better biofuel than ethanol due to its higher energy density and lower hygroscopicity. Furthermore, the branched-chain structure of isobutanol gives a higher octane number than the isomeric n-butanol. Saccharomyces cerevisiae was chosen as the production host because of its relative tolerance to alcohols, robustness in industrial fermentations, and the possibility for future combination of isobutanol production with fermentation of lignocellulosic materials. RESULTS: The yield of isobutanol was improved from 0.16 to 0.97 mg per g glucose by simultaneous overexpression of biosynthetic genes ILV2, ILV3, and ILV5 in valine metabolism in anaerobic fermentation of glucose in mineral medium in S. cerevisiae. Isobutanol yield was further improved by twofold by the additional overexpression of BAT2, encoding the cytoplasmic branched-chain amino-acid aminotransferase. Overexpression of ILV6, encoding the regulatory subunit of Ilv2, in the ILV2 ILV3 ILV5 overexpression strain decreased isobutanol production yield by threefold. In aerobic cultivations in shake flasks in mineral medium, the isobutanol yield of the ILV2 ILV3 ILV5 overexpression strain and the reference strain were 3.86 and 0.28 mg per g glucose, respectively. They increased to 4.12 and 2.4 mg per g glucose in yeast extract/peptone/dextrose (YPD) complex medium under aerobic conditions, respectively. CONCLUSIONS: Overexpression of genes ILV2, ILV3, ILV5, and BAT2 in valine metabolism led to an increase in isobutanol production in S. cerevisiae. Additional overexpression of ILV6 in the ILV2 ILV3 ILV5 overexpression strain had a negative effect, presumably by increasing the sensitivity of Ilv2 to valine inhibition, thus weakening the positive impact of overexpression of ILV2, ILV3, and ILV5 on isobutanol production. Aerobic cultivations of the ILV2 ILV3 ILV5 overexpression strain and the reference strain showed that supplying amino acids in cultivation media gave a substantial improvement in isobutanol production for the referencestrain, but not for the ILV2 ILV3 ILV5 overexpression strain. This result implies that other constraints besides the enzyme activities for the supply of 2-ketoisovalerate may become bottlenecks for isobutanol production after ILV2, ILV3, and ILV5 have been overexpressed, which most probably includes the valine inhibition to Ilv2.

  5. Expresión heteróloga de un péptido multiepitópico de células B de M. tuberculosis en Saccharomyces cerevisiae

    Scientific Electronic Library Online (English)

    María de los Angeles, García; María Elena, Sarmiento; Roberto, Coria; Laura, Kawasaky; Laura, Ongay; Juan Francisco, de la Rosa; Armando, Acosta.

    2007-08-01

    Full Text Available Saccharomyces cerevisiae ha sido ampliamente utilizada como sistema de expresión de proteínas heterólogas. El presente trabajo se encaminó hacia la expresión en Saccharomyces cerevisiae de un péptido de epitopes múltiples de M. tuberculosis. Con dicho propósito el péptido quimérico denominado B2 fue [...] clonado en dos vectores de expresión de esta levadura con promotores regulables por galactosa y sulfato cúprico, respectivamente. Luego de los experimentos de inducción, la expresión del péptido B2 fue analizada mediante SDS/PAGE y Western blot. El análisis por Western blot confirmó la expresión del péptido B2, al hacerse la inducción con 100 mM de CuSO4 durante toda la noche. No ocurrió así en los experimentos donde se utilizó la galactosa como inductor con todas las condiciones ensayadas. Estos resultados muestran que la levadura Saccharomyces cerevisiae pudiera ser un buen hospedero alternativo para la expresión de péptidos multiepitópicos de M. tuberculosis. Abstract in english Saccharomyces cerevisiae has been widely used as expression system of heterologous proteins. The aim of the present work was the expression in Saccharomyces cerevisiae of a class B multiepitopic peptide from M. tuberculosis. For this purpose, the chimerical peptide named B2 was cloned in two yeast e [...] xpression vectors containing galactose and cupric sulphate regulated promoters, respectively. After induction experiments, B2 expression was analyzed by SDS/PAGE and Western Blot. By Western Blot analysis B2 expression was confirmed when induction took place overnight with 100 mM of CuSO4. No expression signal took place when galactose was used as inductor. These results show that Sacchromyces cervisiae could be a good alternative host for the expression of multiepitopic peptides from M. tuberculosis.

  6. Radiosensitivity of Saccharomyces cerevisiae W303-1A and BY4741 Strains

    International Nuclear Information System (INIS)

    Saccharomyces cerevisiae, a simple eukaryotic cell, has been widely used as a model for all eukaryotes including humans for the study of fundamental cellular processes such as DNA replication, DNA recombination, cell cycle, cell division and metabolism. Numerous laboratory strains are used in yeast research. Most of the mutants have been derived from the two widely used laboratory strains W303-1A and BY4741. While BY4741 is a derivative of S288C, used in the systematic sequencing of the S. cerevisiae genome, strains with a W303 background serve in many physiological and biochemical studies. It was found in a recent study that W303-1A contains a mutant allele of YBP1, ybp1-1, encoding four amino acid substitutions, that results in increased peroxide sensitivity. Mutation of ybp1-1 is not a complete loss of function allele as it is more resistant to peroxides than the knock-out mutant. Ybp1 is required for oxidation of specific cysteine residues of the transcription factor Yap1p resulting in the nuclear localization of Yap1p in response to stress. Ionizing radiation (IR) can produce highly reactive hydroxyl radicals through the decomposition of cellular water, such as superoxide anion radical, hydrogen peroxide, hydroxyl radical. These reactive oxygen species (ROS) can cause wide-ranging cellular damage, including DNA double-strand breaks (DSBs), lipid peroxidation, and protein modification. Also, ROS produced by IR cause oxidative stress. Detoxification enzymes are active stress. Detoxification enzymes are activated for ROS scavenging against oxidative stress. Also, antioxidants are used for detoxification of ROS and reduction of oxidative damage. NAC, one of the antioxidants, is a precursor for glutathione (GSH). The aim of the present study was to compare the differences in radiosensitivity associated cell viability between the two strains. Also, effect of NAC against IR on cell protection was investigated

  7. Radiosensitivity of Saccharomyces cerevisiae W303-1A and BY4741 Strains

    Energy Technology Data Exchange (ETDEWEB)

    Park, Ji Young; Kim, Jin Kyu [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of); Nili, Mohammad [Dawnesh Radiation Research Institute, Barcelona (Spain)

    2011-05-15

    Saccharomyces cerevisiae, a simple eukaryotic cell, has been widely used as a model for all eukaryotes including humans for the study of fundamental cellular processes such as DNA replication, DNA recombination, cell cycle, cell division and metabolism. Numerous laboratory strains are used in yeast research. Most of the mutants have been derived from the two widely used laboratory strains W303-1A and BY4741. While BY4741 is a derivative of S288C, used in the systematic sequencing of the S. cerevisiae genome, strains with a W303 background serve in many physiological and biochemical studies. It was found in a recent study that W303-1A contains a mutant allele of YBP1, ybp1-1, encoding four amino acid substitutions, that results in increased peroxide sensitivity. Mutation of ybp1-1 is not a complete loss of function allele as it is more resistant to peroxides than the knock-out mutant. Ybp1 is required for oxidation of specific cysteine residues of the transcription factor Yap1p resulting in the nuclear localization of Yap1p in response to stress. Ionizing radiation (IR) can produce highly reactive hydroxyl radicals through the decomposition of cellular water, such as superoxide anion radical, hydrogen peroxide, hydroxyl radical. These reactive oxygen species (ROS) can cause wide-ranging cellular damage, including DNA double-strand breaks (DSBs), lipid peroxidation, and protein modification. Also, ROS produced by IR cause oxidative stress. Detoxification enzymes are activated for ROS scavenging against oxidative stress. Also, antioxidants are used for detoxification of ROS and reduction of oxidative damage. NAC, one of the antioxidants, is a precursor for glutathione (GSH). The aim of the present study was to compare the differences in radiosensitivity associated cell viability between the two strains. Also, effect of NAC against IR on cell protection was investigated

  8. Isolation and characterization of saccharomyces cerevisiae RFA1 gene mutations which confer ultraviolet light sensitivity

    International Nuclear Information System (INIS)

    Replication protein A (RPA) has been identified as an essential factor of SV40 DNA replication in vitro and is involved in DNA replication, recombination, nucleotide excision repair (NER), mismatch repair, and checkpoint control. It is a heterotrimeric single-stranded DNA binding protein consisting of 70-, 34-, and 14-kDa subunits and highly conserved among eukaryotes, In Saccharomyces cerevisiae, ultraviolet light (UV) sensitive mutations on RFA1 gene which encodes the largest subunit of RPA have been isolated. UV sensitivity could be caused by the defect of several mechanisms, including NER, recombinational repair, and checkpoint control. However, it is unclear which mechanism is defective by each mutation. To investigate it, mutations which confer UV sensitivity on S. cerevisiae RFA1 gene were searched using plasmid shuffling method. Two mutations (36+60 and t6) were isolated and m51 mutation, previously reported by Umezu et al., was added to further analysis. The strain carrying each rfa1 allele showed not only UV sensitivity but also methyl methane sulfonate sensitivity. By the epistasis with rad14, rad52, and rad6 mutations, all mutations were epistatic to RAD52, not to RAD14 or RAD6. UV sensitivity of each rfa1 strain was partially suppressed by over-expression of RAD52 gene. Partial suppression for UV sensitivity by over-expression of each rfa1 gene was also seen in m51 and t6 strains. The amount of RPA was predominantly reduced in m51 and t6 strains. Taken toly reduced in m51 and t6 strains. Taken together, these rfa1 gene mutations cause the defect of recombinational repair. The defect in m51 and t6 strains is mainly caused by the reduction of RPA, while the defect in 36+60 strain is caused by other reasons. (author)

  9. Constitutive optimized production of streptokinase in Saccharomyces cerevisiae utilizing glyceraldehyde 3-phosphate dehydrogenase promoter of Pichia pastoris.

    Science.gov (United States)

    Vellanki, Ravi N; Potumarthi, Ravichandra; Doddapaneni, Kiran K; Anubrolu, Naveen; Mangamoori, Lakshmi N

    2013-01-01

    A novel expression vector constructed from genes of Pichia pastoris was applied for heterologous gene expression in Saccharomyces cerevisiae. Recombinant streptokinase (SK) was synthesized by cloning the region encoding mature SK under the control of glyceraldehyde 3-phosphate dehydrogenase (GAP) promoter of Pichia pastoris in Saccharomyces cerevisiae. SK was intracellularly expressed constitutively, as evidenced by lyticase-nitroanilide and caseinolytic assays. The functional activity was confirmed by plasminogen activation assay and in vitro clot lysis assay. Stability and absence of toxicity to the host with the recombinant expression vector as evidenced by southern analysis and growth profile indicate the application of this expression system for large-scale production of SK. Two-stage statistical approach, Plackett-Burman (PB) design and response surface methodology (RSM) was used for SK production medium optimization. In the first stage, carbon and organic nitrogen sources were qualitatively screened by PB design and in the second stage there was quantitative optimization of four process variables, yeast extract, dextrose, pH, and temperature, by RSM. PB design resulted in dextrose and peptone as best carbon and nitrogen sources for SK production. RSM method, proved as an efficient technique for optimizing process conditions which resulted in 110% increase in SK production, 2352 IU/mL, than for unoptimized conditions. PMID:24171161

  10. Transcription factor Stb5p is essential for acetaldehyde tolerance in Saccharomyces cerevisiae.

    Science.gov (United States)

    Matsufuji, Yoshimi; Nakagawa, Tomoyuki; Fujimura, Shuki; Tani, Akio; Nakagawa, Junichi

    2010-10-01

    Transcription factor Stb5p, previously known as one of the multidrug resistance gene regulators in Saccharomyces cerevisiae, was shown here to play an essential role in acetaldehyde tolerance. A mutant strain, ?stb5 exhibited increased acetaldehyde sensitivity, and failed to induce genes such as GND1, TKL1 and TAL1 involved in the pentose phosphate pathway (PPP) upon acetaldehyde stress. Using this strain it was revealed that Stb5p acts as a repressor for PGI1 encoding glucose-6-phosphate isomerase under acetaldehyde stress. In reverse, over-expression of Stb5p reinforced acetaldehyde tolerance to the yeast. Furthermore, various deletion mutants of the genes involved in glycolysis showed increased acetaldehyde tolerance compared to the wild-type strain. From these results, it was suggested that Stb5p participates in acetaldehyde tolerance by regulating expression of the PPP genes and PGI1, and that down-regulation of glycolytic pathway may lead to vitalization of PPP and to increased acetaldehyde tolerance. PMID:20806246

  11. Gene dosage alteration of L2 ribosomal protein genes in Saccharomyces cerevisiae: effects on ribosome synthesis.

    Science.gov (United States)

    Lucioli, A; Presutti, C; Ciafrè, S; Caffarelli, E; Fragapane, P; Bozzoni, I

    1988-01-01

    In Saccharomyces cerevisiae, the genes coding for the ribosomal protein L2 are present in two copies per haploid genome. The two copies, which encode proteins differing in only a few amino acids, contribute unequally to the L2 mRNA pool: the L2A copy makes 72% of the mRNA, while the L2B copy makes only 28%. Disruption of the L2B gene (delta B strain) did not lead to any phenotypic alteration, whereas the inactivation of the L2A copy (delta A strain) produced a slow-growth phenotype associated with decreased accumulation of 60S subunits and ribosomes. No intergenic compensation occurred at the transcriptional level in the disrupted strains; in fact, delta A strains contained reduced levels of L2 mRNA, whereas delta B strains had almost normal levels. The wild-type phenotype was restored in the delta A strains by transformation with extra copies of the intact L2A or L2B gene. As already shown for other duplicated genes (Kim and Warner, J. Mol. Biol. 165:79-89, 1983; Leeret al., Curr. Genet. 9:273-277, 1985), the difference in expression of the two gene copies could be accounted for via differential transcription activity. Sequence comparison of the rpL2 promoter regions has shown the presence of canonical HOMOL1 boxes which are slightly different in the two genes. Images PMID:3062369

  12. ATP16 Genes and Neighboring ORFs Are Duplicated on Chromosome IV in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Masaharu Takeda

    2011-12-01

    Full Text Available We present evidence that there were two closely linked copies of the ATP16 (YDL004w gene encoding the delta subunit of F1F0-ATPase complex on chromosome IV in laboratory strains, W303-1A, W303-1B, DC5, LL20, SEY2102, YPH499, and S288C of Saccharomyces cerevisiae. We previously reported that there were 2-3 copies of ATP1 (alpha, ATP2 (beta and ATP3 (gamma on chromosomes II, X, and II, respectively. Homologous recombination of the ATP16 with HIS3 (YOR202w, gene walking, and long-PCR analyses showed that two ATP16 were present on the same chromosome as above genes. The gene walking estimated that the two ATP16 were separated by approximately 8.4 kb using ATP16 and its neighboring DNAs as probes, designated the proximal ATP16 to the telomere as ATP16a and to the distal as ATP16b. Although the nucleotide sequences of ATP16a and ATP16b were identical including 833 bases upstream- and 937 bases downstream of ATP16, they might be expressed differently.

  13. DAL82, a second gene required for induction of allantoin system gene transcription in Saccharomyces cerevisiae.

    Science.gov (United States)

    Olive, M G; Daugherty, J R; Cooper, T G

    1991-01-01

    Several highly inducible enzyme activities are required for the degradation of allantoin in Saccharomyces cerevisiae. Induction of these pathway enzymes has been shown to be regulated at transcription, and response to inducer is lost in dal81 and dal82/durM mutants. The similar phenotypes generated by dal81 and dal82 mutations prompted the question of whether they were allelic. We demonstrated that the DAL81 and DAL82 loci are distinct, unlinked genes situated on chromosomes IX and XIV. DAL82 gene expression did not respond to induction by the allantoin pathway inducer or to nitrogen catabolite repression. Expression was also not significantly affected by mutation of the dal80 locus. From the nucleotide sequence of the DAL82 gene, we deduced that it encodes a protein with a mass of 29,079 Da that may possess the structural motifs expected of a regulatory protein. This protein was shown to be required for the function mediated by the cis-acting upstream induction sequence situated in the 5'-flanking regions of the inducible allantoin pathway genes. Images PMID:1898922

  14. Optimization of ordered plasmid assembly by gap repair in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Eckert-Boulet, Nadine Valerie; Pedersen, Mette Louise

    2012-01-01

    Combinatorial genetic libraries are powerful tools for diversifying and optimizing biomolecules. The process of library assembly is a major limiting factor for library complexity and quality. Gap repair by homologous recombination in Saccharomyces cerevisiae can facilitate in vivo assembly of DNA fragments sharing short patches of sequence homology, thereby supporting generation of high-complexity libraries without compromising fidelity. In this study, we have optimized the ordered assembly of three DNA fragments into a gapped vector by in vivo homologous recombination. Assembly is achieved by co-transformation of the DNA fragments and the gapped vector, using a modified lithium acetate protocol. The optimal gap-repair efficiency is found at a 1:80 molar ratio of gapped vector to each of the three fragments. We measured gap-repair efficiency in different genetic backgrounds and observed increased efficiency in mutants carrying a deletion of the SGS1 helicase-encoding gene. Using our experimental conditions, agap-repair efficiency of > 10(6) plasmid-harbouring colonies/µg gapped vector DNA is obtained in a single transformation, with a recombination fidelity > 90%.

  15. ATPase and DNA helicase activities of the Saccharomyces cerevisiae anti-recombinase Srs2.

    Science.gov (United States)

    Van Komen, Stephen; Reddy, Mothe Sreedhar; Krejci, Lumir; Klein, Hannah; Sung, Patrick

    2003-11-01

    Saccharomyces cerevisiae SRS2 encodes an ATP-dependent DNA helicase that is needed for DNA damage checkpoint responses and that modulates the efficiency of homologous recombination. Interestingly, strains simultaneously mutated for SRS2 and a variety of DNA repair genes show low viability that can be overcome by inactivating homologous recombination, thus implicating inappropriate recombination as the cause of growth impairment in these mutants. Here, we report on our biochemical characterization of the ATPase and DNA helicase activities of Srs2. ATP hydrolysis by Srs2 occurs efficiently only in the presence of DNA, with ssDNA being considerably more effective than dsDNA in this regard. Using homopolymeric substrates, the minimal DNA length for activating ATP hydrolysis is found to be 5 nucleotides, but a length of 10 nucleotides is needed for maximal activation. In its helicase action, Srs2 prefers substrates with a 3' ss overhang, and approximately 10 bases of 3' overhanging DNA is needed for efficient targeting of Srs2 to the substrate. Even though a 3' overhang serves to target Srs2, under optimized conditions blunt-end DNA substrates are also dissociated by this protein. The ability of Srs2 to unwind helicase substrates with a long duplex region is enhanced by the inclusion of the single-strand DNA-binding factor replication protein A. PMID:12966095

  16. DNA binding properties of the Saccharomyces cerevisiae DAT1 gene product.

    Science.gov (United States)

    Reardon, B J; Gordon, D; Ballard, M J; Winter, E

    1995-12-11

    The DAT1 gene of Saccharomyces cerevisiae encodes a DNA binding protein (Dat1p) that specifically recognizes the minor groove of non-alternating oligo(A).oligo(T) tracts. Sequence-specific recognition requires arginine residues found within three perfectly repeated pentads (G-R-K-P-G) of the Dat1p DNA binding domain [Reardon, B. J., Winters, R. S., Gordon, D., and Winter, E. (1993) Proc. Natl. Acad. Sci. USA 90, 11327-1131]. This report describes a rapid and simple method for purifying the Dat1p DNA binding domain and the biochemical characterization of its interaction with oligo(A).oligo(T) tracts. Oligonucleotide binding experiments and the characterization of yeast genomic Dat1p binding sites show that Dat1p specifically binds to any 11 base sequence in which 10 bases conform to an oligo(A).oligo(T) tract. Binding studies of different sized Dat1p derivatives show that the Dat1p DNA binding domain can function as a monomer. Competition DNA binding assays using poly(I).poly(C), demonstrate that the minor groove oligo(A).oligo(T) constituents are not sufficient for high specificity DNA binding. These data constrain the possible models for Dat1p/oligo(A).oligo(T) complexes, suggest that the DNA binding domain is in an extended structure when complexed to its cognate DNA, and show that Dat1p binding sites are more prevalent than previously thought. PMID:8532535

  17. Quantitative modeling of the Saccharomyces cerevisiae FLR1 regulatory network using an S-system formalism.

    Science.gov (United States)

    Calçada, Dulce; Vinga, Susana; Freitas, Ana T; Oliveira, Arlindo L

    2011-10-01

    In this study we address the problem of finding a quantitative mathematical model for the genetic network regulating the stress response of the yeast Saccharomyces cerevisiae to the agricultural fungicide mancozeb. An S-system formalism was used to model the interactions of a five-gene network encoding four transcription factors (Yap1, Yrr1, Rpn4 and Pdr3) regulating the transcriptional activation of the FLR1 gene. Parameter estimation was accomplished by decoupling the resulting system of nonlinear ordinary differential equations into a larger nonlinear algebraic system, and using the Levenberg-Marquardt algorithm to fit the models predictions to experimental data. The introduction of constraints in the model, related to the putative topology of the network, was explored. The results show that forcing the network connectivity to adhere to this topology did not lead to better results than the ones obtained using an unrestricted network topology. Overall, the modeling approach obtained partial success when trained on the nonmutant datasets, although further work is required if one wishes to obtain more accurate prediction of the time courses. PMID:21976379

  18. A synthetic suicide riboswitch for the high-throughput screening of metabolite production in Saccharomyces cerevisiae.

    Science.gov (United States)

    Lee, Sang-Woo; Oh, Min-Kyu

    2015-03-01

    Artificial devices such as the synthetic riboswitch have shown potential to introduce unnatural phenotypic perturbation because its synthetic traits are distinct from that of innate metabolism. In this study, a riboswitch, a small regulatory element found in RNAs, was employed to reprogram microorganisms to produce valuable metabolites. A self-cleaving ribozyme glmS, found in gram-positive bacteria, cleaves its own transcript in response to the intracellular glucosamine 6-phosphate (GlcN6P) concentration. The glmS ribozyme was integrated into the 3'-untranslated region of FCY1, which encodes cytosine deaminase in Saccharomyces cerevisiae to construct a suicide riboswitch for evolutionary engineering. Growth of the strain harboring the suicide riboswitch was hampered by the addition of fluorocytosine, and was recovered as metabolite level increased. By using this riboswitch, we isolated a N-acetyl glucosamine (GlcNAc) producer strain by screening an efficient glutamine-fructose-6-phosphate transaminase (Gfa1p) and haloacid dehalogenase-like phosphatases (HAD phosphatases) originated from Escherichia coli. The suicide riboswitch was also applied to different metabolite by using artificial allosteric ribozyme. Since the mechanisms used in this work are universal in microorganisms, our synthetic suicide riboswitch can be applied to a wide range of organisms and can be exploited to the efficient and high-throughput screening of inconspicuous phenotypes. PMID:25596509

  19. Gateway vectors for efficient artificial gene assembly in vitro and expression in yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Giuraniuc, Claudiu V; MacPherson, Murray; Saka, Yasushi

    2013-01-01

    Construction of synthetic genetic networks requires the assembly of DNA fragments encoding functional biological parts in a defined order. Yet this may become a time-consuming procedure. To address this technical bottleneck, we have created a series of Gateway shuttle vectors and an integration vector, which facilitate the assembly of artificial genes and their expression in the budding yeast Saccharomyces cerevisiae. Our method enables the rapid construction of an artificial gene from a promoter and an open reading frame (ORF) cassette by one-step recombination reaction in vitro. Furthermore, the plasmid thus created can readily be introduced into yeast cells to test the assembled gene's functionality. As flexible regulatory components of a synthetic genetic network, we also created new versions of the tetracycline-regulated transactivators tTA and rtTA by fusing them to the auxin-inducible degron (AID). Using our gene assembly approach, we made yeast expression vectors of these engineered transactivators, AIDtTA and AIDrtTA and then tested their functions in yeast. We showed that these factors can be regulated by doxycycline and degraded rapidly after addition of auxin to the medium. Taken together, the method for combinatorial gene assembly described here is versatile and would be a valuable tool for yeast synthetic biology. PMID:23675537

  20. Alleviating Redox Imbalance Enhances 7-Dehydrocholesterol Production in Engineered Saccharomyces cerevisiae

    Science.gov (United States)

    Liu, Duo; Zhou, Xiao; Yuan, Ying-Jin

    2015-01-01

    Maintaining redox balance is critical for the production of heterologous secondary metabolites, whereas on various occasions the native cofactor balance does not match the needs in engineered microorganisms. In this study, 7-dehydrocholesterol (7-DHC, a crucial precursor of vitamin D3) biosynthesis pathway was constructed in Saccharomyces cerevisiae BY4742 with endogenous ergosterol synthesis pathway blocked by knocking out the erg5 gene (encoding C-22 desaturase). The deletion of erg5 led to redox imbalance with higher ratio of cytosolic free NADH/NAD+ and more glycerol and ethanol accumulation. To alleviate the redox imbalance, a water-forming NADH oxidase (NOX) and an alternative oxidase (AOX1) were employed in our system based on cofactor regeneration strategy. Consequently, the production of 7-dehydrocholesterol was increased by 74.4% in shake flask culture. In the meanwhile, the ratio of free NADH/NAD+ and the concentration of glycerol and ethanol were reduced by 78.0%, 50.7% and 7.9% respectively. In a 5-L bioreactor, the optimal production of 7-DHC reached 44.49(±9.63) mg/L. This study provides a reference to increase the production of some desired compounds that are restricted by redox imbalance. PMID:26098102

  1. Homologous Recombination Repair Within the rDNA Array in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    M. Motovali-Bashi

    2007-01-01

    Full Text Available Homologous recombination repair starts with Double-strand Breaks (DSBs followed by crossing-over and recombination. The expected frequency of meiotic chromosomal exchange in the region of chromosome XII encoding ribosomal DNA in Saccharomyces cerevisiae is 3.5 to 5 events per cell per meiosis. However interchromosomal meiotic recombination in the rDNA gene is very rare, suggesting repression of DSB and crossing-over. On the other hand, mitotic events such as intrachromosomal recombination producing 3 ?m rDNA circles (which accumulate with cellular age and unequal sister chromatid exchanges appear to be quite common. This study looked at the rDNA breakage in the strain ORD 1181, a rad50S mutant with SK1 background, which does a relatively fast and near synchronous meiosis. The fine analysis of the rDNA array was performed using restriction endonuclease enzymes that do not cleave within the rDNA array. The results suggest that there are at least two hot regions for chromosome breakage within the rDNA array. According to our previous studies we suggest that the DSB hot regions are in one homologue. However, there is possibility that other homologue is involving in DSB too.

  2. Multidrug resistance transporters Snq2p and Pdr5p mediate caffeine efflux in Saccharomyces cerevisiae.

    Science.gov (United States)

    Tsujimoto, Yoshiyuki; Shimizu, Yoshihiro; Otake, Kazuya; Nakamura, Tatsuya; Okada, Ryutaro; Miyazaki, Toshitaka; Watanabe, Kunihiko

    2015-07-01

    SNQ2 was identified as a caffeine-resistance gene by screening a genomic library of Saccharomyces cerevisiae in a multicopy vector YEp24. SNQ2 encodes an ATP-binding cassette transporter and is highly homologous to PDR5. Multicopy of PDR5 also conferred resistance to caffeine, while its resistance was smaller than that of SNQ2. Residual caffeine contents were analyzed after transiently exposing cells to caffeine. The ratios of caffeine contents were 21.3?±?8.8% (YEp24-SNQ2) and 81.9?±?8.7% (YEp24-PDR5) relative to control (YEp24, 100%). In addition, multicopies of SNQ2 or PDR5 conferred resistance to rhodamine 6G (R6G), which was widely used as a substrate for transport assay. R6G was exported by both transporters, and their efflux activities were inhibited by caffeine with half-maximal inhibitory concentrations of 5.3 ± 1.9 (YEp24-SNQ2) and 17.2 ± 9.6 mM (YEp24-PDR5). These results demonstrate that Snq2p is a more functional transporter of caffeine than Pdr5p in yeast cells. PMID:25686090

  3. Nucleoside diphosphate kinase of Saccharomyces cerevisiae, Ynk1p: localization to the mitochondrial intermembrane space.

    Science.gov (United States)

    Amutha, Boominathan; Pain, Debkumar

    2003-03-15

    Nucleoside diphosphate kinase (NDPK) is a highly conserved multifunctional enzyme. It catalyses the transfer of gamma phosphates from nucleoside triphosphates to nucleoside diphosphates by a mechanism that involves formation of an autophosphorylated enzyme intermediate. The phosphate is usually supplied by ATP. NDPK activity in different subcellular compartments may regulate the crucial balance between ATP and GTP or other nucleoside triphosphates. NDPKs are homo-oligomeric proteins and are predominantly localized in the cytosol. In this paper, we demonstrate that in Saccharomyces cerevisiae a small fraction of total NDPK activity encoded by YNK1 is present in the intermembrane space (IMS) of mitochondria, and the corresponding protein Ynk1p in the IMS represents approx. 0.005% of total mitochondrial proteins. Ynk1p, synthesized as a single gene product, must therefore be partitioned between cytoplasm and mitochondrial IMS fractions. A mechanism for this partitioning is suggested by our observations that interaction with a 40 kDa protein of the translocase of outer mitochondrial membrane (Tom40p), occurs preferentially with unfolded, unphosphorylated forms of Ynk1p. A population of newly translated, but not yet folded or autophosphorylated, Ynk1p intermediates may be imported into the IMS of mitochondria and trapped there by subsequent folding and oligomerization. Within the small volume of the IMS, Ynk1p may be more concentrated and may be required to supply GTP to several important proteins in this compartment. PMID:12472466

  4. Automated Yeast Mating Protocol Using Open Reading Frames from Saccharomyces cerevisiae Genome to Improve Yeast Strains for Cellulosic Ethanol Production

    Science.gov (United States)

    Engineering the industrial ethanologen Saccharomyces cerevisiae to utilize pentose sugars from lignocellulosic biomass is critical for commercializing cellulosic fuel ethanol production. Approaches to engineer pentose-fermenting yeasts have required expression of additional genes. We implemented a...

  5. Systems Biology of Saccharomyces cerevisiae Physiology and its DNA Damage Response

    DEFF Research Database (Denmark)

    Fazio, Alessandro

    2010-01-01

    The yeast Saccharomyces cerevisiae is a model organism in biology, being widely used in fundamental research, the first eukaryotic organism to be fully sequenced and the platform for the development of many genomics techniques. Therefore, it is not surprising that S. cerevisiae has also been widely used in the field of systems biology during the last decade. This thesis investigates S. cerevisiae growth physiology and DNA damage response by using a systems biology approach. Elucidation of the relationship between growth rate and gene expression is important to understand the mechanisms regulating cell growth. In order to study this relationship, we have grown S. cerevisiae cells in chemostat at defined growth rates and measured the transcriptional response. We have applied a complex experimental design, involving three factors: specific growth rate, oxygen availability and nutrient limitation. We have identified 268 growth rate dependent genes. These genes were used to identify key areas of the metabolism around which expression changes were significantly associated and we found nucleotide synthesis and ATP producing and consuming reactions. Moreover, by scoring the significance of overlap between growth rate dependent genes and known transcription factor (TF) target sets, we identified 13 TFs, involved in stress response, cell cycle and ribosome biogenesis, that appeared to coordinate the response at increasing growth rates. Therefore, in this study we have identified a more conservative set of growth dependent genes by using a multi-factorial experimental design. Moreover, new insights into the metabolic response and transcriptional regulation of these genes have been provided by using systems biology tools (Chapter 3). One of the prerequisite of systems biology should be the standardization and reproducibility of experimental and analytical techniques, in order to allow the comparison of data generated in different laboratories. With the aim of addressing this aspect, we have collaborated in a large study involving ten laboratories, constituting the Yeast Systems Biology Network (YSBN). S. cerevisiae cultivations were performed in a single laboratory and samples were sent to the other partners. The experimental design involved two factors: strain (CEN.PK113-7D and YSBN2) and growth condition (batch and chemostat). Transcriptome was measured with four different platforms (Affymetrix, Agilent, qPCR and TRAC), metabolome was analyzed in seven laboratories, using different protocols, and enzyme activities were determined in two different laboratories. The comparison of the analyses showed that reproducibility of the results was affected by the laboratory and the protocol used. Transcription and enzyme activity analyses gave consistent results, while metabolite level measurements showed some variability. Therefore, even though the source of biomass was unique, the reproducibility of data appeared to be a challenging task. Nevertheless, we were able to perform an integrative analysis and discover that the lower biomass yield of CEN.PK113-7D was due to higher protein turnover than YSBN2; this finding would not be achievable using a single omics dataset. Moreover, the generated datasets are a valuable resource for the yeast systems biology community (Chapter 4). Upon DNA damage, S. cerevisiae cells respond activating the so-called cell cycle checkpoints that promote damage repair and viability. The activation of these checkpoints depends on kinase cascades and regulation of transcription is one of the responses elicited by checkpoint activation. Therefore, we have decided to investigate the transcriptional and phenotypic responses to the alkylating agent methyl methanesulfonate (MMS) of mutant strains carrying deletions of genes encoding protein kinases (Mec1, Tel1, Rad53, Dun1, Chk1, Alk1) and protein phosphatases (Ptc3, Pph3, Oca1) involved in DNA damage response (DDR). We have discovered a prominent role for Rad53, Mec1 and Tel1 in transcriptional response. Moreover, we have shown for the first time the important role of O

  6. Microbial cells as biosorbents for heavy metals: accumulation of Uranium by Saccharomyces cerevisiae and Pseudomonas aeruginosa

    International Nuclear Information System (INIS)

    Uranium accumulated extracellularly on the surfaces of Saccharomyces cerevisiae cells. The rate and extent of accumulation were subject to environmental parameters, such as pH, temperature, and interference by certain anions and cations. Uranium accumulation by Pseudomonas aeruginosa occurred intracellularly and was extremely rapid (<10 s), and no response to environmental parameters could be detected. Metabolism was not required for metal uptake by either organism. Cell-bound uranium reached a concentration of 10 to 15% of the dry cell weight, but only 32% of the S. cerevisiae cells and 44% of the P. aeruginosa cells within a given population possessed visible uranium deposits when examined by electron microscopy. Rates of uranium uptake by S. cerevisiae were increased by chemical pretreatment of the cells. Uranium could be removed chemically from S. cerevisiae cells, and the cells could then be reused as a biosorbent

  7. Ecological success of a group of Saccharomyces cerevisiae/Saccharomyces kudriavzevii hybrids in the northern european wine-making environment.

    Science.gov (United States)

    Erny, C; Raoult, P; Alais, A; Butterlin, G; Delobel, P; Matei-Radoi, F; Casaregola, S; Legras, J L

    2012-05-01

    The hybrid nature of lager-brewing yeast strains has been known for 25 years; however, yeast hybrids have only recently been described in cider and wine fermentations. In this study, we characterized the hybrid genomes and the relatedness of the Eg8 industrial yeast strain and of 24 Saccharomyces cerevisiae/Saccharomyces kudriavzevii hybrid yeast strains used for wine making in France (Alsace), Germany, Hungary, and the United States. An array-based comparative genome hybridization (aCGH) profile of the Eg8 genome revealed a typical chimeric profile. Measurement of hybrids DNA content per cell by flow cytometry revealed multiple ploidy levels (2n, 3n, or 4n), and restriction fragment length polymorphism analysis of 22 genes indicated variable amounts of S. kudriavzevii genetic content in three representative strains. We developed microsatellite markers for S. kudriavzevii and used them to analyze the diversity of a population isolated from oaks in Ardèche (France). This analysis revealed new insights into the diversity of this species. We then analyzed the diversity of the wine hybrids for 12 S. cerevisiae and 7 S. kudriavzevii microsatellite loci and found that these strains are the products of multiple hybridization events between several S. cerevisiae wine yeast isolates and various S. kudriavzevii strains. The Eg8 lineage appeared remarkable, since it harbors strains found over a wide geographic area, and the interstrain divergence measured with a (??)(2) genetic distance indicates an ancient origin. These findings reflect the specific adaptations made by S. cerevisiae/S. kudriavzevii cryophilic hybrids to winery environments in cool climates. PMID:22344648

  8. Biotransformation of Carmoisine and Reactive Black 5 Dyes Using Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Abbas Sadeghi

    2014-04-01

    Full Text Available Saccharomyces cerevisiae (baker’s yeast is the most important industrial microorganisms. This yeast is commonly used as a leavening agent in baking bread and bakery products, where it produces carbon dioxide from converting of the fermentable sugars present in the dough. Nowadays, industrial and chemical activities led to produce new compounds with new kinds of contamination in the environment. Discharge of untreated or partially treated industrial sewage has created the contamination problems of rivers and lakes such as drugs, oil, heavy metals, paints, pesticides and various chemical compounds in them. Hence, it is necessary to control and reduce the levels of these compounds in wastewater and bring them to permissible values. This study aims to study the bioconversion potential of commonly available Saccharomyces cerevisiae for the two textile dyes of Carmoisine and Reactive Black 5. Reaction mixtures for biotransformation of dyes included 50 mg/l Carmoisine or 25 mg/l Reactive Black 5 and 1% dried harvested cells of S. cerevisiae (bread’s yeast were tested. Harvested dry and wet yeast were studied for this purpose. The results show that harvested cells of Saccharomyces cerevisiae are able to bioconvert Carmoisine and Reactive Black 5. Reactive Black 5, Carmoisine are degraded by biotransformation 85% and 53% within 24 hours in water at the room temperature.

  9. Performance evaluation of Pichia kluyveri, Kluyveromyces marxianus and Saccharomyces cerevisiae in industrial tequila fermentation.

    Science.gov (United States)

    Amaya-Delgado, L; Herrera-López, E J; Arrizon, Javier; Arellano-Plaza, M; Gschaedler, A

    2013-05-01

    Traditionally, industrial tequila production has used spontaneous fermentation or Saccharomyces cerevisiae yeast strains. Despite the potential of non-Saccharomyces strains for alcoholic fermentation, few studies have been performed at industrial level with these yeasts. Therefore, in this work, Agave tequilana juice was fermented at an industrial level using two non-Saccharomyces yeasts (Pichia kluyveri and Kluyveromyces marxianus) with fermentation efficiency higher than 85 %. Pichia kluyveri (GRO3) was more efficient for alcohol and ethyl lactate production than S. cerevisiae (AR5), while Kluyveromyces marxianus (GRO6) produced more isobutanol and ethyl-acetate than S. cerevisiae (AR5). The level of volatile compounds at the end of fermentation was compared with the tequila standard regulation. All volatile compounds were within the allowed range except for methanol, which was higher for S. cerevisiae (AR5) and K. marxianus (GRO6). The variations in methanol may have been caused by the Agave tequilana used for the tests, since this compound is not synthesized by these yeasts. PMID:23329062

  10. The roles of galactitol, galactose-1-phosphate, and phosphoglucomutase in galactose-induced toxicity in Saccharomyces cerevisiae.

    Science.gov (United States)

    de Jongh, Willem A; Bro, Christoffer; Ostergaard, Simon; Regenberg, Birgitte; Olsson, Lisbeth; Nielsen, Jens

    2008-10-01

    The uptake and catabolism of galactose by the yeast Saccharomyces cerevisiae is much lower than for glucose and fructose, and in applications of this yeast for utilization of complex substrates that contain galactose, for example, lignocellulose and raffinose, this causes prolonged fermentations. Galactose is metabolized via the Leloir pathway, and besides the industrial interest in improving the flux through this pathway it is also of medical relevance to study the Leloir pathway. Thus, genetic disorders in the genes encoding galactose-1-phosphate uridylyltransferase or galactokinase result in galactose toxicity both in patients with galactosemia and in yeast. In order to elucidate galactose related toxicity, which may explain the low uptake and catabolic rates of S. cerevisiae, we have studied the physiological characteristics and intracellular metabolite profiles of recombinant S. cerevisiae strains with improved or impaired growth on galactose. Aerobic batch cultivations on galactose of strains with different combinations of overexpression of the genes GAL1, GAL2, GAL7, and GAL10, which encode proteins that together convert extracellular galactose into glucose-1-phosphate, revealed a decrease in the maximum specific growth rate when compared to the reference strain. The hypothesized toxic intermediate galactose-1-phosphate cannot be the sole cause of galactose related toxicity, but indications were found that galactose-1-phosphate might cause a negative effect through inhibition of phosphoglucomutase. Furthermore, we show that galactitol is formed in S. cerevisiae, and that the combination of elevated intracellular galactitol concentration, and the ratio between galactose-1-phosphate concentration and phosphoglucomutase activity seems to be important for galactose related toxicity causing decreased growth rates. PMID:18421797

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

    OpenAIRE

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

    2008-01-01

    Abstract 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 relief of glucose limitation. Results The...

  12. Transcriptome-Based Characterization of Interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus in Lactose-Grown Chemostat Cocultures

    OpenAIRE

    Mendes, Filipa; Sieuwerts, Sander; Hulster, Erik; Almering, Marinka J. H.; Luttik, Marijke A. H.; Pronk, Jack T.; Smid, Eddy J.; Bron, Peter A.; Daran-lapujade, Pascale

    2013-01-01

    Mixed populations of Saccharomyces cerevisiae yeasts and lactic acid bacteria occur in many dairy, food, and beverage fermentations, but knowledge about their interactions is incomplete. In the present study, interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus, two microorganisms that co-occur in kefir fermentations, were studied during anaerobic growth on lactose. By combining physiological and transcriptome analysis of the two strains in the cocultu...

  13. The Snf1 Protein Kinase in the Yeast Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Usaite, Renata

    2008-01-01

    In yeast, Saccharomyces cerevisiae, the Snf1 protein kinase is primarily known as a key component of the glucose repression regulatory cascade. The Snf1 kinase is highly conserved among eukaryotes and its mammalian homolog AMPK is responsible for energy homeostasis in cells, organs and whole bodies. Failure in the AMPK regulatory cascade leads to metabolic disorders, such as obesity or type 2 diabetes. The knowledge about the Snf1 protein kinase remains to be of much interest in studying yeast carbon metabolism and human biology. To investigate the effect of Snf1 kinase and its regulatory subunit Snf4 on the regulation of glucose and galactose metabolism, I physiologically characterized ?snf1, ?snf4, and ?snf1?snf4 CEN.PK background yeast strains in glucose and glucose-galactose mixture batch cultivations (chapter 2). The results of this study showed that delayed induction of galactose catabolism was SNF1 or SNF4 gene deletion specific. In comparison to the reference strain, growth delay on galactose was found to last 2.4 times (7 hours) longer for the ?snf4, 3.1 times (10.5 hours) longer for the ?snf1, and 9.6 times (43 hours) longer for the ?snf1?snf4 strains. The maximum specific growth rates on galactose were found to be two to three times lower for the recombinant strains compared to the reference strain (0.13 h-1) and were found to be 0.07 h-1 for the ?snf1, 0.08 h-1 for the ?snf4 and 0.04 h-1 for the ?snf1?snf4 strain. In contrast to what is generally believed, the study showed that the Snf1 kinase was not solely responsible for the derepression of galactose metabolism. To investigate the regulatory role of Snf1 kinase on a global scale, the global scale mRNA, large-scale yeast quantitative proteome and metabolome datasets were generated. One of the largest yeast global quantitative proteome datasets (2388 proteins) to date was generated using Multidimensional Protein Identification Technology followed by quantitation using stable isotope labeling approach (chapter 3). The stable isotope labeling was compared to the spectral counting quantitative approach and the study showed that the stable isotope labeling approach is highly reproducible among biological replicates when complex protein mixtures containing small expression changes were analyzed. Where poor correlation between stable isotope labeling and spectral counting was found, the major reason behind the discrepancy was the lack of reproducible sampling for proteins with low spectral counts. To reconstruct a regulatory map of the yeast Snf1 protein kinase, I used the abundances of 5716 mRNAs, 2388 proteins, and 44 metabolites measured for the wild-type, ?snf1, ?snf4, and ?snf1?snf4 strains. By integrating these measurements with global protein-protein-interactions, protein-DNAinteractions and a genome-scale metabolic model, I mapped the complete network of interactions around the protein kinase Snf1 (chapters 4, 5). Through these interactions, I identified how the Snf1 protein kinase regulated cellular metabolism on gene or protein level. The study revealed that the Snf1 protein kinase played a far more extensive role in controlling both carbon and energy metabolism than previously anticipated. Similar to the function of AMPK in humans, my findings showed that Snf1 was a low energy checkpoint. Our results suggested that it was possible to use yeast more extensively as a model system for studying the molecular mechanisms underlying the global regulation of AMPK in mammals.

  14. Unraveling condition specific gene transcriptional regulatory networks in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Kluger Yuval

    2006-03-01

    Full Text Available Abstract 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] Additional File 1 overview of supplemental data Click here for file Additional File 2 experimental conditions for each link in figure 5. These are the experimental conditions in which the links are likely to be active. Click here for file Additional File 3 experimental conditions for each link in figure 7. These are the experimental conditions in which the links are likely to be active. Click here for file Additional File 4 Alon's transcriptional regulatory sub-network. Sparse representation of the Alon's network where column one represents the TF and column two represents the target. Entry of 1(2 corresponds to activation (suppression Click here for file Additional File 5 The Union of Alon's and Palsson's transcriptional regulatory sub-networks. Sparse representation of this unified network where column one represents the TF and column two represents the target. Entry of 1(2 corresponds to activation (suppression Click here for file Additional File 6 Predicted transcriptional regulatory links obtained by applying the LINK model to Alon's network. Each link is accompanied by a list of experiments in which it is likely to be functional. Click here for file Additional File 7 Predicted transcriptional regulatory links obtained by applying the LINK model to the combined network obtained by unifying Alon's and Palsson's networks. Each link is accompanied by a list of experiments in which it is likely to be functional. Click here for file Additional File 8 Predicted transcriptional regulatory links obtained by applying the STAR model to Alon's network. Each link is accompanied by a list of experiments in which it is likely to be functional. Click here for file Additional File 9 Predicted transcriptional regulatory links obtained by applying the STAR model to the combined network obtained by unifying Alon's and Palsson's networks. Each link is accompanied by a list of experiments in which it is likely to be functional. Click here for file Additional File 10 Exploring the parameter space. Overlap between our condition specific predicted networks and condition specific ChIP-on-chip data. Click here for file

  15. Evaluation of Brachypodium distachyon L-Tyrosine Decarboxylase Using L-Tyrosine Over-Producing Saccharomyces cerevisiae

    Science.gov (United States)

    Noda, Shuhei; Shirai, Tomokazu; Mochida, Keiichi; Matsuda, Fumio; Oyama, Sachiko; Okamoto, Mami; Kondo, Akihiko

    2015-01-01

    To demonstrate that herbaceous biomass is a versatile gene resource, we focused on the model plant Brachypodium distachyon, and screened the B. distachyon for homologs of tyrosine decarboxylase (TDC), which is involved in the modification of aromatic compounds. A total of 5 candidate genes were identified in cDNA libraries of B. distachyon and were introduced into Saccharomyces cerevisiae to evaluate TDC expression and tyramine production. It is suggested that two TDCs encoded in the transcripts Bradi2g51120.1 and Bradi2g51170.1 have L-tyrosine decarboxylation activity. Bradi2g51170.1 was introduced into the L-tyrosine over-producing strain of S. cerevisiae that was constructed by the introduction of mutant genes that promote deregulated feedback inhibition. The amount of tyramine produced by the resulting transformant was 6.6-fold higher (approximately 200 mg/L) than the control strain, indicating that B. distachyon TDC effectively converts L-tyrosine to tyramine. Our results suggest that B. distachyon possesses enzymes that are capable of modifying aromatic residues, and that S. cerevisiae is a suitable host for the production of L-tyrosine derivatives. PMID:25996877

  16. Impact of overexpressing NADH kinase on glucose and xylose metabolism in recombinant xylose-utilizing Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Hou, Jin; Vemuri, G. N.

    2009-01-01

    During growth of Saccharomyces cerevisiae on glucose, the redox cofactors NADH and NADPH are predominantly involved in catabolism and biosynthesis, respectively. A deviation from the optimal level of these cofactors often results in major changes in the substrate uptake and biomass formation. However, the metabolism of xylose by recombinant S. cerevisiae carrying xylose reductase and xylitol dehydrogenase from the fungal pathway requires both NADH and NADPH and creates cofactor imbalance during growth on xylose. As one possible solution to overcoming this imbalance, the effect of overexpressing the native NADH kinase (encoded by the POS5 gene) in xylose-consuming recombinant S. cerevisiae directed either into the cytosol or to the mitochondria was evaluated. The physiology of the NADH kinase containing strains was also evaluated during growth on glucose. Overexpressing NADH kinase in the cytosol redirected carbon flow from CO2 to ethanol during aerobic growth on glucose and to ethanol and acetate during anaerobic growth on glucose. However, cytosolic NADH kinase has an opposite effect during anaerobic metabolism of xylose consumption by channeling carbon flow from ethanol to xylitol. In contrast, overexpressing NADH kinase in the mitochondria did not affect the physiology to a large extent. Overall, although NADH kinase did not increase the rate of xylose consumption, we believe that it can provide an important source of NADPH in yeast, which can be useful for metabolic engineering strategies where the redox fluxes are manipulated.

  17. Tomato QM-like protein protects Saccharomyces cerevisiae cells against oxidative stress by regulating intracellular proline levels.

    Science.gov (United States)

    Chen, Changbin; Wanduragala, Srimevan; Becker, Donald F; Dickman, Martin B

    2006-06-01

    Exogenous proline can protect cells of Saccharomyces cerevisiae from oxidative stress. We altered intracellular proline levels by overexpressing the proline dehydrogenase gene (PUT1) of S. cerevisiae. Put1p performs the first enzymatic step of proline degradation in S. cerevisiae. Overexpression of Put1p results in low proline levels and hypersensitivity to oxidants, such as hydrogen peroxide and paraquat. A put1-disrupted yeast mutant deficient in Put1p activity has increased protection from oxidative stress and increased proline levels. Following a conditional life/death screen in yeast, we identified a tomato (Lycopersicon esculentum) gene encoding a QM-like protein (tQM) and found that stable expression of tQM in the Put1p-overexpressing strain conferred protection against oxidative damage from H2O2, paraquat, and heat. This protection was correlated with reactive oxygen species (ROS) reduction and increased proline accumulation. A yeast two-hybrid system assay was used to show that tQM physically interacts with Put1p in yeast, suggesting that tQM is directly involved in modulating proline levels. tQM also can rescue yeast from the lethality mediated by the mammalian proapoptotic protein Bax, through the inhibition of ROS generation. Our results suggest that tQM is a component of various stress response pathways and may function in proline-mediated stress tolerance in plants. PMID:16751508

  18. Saccharomyces paradoxus and Saccharomyces cerevisiae reside on oak trees in New Zealand: evidence for migration from Europe and interspecies hybrids.

    Science.gov (United States)

    Zhang, Hanyao; Skelton, Aaron; Gardner, Richard C; Goddard, Matthew R

    2010-11-01

    Saccharomyces cerevisiae and Saccharomyces paradoxus are used as model systems for molecular, cell and evolutionary biology; yet we know comparatively little of their ecology. One niche from which these species have been isolated is oak bark. There are no reports of these species from oak in the Southern Hemisphere. We describe the recovery of both S. cerevisiae and S. paradoxus from oak in New Zealand (NZ), and provide evidence for introgression between the species. Genetic inference shows that the oak S. cerevisiae are closely related to strains isolated from NZ and Australian vineyards, but that the S. paradoxus strains are very closely related to European isolates. This discovery is surprising as the current model of S. paradoxus biogeography suggests that global dispersal is rare. We test one idea to explain how members of the European S. paradoxus population might come to be in NZ: they were transported here along with acorns brought by migrants approximately 200 years ago. We show that S. paradoxus is associated with acorns and thus provide a potential mechanism for the unwitting global dispersal of S. paradoxus by humans. PMID:20868381

  19. Transcriptional responses to glucose in Saccharomyces cerevisiae strains lacking a functional protein kinase A

    Directory of Open Access Journals (Sweden)

    Livas Daniela

    2011-08-01

    Full Text Available Abstract Background The pattern of gene transcripts in the yeast Saccharomyces cerevisiae is strongly affected by the presence of glucose. An increased activity of protein kinase A (PKA, triggered by a rise in the intracellular concentration of cAMP, can account for many of the effects of glucose on transcription. In S. cerevisiae three genes, TPK1, TPK2, and TPK3, encode catalytic subunits of PKA. The lack of viability of tpk1 tpk2 tpk3 triple mutants may be suppressed by mutations such as yak1 or msn2/msn4. To investigate the requirement for PKA in glucose control of gene expression, we have compared the effects of glucose on global transcription in a wild-type strain and in two strains devoid of PKA activity, tpk1 tpk2 tpk3 yak1 and tpk1 tpk2 tpk3 msn2 msn4. Results We have identified different classes of genes that can be induced -or repressed- by glucose in the absence of PKA. Representative examples are genes required for glucose utilization and genes involved in the metabolism of other carbon sources, respectively. Among the genes responding to glucose in strains devoid of PKA some are also controlled by a redundant signalling pathway involving PKA activation, while others are not affected when PKA is activated through an increase in cAMP concentration. On the other hand, among genes that do not respond to glucose in the absence of PKA, some give a full response to increased cAMP levels, even in the absence of glucose, while others appear to require the cooperation of different signalling pathways. We show also that, for a number of genes controlled by glucose through a PKA-dependent pathway, the changes in mRNA levels are transient. We found that, in cells grown in gluconeogenic conditions, expression of a small number of genes, mainly connected with the response to stress, is reduced in the strains lacking PKA. Conclusions In S. cerevisiae, the transcriptional responses to glucose are triggered by a variety of pathways, alone or in combination, in which PKA is often involved. Redundant signalling pathways confer a greater robustness to the response to glucose, while cooperative pathways provide a greater flexibility.

  20. Comportamento celular e resposta antioxidante diferenciados de Saccharomyces cerevisiae e de Saccharomyces chevalieri ao metavanadato de amónio / Different cellular behaviour and antioxidant response of Saccharomyces cerevisiae and Saccharomyces chevalieri growing in presence of ammonium metavanadate

    Scientific Electronic Library Online (English)

    R., Ferreira; I., Alves-Pereira; S., Magriço; C., Ferraz-Franco.

    2007-01-01

    Full Text Available A fermentação do vinho é um processo microbiológico complexo que requere a presença de leveduras adaptadas a condições de stresse. No ambiente celular de organismos aeróbios ocorrem naturalmente espécies reactivas de oxigénio (ROS) como subprodutos da respiração mitocondrial. A elevada reactividade [...] destas espécies químicas pode gerar danos moleculares que, em alguns casos, levam à morte celular. Em condições fisiológicas normais ou como resposta ao stresse oxidativo, a célula pode desencadear respostas adaptativas que envolvem mecanismos antioxidantes como os enzimas glutationo redutase (GR; EC 1.6.4.2) e catalases T (CAT T; EC 1.11.1.6) e A (CAT A; EC 1.11.1.6). O vanádio, um metal pesado presente em alguns fitofármacos, pode também com portar-se como um gerador de ROS, alterando o estado redox intracelular e exercendo efeitos nocivos em leveduras expostas a quantidade excessiva deste elemento. O principal objectivo deste trabalho foi comparar o efeito do metavanadato de amónio (NH4VO3), um sal pentavalente de vanádio, na viabilidade celular e nas actividades enzimáticas GR, CAT T e CAT A das leveduras vínicas Saccharomyces cerevisiae UE-ME3 e Saccharomyces chevalieri UE-ME1. Os resultados obtidos mostram que S. chevalieri UE-ME1 revelou menor tolerância ao NH4VO3 do que S. cerevisiae UE-ME3, uma vez que culturas de S. chevalieri não sobreviveram para valores de concentração do sal de vanádio superiores a 7,5 mM enquanto que células de S. cerevisiae mantiveram-se viáveis em presença de metavanadato de amónio 75 mM. As actividades enzimáticas estudadas apresentaram em S. chevalieri valores muito inferiores aos que foram determinados em S. cerevisiae embora em ambas as espécies de levedura o NH4VO3 pareça comportarse como um indutor de stresse oxidativo ao provocar um decréscimo significativo da actividade GR (P Abstract in english The fermentation of wine is a complex microbiological process which requires yeast adaptation to stress conditions. In the cellular environment of aerobic organisms naturally reactive oxygen species (ROS) occurs as by-products of mitochondrial respiration. The higher reactivity of these chemical spe [...] cies could cause molecular damages that in several cases induce cellular death. In common physiological conditions or as response to oxidative stress, the cell can generate adapted responses which involve antioxidants mechanisms as glutathione reductase (GR; EC 1.6.4.2) and catalase T (CAT T; EC 1.11.1.6) and A (CAT A; EC 1.11.1.6) enzymes. Vanadium, a heavy metal present in several pesticides could generate ROS changing the intracellular redox state and cause deleterious effects in yeasts exposed to higher levels of this element. The main objective of this work was to compare the effects of ammonium metavanadate (NH4VO3), a pentavalent salt of vanadium on cellular viability and GR, CAT T and CAT A activities of wine yeast Saccharomyces cerevisiae UE-ME3 and Saccharomyces chevalieri UE-ME1. The results obtained show that S. chevalieri UE-ME1 has lower tolerance to NH4VO3 than S. cerevisiae UE-ME3, since S. chevalieri cultures do not survive to concentration values of ammonium metavanadate higher than 7,5 mM, whereas S. cerevisiae cells are still viable in the presence of 75 mM. S. chevalieri has an enzymatic activity lower than S. cerevisiae, although for both yeast species NH4VO3 could behave as oxidative stress inductor, causing a significant decrease of GR activity (P

  1. Two independent activities define Ccm1p as a moonlighting protein in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    J. Ignacio Moreno

    2012-09-01

    Full Text Available Ccm1p is a nuclear-encoded PPR (pentatricopeptide repeat protein that localizes into mitochondria of Saccharomyces cerevisiae. It was first defined as an essential factor to remove the bI4 [COB (cytochrome b fourth intron] and aI4 [COX1 (cytochrome c oxidase subunit 1 fourth intron] of pre-mRNAs, along with bI4 maturase, a protein encoded by part of bI4 and preceding exons that removes the intronic RNA sequence that codes for it. Later on, Ccm1p was described as key to maintain the steady-state levels of the mitoribosome small subunit RNA (15S rRNA. bI4 maturase is produced inside the mitochondria and therefore its activity depends on the functionality of mitochondrial translation. This report addresses the dilemma of whether Ccm1p supports bI4 maturase activity by keeping steady-state levels of 15S rRNA or separately and directly supports bI4 maturase activity per se. Experiments involving loss of Ccm1p, SMDC (sudden mitochondrial deprivation of Ccm1p and mutations in one of the PPR (pentatricopeptide repeat motifs revealed that the failure of bI4 maturase activity in CCM1 deletion mutants was not due to a malfunction of the translational machinery. Both functions were found to be independent, defining Ccm1p as a moonlighting protein. bI4 maturase activity was significantly more dependent on Ccm1p levels than the maintenance of 15S rRNA. The novel strategy of SMDC described here allowed the study of immediate short-term effects, before the mutant phenotype was definitively established. This approach can be also applied for further studies on 15S rRNA stability and mitoribosome assembly.

  2. Production of extracellular and total invertase by Candida utilis, Saccharomyces cerevisiae, and other yeasts.

    Science.gov (United States)

    DWORSCHACK, R G; WICKERHAM, L J

    1961-07-01

    Some strains of Candida utilis produce exceptionally large amounts of extracellular and total invertase. Strain Y-900 of C. utilis produces high yields whether the carbon source is sucrose, glucose, maltose, or xylose and still higher yields with lactic acid, glycerol, and ethyl alcohol. Approximately 20 to 30% of the total invertase of C. utilis is extracellular. Strains of Saccharomyces cerevisiae and Saccharomyces carlsbergensis are generally inferior to C. utilis in production of extracellular and total invertase, the difference being accentuated in shaken cultures. The industrial yeasts are generally superior in invertase production to the other yeasts included in the survey. PMID:13725351

  3. Introduction of extra telomeric DNA sequences into Saccharomyces cerevisiae results in telomere elongation.

    OpenAIRE

    Runge, K W; Zakian, V A

    1989-01-01

    The termini of Saccharomyces cerevisiae chromosomes consist of tracts of C1-3A (one to three cytosine and one adenine residue) sequences of approximately 450 base pairs in length. To gain insights into trans-acting factors at telomeres, high-copy-number linear and circular plasmids containing tracts of C1-3A sequences were introduced into S. cerevisiae. We devised a novel system to distinguish by color colonies that maintained the vector at 1 to 5, 20 to 50, and 100 to 400 copies per cell and...

  4. Hsp42 is the general small heat shock protein in the cytosol of Saccharomyces cerevisiae

    OpenAIRE

    Haslbeck, Martin; Braun, Nathalie; Stromer, Thusnelda; Richter, Bettina; Model, Natascha; Weinkauf, Sevil; Buchner, Johannes

    2004-01-01

    Small heat shock proteins (sHsps) are ubiquitous molecular chaperones that prevent the unspecific aggregation of proteins. So far, Hsp26 was the only unambiguously identified member of the sHsp family in Saccharomyces cerevisiae. We show here that the sHsp system in the cytosol of S. cerevisiae consists of two proteins, Hsp26 and Hsp42. Hsp42 forms large dynamic oligomers with a barrel-like structure. In contrast to Hsp26, which functions predominantly at heat shock temperatures, Hsp42 is act...

  5. Advances in metabolic engineering of yeast Saccharomyces cerevisiae for production of chemicals

    DEFF Research Database (Denmark)

    Borodina, Irina; Nielsen, Jens

    2014-01-01

    Yeast Saccharomyces cerevisiae is an important industrial host for production of enzymes, pharmaceutical and nutraceutical ingredients and recently also commodity chemicals and biofuels. Here, we review the advances in modeling and synthetic biology tools and how these tools can speed up the development of yeast cell factories. We also present an overview of metabolic engineering strategies for developing yeast strains for production of polymer monomers: lactic, succinic, and cis,cis-muconic acids. S. cerevisiae has already firmly established itself as a cell factory in industrial biotechnology and the advances in yeast strain engineering will stimulate development of novel yeast-based processes for chemicals production.

  6. Integrated phospholipidomics and transcriptomics analysis of Saccharomyces cerevisiae with enhanced tolerance to a mixture of acetic acid, furfural, and phenol

    Science.gov (United States)

    A mixture of acetic acid, furfural and phenol (AFP), three representative lignocellulose derived inhibitors, significantly inhibited the growth and bioethanol production of Saccharomyces cerevisiae. In order to uncover mechanisms behind the enhanced tolerance of an inhibitor-tolerant S.cerevisiae s...

  7. O emprego de fermento de pão, Saccharomyces cerevisiae, na síntese de feromônios / Baker's yeast, Saccharomyces cerevisiae, as a tool for the synthesis of pheromones

    Scientific Electronic Library Online (English)

    Patrícia T., Baraldi; Arlene G., Corrêa.

    2004-06-01

    Full Text Available [...] Abstract in english The use of pheromones in integrated pest management has been increasing in the last years due to environmental concern. This development is accompanied by the search for simple, efficient and less aggressive synthetic methodologies for the preparation of pheromones. One of these methodologies includ [...] es microbiological reactions, more specifically biocatalytic reduction of carbonyl compounds using baker's yeast (Saccharomyces cerevisiae). This review presents the use of baker's yeast as an easy and cheap alternative to obtain enantiomerically enriched compounds employed in the synthesis of pheromones.

  8. ACÚMULO DE CÁDMIO POR Saccharomyces cerevisiae FERMENTANDO MOSTO DE CALDO DE CANA ACCUMULATION OF CADMIUM BY Saccharomyces cerevisiae FERMENTING MUST OF SUGAR-CANE

    OpenAIRE

    S.M.G. da SILVA; L.G. do Prado-Filho

    1998-01-01

    O presente trabalho estudou o acúmulo de cádmio (Cd) por Saccharomyces cerevisiae, fermentando mosto de caldo de cana com contaminações controladas, em níveis sub-tóxicos, do citado metal. O ensaio de fermentação foi conduzido em erlenmayers de 500 mL, acondicionados em estufa B.O.D. O mosto, não esterilizado, continha 12% de açúcares redutores totais (ART) e pH 4,5. Para a contaminação controlada empregou-se cloreto de cádmio em quatro níveis de contaminações: 0,5; 1,0; 2,0 ...

  9. A novel member of the split betaalphabeta fold: Solution structure of the hypothetical protein YML108W from Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    As part of the Northeast Structural Genomics Consortium pilot project focused on small eukaryotic proteins and protein domains, we have determined the NMR structure of the protein encoded by open reading frame YML108W from Saccharomyces cerevisiae. YML108W belongs to one of the numerous structural proteomics targets whose biological function is unknown. Moreover, this protein does not have sequence similarity to any other protein. The NMR structure of YML108W consists of a four-stranded b-sheet with strand order 2143 and two a-helices, with an overall topology of bbabba. Strand b1 runs parallel to b4, and b2:b1 and b4:b3 pairs are arranged in an antiparallel fashion. While this fold belongs to the split bab family, it appears to be unique among this family; it is a novel arrangement of secondary structure, thereby expanding the universe of protein folds

  10. Effects of altered 5'-flanking sequences on the in vivo expression of a Saccharomyces cerevisiae tRNATyr gene.

    OpenAIRE

    Shaw, K. J.; Olson, M. V.

    1984-01-01

    Deletion mutations ending in the 5'-flanking sequences of the Saccharomyces cerevisiae SUP4-o gene have been analyzed for their effects on gene expression. This ochre-suppressing tRNATyr gene was cloned into a S. cerevisiae centromeric plasmid, and its level of in vivo expression was monitored by observing the suppressor phenotype of the gene after transformation into S. cerevisiae. A deletion mutant that retains only four base pairs of the 5'-flanking sequence is profoundly deficient in expr...

  11. The Reacquisition of Biotin Prototrophy in Saccharomyces cerevisiae Involved Horizontal Gene Transfer, Gene Duplication and Gene Clustering

    OpenAIRE

    Hall, Charles; Dietrich, Fred S

    2007-01-01

    The synthesis of biotin, a vitamin required for many carboxylation reactions, is a variable trait in Saccharomyces cerevisiae. Many S. cerevisiae strains, including common laboratory strains, contain only a partial biotin synthesis pathway. We here report the identification of the first step necessary for the biotin synthesis pathway in S. cerevisiae. The biotin auxotroph strain S288c was able to grow on media lacking biotin when BIO1 and the known biotin synthesis gene BIO6 were introduced t...

  12. Co-utilization of L-arabinose and D-xylose by laboratory and industrial Saccharomyces cerevisiae strains

    OpenAIRE

    Boles Eckhard; Hahn-Hägerdal Bärbel; Wiedemann Beate; Karhumaa Kaisa; Gorwa-Grauslund Marie-F

    2006-01-01

    Abstract Background Fermentation of lignocellulosic biomass is an attractive alternative for the production of bioethanol. Traditionally, the yeast Saccharomyces cerevisiae is used in industrial ethanol fermentations. However, S. cerevisiae is naturally not able to ferment the pentose sugars D-xylose and L-arabinose, which are present in high amounts in lignocellulosic raw materials. Results We describe the engineering of laboratory and industrial S. cerevisiae strains to co-ferment the pento...

  13. Tomato QM-Like Protein Protects Saccharomyces cerevisiae Cells against Oxidative Stress by Regulating Intracellular Proline Levels

    OpenAIRE

    Chen, Changbin; Wanduragala, Srimevan; Becker, Donald F.; Dickman, Martin B.

    2006-01-01

    Exogenous proline can protect cells of Saccharomyces cerevisiae from oxidative stress. We altered intracellular proline levels by overexpressing the proline dehydrogenase gene (PUT1) of S. cerevisiae. Put1p performs the first enzymatic step of proline degradation in S. cerevisiae. Overexpression of Put1p results in low proline levels and hypersensitivity to oxidants, such as hydrogen peroxide and paraquat. A put1-disrupted yeast mutant deficient in Put1p activity has increased protection from...

  14. Development on ethanol production from xylose by recombinant Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Jin-Ying Yang

    2009-12-01

    Full Text Available Xylose is the second major fermentable sugar present in lignocellulosic hydrolysates, so its fermentation is essential for the economic con- version of lignocellulose to ethanol. However, the traditional ethanol production strain Sacch- aromyces cerevisiae does not naturally use xy-lose as a substrate. A number of different ap-proaches have been used to engineer yeasts to reconstruct the gene background of S. cerevi- siae in recent years. The recombinant strains showed better xylose fermentation quality by comparison with the natural strains. This review examines the research on S. cerevisiae strains that have been genetically modified or adapted to ferment xylose to ethanol from three aspects including construction of xylose transportation, xylose-metabolic pathway and inhibitor toler-ance improvement of S. cerevisiae.

  15. Transcriptional regulation of an hsp70 heat shock gene in the yeast Saccharomyces cerevisiae.

    OpenAIRE

    Slater, M. R.; Craig, E. A.

    1987-01-01

    The yeast Saccharomyces cerevisiae contains three heat-inducible hsp70 genes. We have characterized the promoter region of the hsp70 heat shock gene YG100, that also displays a basal level of expression. Deletion of the distal region of the promoter resulted in an 80% drop in the basal level of expression without affecting expression after heat shock. Progressive-deletion analysis suggested that sequences necessary for heat-inducible expression are more proximal, within 233 base pairs of the ...

  16. Monoubiquitination Is Sufficient To Signal Internalization of the Maltose Transporter in Saccharomyces cerevisiae

    OpenAIRE

    Lucero, Pilar; Peñalver, Élida; Vela, Laura; Lagunas, Rosario

    2000-01-01

    Monoubiquitination of the 12-transmembrane segment (12-TMS) Saccharomyces cerevisiae maltose transporter promoted the maximal internalization rate of this protein. This modification is similar to that of the 7-TMS ?-factor receptor but different from that of the 12-TMS uracil and general amino acid permeases. This result shows that binding of ubiquitin-Lys63 chains is not required for maximal internalization of all 12-TMS-containing proteins.

  17. Dynamics of Glycolytic Regulation during Adaptation of Saccharomyces cerevisiae to Fermentative Metabolism? †

    OpenAIRE

    Van Den Brink, Joost; Canelas, André B.; van Gulik, Walter M.; Pronk, Jack T; Heijnen, Joseph J; de Winde, Johannes H.; Daran-Lapujade, Pascale

    2008-01-01

    The ability of baker's yeast (Saccharomyces cerevisiae) to rapidly increase its glycolytic flux upon a switch from respiratory to fermentative sugar metabolism is an important characteristic for many of its multiple industrial applications. An increased glycolytic flux can be achieved by an increase in the glycolytic enzyme capacities (Vmax) and/or by changes in the concentrations of low-molecular-weight substrates, products, and effectors. The goal of the present study was to understand the ...

  18. Extreme calorie restriction and energy source starvation in Saccharomyces cerevisiae represent distinct physiological states:

    OpenAIRE

    Boender, L.G.M.; Almering, M J H; Dijk, M. van; Maris, A.J.A., van; Winde, J.H. de; Pronk, J T; Daran-Lapujade, P

    2011-01-01

    Cultivation methods used to investigate microbial calorie restriction often result in carbon and energy starvation. This study aims to dissect cellular responses to calorie restriction and starvation in Saccharomyces cerevisiae by using retentostat cultivation. In retentostats, cells are continuously supplied with a small, constant carbon and energy supply, sufficient for maintenance of cellular viability and integrity but insufficient for growth. When glucose-limited retentostats cultivated ...

  19. The sexual inducer of Volvox carteri. Its large-scale production and secretion by Saccharomyces cerevisiae

    OpenAIRE

    Haas, E.; Sumper, Manfred

    1991-01-01

    The DNA sequence coding for the sexual inducer glycoprotein of Volvox carteri and its N-terminal signal peptide was placed under the control of the repressible acid phosphatase promoter of the yeast Saccharomyces cerevisiae in a yeast-E. coli shuttle vector. Yeast transformed by this construct synthesized and secreted into the culture medium biologically active inducer in amounts two to three orders of magnitude higher than observed in the Volvox system.

  20. Ethanol fermentation of a diluted molasses medium by Saccharomyces cerevisiae immobilized on chrysotile

    OpenAIRE

    Monte Alegre Ranulfo; Rigo Maurício; Joekes Inés

    2003-01-01

    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 CO2 production increased roughly 27 % during the first 30 minutes, compared to systems containing no chrysotile. A study of continuous alcoholic fermentation wit...

  1. Saccharomyces cerevisiae has distinct adaptive responses to both hydrogen peroxide and menadione.

    OpenAIRE

    Jamieson, D. J.

    1992-01-01

    Treatment of Saccharomyces cerevisiae cells with low concentrations of either hydrogen peroxide or menadione (a superoxide-generating agent) induces adaptive responses which protect cells from the lethal effects of subsequent challenge with higher concentrations of these oxidants. Pretreatment with menadione is protective against cell killing by hydrogen peroxide; however, pretreatment with hydrogen peroxide is unable to protect cells from subsequent challenge with menadione. This suggests th...

  2. Carboxypeptidase S- and Carboxypeptidase Y-Deficient Mutants of Saccharomyces cerevisiae

    OpenAIRE

    Wolf, Dieter H.; Ehmann, Claudia

    1981-01-01

    A new carboxypeptidase (carboxypeptidase S) was found in a Saccharomyces cerevisiae strain lacking carboxypeptidase Y (D. H. Wolf and U. Weiser, Eur. J. Biochem. 73:553-556, 1977). Mutants devoid of carboxypeptidase S activity were isolated from a mutant strain that was also deficient in carboxypeptidase Y. Four mutants were analyzed in detail and fell into one complementation group. The defect segregated 2:2 in meiotic tetrads. Gene dosage experiments indicated that the mutation might reside...

  3. Predicción del contenido intracelular de trehalosa en el proceso de producción de biomasa de Saccharomyces cerevisiae

    OpenAIRE

    Aranda, J. S.; Cabrera, A. I.; Chairez, J. I.

    2008-01-01

    La trehalosa es un carbohidrato dimérico componente de la levadura de panificación Saccharomyces cerevisiae, y es considerado como indicador de la capacidad fermentativa y de la viabilidad de las células. En procesos de producción de levadura, se busca inducir una acumulación intracelular de trehalosa. Por ser un compuesto citoplásmico, la cuantificación de la trehalosa requiere de tomas de muestra y de métodos analíticos posteriores. Así, el conocimiento del contenido c...

  4. Stress Tolerance in Doughs of Saccharomyces cerevisiae Trehalase Mutants Derived from Commercial Baker’s Yeast

    OpenAIRE

    Shima, Jun; Hino, Akihiro; Yamada-Iyo, Chie; SUZUKI, Yasuo; Nakajima, Ryouichi; Watanabe, Hajime; Mori, Katsumi; Takano, Hiroyuki

    1999-01-01

    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 (?nth1), acid trehalase mutants (?ath1), and double mutants (?nth1 ath1) by using commercial baker’s yeast strains as the parent strains and the gene disruption me...

  5. Genome-Wide Analysis of Sterol-Lipid Storage and Trafficking in Saccharomyces cerevisiae?

    OpenAIRE

    Fei, Weihua; Alfaro, Gabriel; Muthusamy, Baby-Periyanayaki; Klaassen, Zachary; Graham, Todd R.; Yang, Hongyuan; Beh, Christopher T.

    2007-01-01

    The pandemic of lipid-related disease necessitates a determination of how cholesterol and other lipids are transported and stored within cells. The first step in this determination is the identification of the genes involved in these transport and storage processes. Using genome-wide screens, we identified 56 yeast (Saccharomyces cerevisiae) genes involved in sterol-lipid biosynthesis, intracellular trafficking, and/or neutral-lipid storage. Direct biochemical and cytological examination of m...

  6. Protective effect of acetic acid against ethanol-induced cell death in "Saccharomyces cerevisiae"

    OpenAIRE

    Afonso, Andreia Fernandes

    2011-01-01

    O etanol é um produto final bem conhecido da fermentação alcoólica realizada por Saccharomyces cerevisiae. Em altas concentrações, é responsável pela redução de viabilidade celular e inibição da fermentação. Além disso, durante a fermentação alguns ácidos fracos, como os ácidos acético, butírico e pirúvico, produzidos pelo metabolismo da levedura, podem acumular-se no meio de crescimento e aumentar a toxicidade do etanol, o que resulta numa maior inibição de cresciment...

  7. A New Purple Fluorescent Color Marker for Genetic Studies in Saccharomyces cerevisiae and Candida albicans

    OpenAIRE

    Keppler-Ross, Sabine; Noffz, Christine; Dean, Neta

    2008-01-01

    The ability to visualize cellular events by linking them to color or fluorescence changes has been an invaluable tool for biology. We describe a novel plasmid-borne color marker whose expression in yeast leads to purple-colored cells that are also brightly fluorescent. This dominant marker provides a useful tool for rapidly screening plasmid maintenance using a visual or fluorescence assay in both Saccharomyces cerevisiae and Candida albicans.

  8. Sensing Inorganic Phosphate Starvation by the Phosphate-Responsive (PHO) Signaling Pathway of Saccharomyces cerevisiae

    OpenAIRE

    Choi, Joonhyuk

    2013-01-01

    Inorganic phosphate \\((P_i)\\) is an essential nutrient whose intracellular levels are maintained by the PHO pathway in Saccharomyces cerevisiae. \\(P_i\\) limitation triggers upregulation of the PHO genes whose gene products primarily function to counterbalance the \\(P_i\\) deficiency. Despite a growing catalogue of genes that are involved in signaling of the PHO pathway, little is known about how cells actually sense \\(P_i\\) limitation. To better characterize the \\(P_i\\) sensing mechanism, I ex...

  9. Microbial transformations of ferulic acid by Saccharomyces cerevisiae and Pseudomonas fluorescens.

    OpenAIRE

    Huang, Z.; Dostal, L.; Rosazza, J. P.

    1993-01-01

    Saccharomyces cerevisiae (dry baker's yeast) and Pseudomonas fluorescens were used to convert trans-ferulic acid into 4-hydroxy-3-methoxystyrene in 96 and 89% yields, respectively. The metabolites were isolated by solid-phase extraction and analyzed by thin-layer chromatography and high-performance liquid chromatography. The identities of the metabolites were determined by 1H- and 13C-nuclear magnetic resonance spectroscopy and by mass spectrometry. The mechanism of the decarboxylation of fer...

  10. X-Ray Absorption Spectroscopy of Cuprous-Thiolate Clusters in Saccharomyces cerevisiae Metallothionein

    OpenAIRE

    Zhang, Limei; Pickering, Ingrid J; Winge, Dennis R.; George, Graham N.

    2008-01-01

    Copper (Cu) metallothioneins are cuprous-thiolate proteins that contain multimetallic clusters, and are thought to have dual functions of Cu storage and Cu detoxification. We have used a combination of X-ray absorption spectroscopy (XAS) and density-functional theory (DFT) to investigate the nature of Cu binding to Saccharomyces cerevisiae metallothionein. We found that the XAS of metallothionein prepared, containing a full complement of Cu, was quantitatively consistent with the crystal stru...

  11. Effects of mill stream flours technological quality on fermentative activity of baker's yeast Saccharomyces cerevisiae

    OpenAIRE

    Miri? Katarina V.; Pejin Dušanka J.

    2008-01-01

    This work in concerned with the interdependence between technological quality of mill stream flours and fermentative activity of baker's yeast Saccharomyces cerevisiae. Each mill stream flour has its own specific properties, determined by the particle size, technological phase of its formation and part of the wheat kernel it consists of. Biochemical complexity of dough during examination of fermentative activity of baker's yeast confirmed the influence of a number of physical and biochemical ...

  12. Cloning and Expression of a Schwanniomyces occidentalis ?-Amylase Gene in Saccharomyces cerevisiae

    OpenAIRE

    Wang, Tsung Tsan; Lin, Long Liu; Hsu, Wen Hwei

    1989-01-01

    An ?-amylase gene (AMY) was cloned from Schwanniomyces occidentalis CCRC 21164 into Saccharomyces cerevisiae AH22 by inserting Sau3AI-generated DNA fragments into the BamHI site of YEp16. The 5-kilobase insert was shown to direct the synthesis of ?-amylase. After subclones containing various lengths of restricted fragments were screened, a 3.4-kilobase fragment of the donor strain DNA was found to be sufficient for ?-amylase synthesis. The concentration of ?-amylase in culture broth produ...

  13. Regulation of hydrogen sulfide liberation in wine-producing Saccharomyces cerevisiae strains by assimilable nitrogen.

    OpenAIRE

    Jiranek, V.; Langridge, P.; Henschke, P. A.

    1995-01-01

    Saccharomyces cerevisiae wine-producing yeast cultures grown under model winemaking conditions could be induced to liberate hydrogen sulfide (H2S) by starvation for assimilable nitrogen. The amount of H2S produced was dependent on the yeast strain, the sulfur precursor compound, the culture growth rate, and the activity of the sulfite reductase enzyme (EC 1.8.1.2) immediately before nitrogen depletion. Increased H2S formation relative to its utilization by metabolism was not a consequence of ...

  14. Saccharomyces cerevisiae sec59 cells are deficient in dolichol kinase activity.

    OpenAIRE

    L. Heller; Orlean, P.; Adair, W L

    1992-01-01

    The temperature-sensitive Saccharomyces cerevisiae mutant sec59 accumulates inactive and incompletely glycosylated protein precursors in its endoplasmic reticulum at the restrictive temperature. O-mannosylation and glycosyl phosphatidylinositol membrane anchoring of protein are also abolished, consistent with a deficiency in dolichyl phosphate mannose. Membranes prepared from sec59 cells that had been shifted to the restrictive temperature, however, made normal amounts of dolichyl phosphate m...

  15. Characterization of synthetic DNA bar codes in Saccharomyces cerevisiae gene-deletion strains

    OpenAIRE

    Eason, Robert G.; Pourmand, Nader; Tongprasit, Waraporn; Herman, Zelek S.; Anthony, Kevin; Jejelowo, Olufisayo; Davis, Ronald W.; Stolc, Viktor

    2004-01-01

    Incorporation of strain-specific synthetic DNA tags into yeast Saccharomyces cerevisiae gene-deletion strains has enabled identification of gene functions by massively parallel growth rate analysis. However, it is important to confirm the sequences of these tags, because mutations introduced during construction could lead to significant errors in hybridization performance. To validate this experimental system, we sequenced 11,812 synthetic 20-mer molecular bar codes and adjacent sequences (>1...

  16. Role of inositol-containing sphingolipids in Saccharomyces cerevisiae during inositol starvation.

    OpenAIRE

    Hanson, B A

    1984-01-01

    The in vitro lipid requirements of UDP-N-acetylglucosamine-dolichol phosphate N-acetylglucosamine-1-phosphotransferase for the inositol-containing sphingolipids from Saccharomyces cerevisiae were characterized in terms of concentration and specificity. The effects of combinations of lipids, especially phosphatidylinositol and the inositol-containing sphingolipids, were also tested on the transferase. Phosphatidylinositol and phosphatidylglycerol stimulated the enzyme 3.3- and 2.8-fold, respec...

  17. Secretion can proceed uncoupled from net plasma membrane expansion in inositol-starved Saccharomyces cerevisiae.

    OpenAIRE

    Atkinson, K D; Ramirez, R M

    1984-01-01

    Secretion of acid phosphatase and invertase was examined in an inositol-requiring ino1 mutant of the yeast Saccharomyces cerevisiae. Inositol starvation is known to block plasma membrane expansion, presumably due to restricted membrane phospholipid synthesis. If membrane expansion and extracellular protein secretion are accomplished by the same intracellular transport process, one would expect secretion to fail coordinately with cessation of plasma membrane growth in inositol-starved cells. I...

  18. Metabolism of myo-inositol during sporulation of myo-inositol-requiring Saccharomyces cerevisiae.

    OpenAIRE

    Schroeder, R.; Breitenbach, M.

    1981-01-01

    We investigated the sporulation properties of a series of diploid Saccharomyces cerevisiae strains homozygous for inositol auxotrophic markers. The strains required different amounts of inositol for the completion of sporulation. Shift experiments revealed two phases of inositol requirement during sporulation which coincided with the two phases of lipid synthesis found by earlier workers. Phase I was at the beginning and during premeiotic deoxyribonucleic acid synthesis; phase II immediately ...

  19. Effect of Saccharomyces cerevisiae Fermentation Product on Lactation Performance and Lipopolysaccharide Concentration of Dairy Cows

    OpenAIRE

    Zhang, Rui-yang; Yoon, Ilkyu; Zhu, Wei-yun; Mao, Sheng-yong

    2013-01-01

    To evaluate lactation performance and changes in plasma and fecal lipopolysaccharide (LPS) concentrations in response to the supplementation of Saccharomyces cerevisiae fermentation product (SC), two dairy farms were selected. On each farm, 32 cows in early to mid lactation (21 to 140 DIM) were blocked by parity and days in milk (DIM), and randomly assigned to one of the two treatments within block (Control or 56 g SC/cow/d). Effect of SC on lactation performance (daily) and changes in blood ...

  20. Investigation of Two Distinct Flavone Synthases for Plant-Specific Flavone Biosynthesis in Saccharomyces cerevisiae

    OpenAIRE

    Leonard, Effendi; Yan, Yajun.; Lim, Kok Hong; Koffas, Mattheos A. G.

    2005-01-01

    Flavones are plant secondary metabolites that have wide pharmaceutical and nutraceutical applications. We previously constructed a recombinant flavanone pathway by expressing in Saccharomyces cerevisiae a four-step recombinant pathway that consists of cinnamate-4 hydroxylase, 4-coumaroyl:coenzyme A ligase, chalcone synthase, and chalcone isomerase. In the present work, the biosynthesis of flavones by two distinct flavone synthases was evaluated by introducing a soluble flavone synthase I (FSI...

  1. Lipid Raft-Based Membrane Compartmentation of a Plant Transport Protein Expressed in Saccharomyces cerevisiae

    OpenAIRE

    Grossmann, Guido; Opekarova, Miroslava; Novakova, Linda; Stolz, Jürgen; Tanner, Widmar

    2006-01-01

    The hexose-proton symporter HUP1 shows a spotty distribution in the plasma membrane of the green alga Chlorella kessleri. Chlorella cannot be transformed so far. To study the membrane localization of the HUP1 protein in detail, the symporter was fused to green fluorescent protein (GFP) and heterologously expressed in Saccharomyces cerevisiae and Schizosaccharomyces pombe. In these organisms, the HUP1 protein has previously been shown to be fully active. The GFP fusion protein was exclusively ...

  2. Statistics-based model for prediction of chemical biosynthesis yield from Saccharomyces cerevisiae

    OpenAIRE

    Leonard Effendi; Xiao Yi; Varman Arul M; Tang Yinjie J

    2011-01-01

    Abstract Background The robustness of Saccharomyces cerevisiae in facilitating industrial-scale production of ethanol extends its utilization as a platform to synthesize other metabolites. Metabolic engineering strategies, typically via pathway overexpression and deletion, continue to play a key role for optimizing the conversion efficiency of substrates into the desired products. However, chemical production titer or yield remains difficult to predict based on reaction stoichiometry and mass...

  3. Metabolic-Flux Profiling of the Yeasts Saccharomyces cerevisiae and Pichia stipitis

    OpenAIRE

    Fiaux, Jocelyne; C?akar, Z. Petek; Sonderegger, Marco; Wu?thrich, Kurt; Szyperski, Thomas; Sauer, Uwe

    2003-01-01

    The so far largely uncharacterized central carbon metabolism of the yeast Pichia stipitis was explored in batch and glucose-limited chemostat cultures using metabolic-flux ratio analysis by nuclear magnetic resonance. The concomitantly characterized network of active metabolic pathways was compared to those identified in Saccharomyces cerevisiae, which led to the following conclusions. (i) There is a remarkably low use of the non-oxidative pentose phosphate (PP) pathway for glucose catabolism...

  4. Unraveling the complexity of flux regulation: A new method demonstrated for nutrient starvation in Saccharomyces cerevisiae

    OpenAIRE

    Rossell, S.; Weijden, C. C.; Lindenbergh, P. A.; Tuijl, J. H.; Francke, C.; Bakker, B. M.; Westerhoff, H. V.

    2006-01-01

    An important question is to what extent metabolic fluxes are regulated by gene expression or by metabolic regulation. There are two distinct aspects to this question: (i) the local regulation of the fluxes through the individual steps in the pathway and (ii) the influence of such local regulation on the pathway’s flux. We developed regulation analysis so as to address the former aspect for all steps in a pathway. We demonstrate the method for the issue of how Saccharomyces cerevisiae regula...

  5. Role of Doa1 in the Saccharomyces cerevisiae DNA Damage Response†

    OpenAIRE

    Lis, Ewa T.; Romesberg, Floyd E.

    2006-01-01

    The cellular response to DNA damage requires not only direct repair of the damage but also changes in the DNA replication machinery, chromatin, and transcription that facilitate survival. Here, we describe Saccharomyces cerevisiae Doa1, which helps to control the damage response by channeling ubiquitin from the proteosomal degradation pathway into pathways that mediate altered DNA replication and chromatin modification. DOA1 interacts with genes involved in PCNA ubiquitination, including RAD6...

  6. Genetic effects of methyl benzimidazole-2-yl-carbamate on Saccharomyces cerevisiae.

    OpenAIRE

    Wood, J. S.

    1982-01-01

    The genetic effects of the mitotic inhibitor methyl benzimidazole-2-yl-carbamate (MBC) have been studied in Saccharomyces cerevisiae. MBC had little or no effect on the frequency of mutation. In some experiments MBC caused an increase in the frequency of mitotic recombination; however, this effect was small and not reproducible. The primary genetic effect of MBC was to induce mitotic chromosome loss at a high frequency. Chromosome loss occurred at equal frequencies for all chromosomes tested ...

  7. Internuclear transfer of genetic information in kar1-1/KAR1 heterokaryons in Saccharomyces cerevisiae.

    OpenAIRE

    Dutcher, S K

    1981-01-01

    Heterokaryons of Saccharomyces cerevisiae have been constructed utilizing the kar1-1 mutation, which prevents nuclear fusion during conjugation (J. Conde and G. Fink, Proc. Natl. Acad. Sci. U.S.A. 73:3651-3655, 1976). Each heterokaryon contained two haploid nuclei that were marked on several chromosomes. They segregated haploid progeny (cytoductants), most of which have the nuclear genotype of one or the other of the heterokaryon parents, but they occasionally segregated progeny having a reco...

  8. pH-Dependent Uptake of Fumaric Acid in Saccharomyces cerevisiae under Anaerobic Conditions

    OpenAIRE

    Jamalzadeh, E.; Verheijen, P. J.; Heijnen, J.; Gulik, W. M.

    2012-01-01

    Microbial production of C4 dicarboxylic acids from renewable resources has gained renewed interest. The yeast Saccharomyces cerevisiae is known as a robust microorganism and is able to grow at low pH, which makes it a suitable candidate for biological production of organic acids. However, a successful metabolic engineering approach for overproduction of organic acids requires an incorporation of a proper exporter to increase the productivity. Moreover, low-pH fermentations, which are desirabl...

  9. Members of the Hsp70 family of proteins in the cell wall of Saccharomyces cerevisiae.

    OpenAIRE

    Lo?pez-ribot, J. L.; Chaffin, W. L.

    1996-01-01

    Western blot (immunoblot) analysis of cell wall and cytosolic extracts obtained from parental and ssa1 and ssa2 single- and double-mutant strains of Saccharomyces cerevisiae showed that the heat shock protein 70 (Hsp70) products of these genes, previously thought to be restricted to the cell interior, are also present in the cell wall. A cell wall location was further confirmed by indirect immunofluorescence with intact cells and biotinylation of extracellular Hsp70. Hsp70s have been implicat...

  10. Analyse Systémique de la Modulation de la bascule respiro-fermentaire chez Saccharomyces cerevisiae

    OpenAIRE

    Feria Gervasio, David

    2008-01-01

    L’objective de notre travail est d’étudier la transition métabolique respiro-fermentaire chez Saccharomyces cerevisiae, plus spécifiquement d’évaluer l’importance du transport du carbone issu du métabolisme de l’acétyl-coenzymeA vers la mitochondrie. Pour ce faire, une approche originale du génie microbiologique a été mise en place basée sur l’utilisation de chémostat sur substrats mixtes. Cela consiste à introduire au cours d’un chémostat oxydatif sous ...

  11. Ethanol Production from Sago Waste Using Saccharomyces cerevisiae Vits-M1

    OpenAIRE

    Subashini, D.; Suthindhiran, J. Ejilane A. Radha M. A. Jayasri And K.

    2011-01-01

    The present study deals with the biotechnological production of ethanol from sago waste materials. As petroleum has become depleted, renewable energy production has started to gain attention all over the world, including the production of ethanol from sago wastes. In our research we have standardized the production of ethanol from sago wastes using Saccharomyces cerevisiae strain isolated from molasses. The production of ethanol was carried out by means of simultaneous saccharification with a...

  12. Molecular mechanisms of pyrimidine dimer excision in Saccharomyces cerevisiae: excision of dimers in cell extracts

    International Nuclear Information System (INIS)

    Cell-free extracts prepared from rad1-19, rad2-2, rad3-1, rad4-3, rad7-1, rad10-1, rad14-1, rad16-1, and cycl-1 (rad7) mutants of Saccharomyces cerevisiae all catalyze the preferential excision of thymine-containing pyrimidine dimers from ultraviolet-irradiated DNA specifically incised with M. luteus ultraviolet deoxyribonucleic acid incising activity

  13. Synchronization of Cell Cycle of Saccharomyces cerevisiae by Using a Cell Chip Platform

    OpenAIRE

    Hur, Jae Young; Park, Min Cheol; Suh, Kahp-yang; Park, Sang-hyun

    2011-01-01

    Cell synchrony is a critical requirement for the study of eukaryotic cells. Although several chemical and genetic methods of cell cycle synchronization are currently available, they have certain limitations, such as unnecessary perturbations to cells. We developed a novel cell cycle synchronization method that is based on a cell chip platform. The budding yeast, Saccharomyces cerevisiae, is a simple but useful model system to study cell biology and shares many similar features with higher euk...

  14. Fructo-Oligosaccharide Synthesis by Mutant Versions of Saccharomyces cerevisiae Invertase ?†

    OpenAIRE

    Lafraya, A?lvaro; Sanz-aparicio, Julia; Polaina, Julio; Mari?n-navarro, Julia

    2011-01-01

    Efficient enzymatic synthesis of tailor-made prebiotic fructo-oligosaccharides (FOS) used in functional food formulation is a relevant biotechnological objective. We have engineered the Saccharomyces cerevisiae invertase (Suc2) to improve its transferase activity and to identify the enzymatic determinants for product specificity. Amino acid replacement (W19Y, N21S, N24S) within a conserved motif (?-fructosidase) specifically increased the synthesis of 6-kestose up to 10-fold. Mutants with lo...

  15. Isolation and characterization of the RNA2, RNA3, and RNA11 genes of Saccharomyces cerevisiae.

    OpenAIRE

    Last, R. L.; Stavenhagen, J. B.; Woolford, J. L.

    1984-01-01

    Temperature-sensitive mutations in the genes RNA2 through RNA11 cause accumulation of intervening sequence containing precursor mRNAs in Saccharomyces cerevisiae. Three different plasmids have been isolated which complement both the temperature-sensitive lethality and precursor mRNA accumulation when introduced into rna2, rna3, and rna11 mutant strains. The yeast sequences on these plasmids have been shown by Southern transfer hybridization and genetic mapping to be derived from the RNA2, RNA...

  16. Allantoin transport in Saccharomyces cerevisiae is regulated by two induction systems.

    OpenAIRE

    Cooper, T. G.; Chisholm, V. T.; Cho, H. J.; Yoo, H. S.

    1987-01-01

    We show that the allantoin transport system of Saccharomyces cerevisiae responds to two induction systems, one mediated by allophanate or its analog oxalurate and the other mediated by allantoin or its analog hydantoin acetate. The effects of the two inducers were additive in strain M85. Like other allantoin pathway genes, oxalurate-mediated induction of allantoin transport required a functional DAL81 gene product. Hydantoin acetate-mediated induction of the system, on the other hand, occurre...

  17. Isolation and characterization of mutants that produce the allantoin-degrading enzymes constitutively in Saccharomyces cerevisiae.

    OpenAIRE

    Chisholm, G.; Cooper, T. G.

    1982-01-01

    Degradation of allantoin, allantoate, or urea by Saccharomyces cerevisiae requires the participation of four enzymes and four transport systems. Production of the four enzymes and one of the active transport systems is inducible; allophanate, the last intermediate of the pathway, functions as the inducer. The involvement of allophanate in the expression of five distinct genes suggested that they might be regulated by a common element. This suggestion is now supported by the isolation of a new...

  18. Location of the Genes That Control Induction of the Allantoin-Degrading Enzymes in SACCHAROMYCES CEREVISIAE

    OpenAIRE

    Turoscy, Vanessa; Chisholm, George; Cooper, Terrance G.

    1984-01-01

    In an effort to understand the regulation of allantoin degradation in Saccharomyces cerevisiae, we isolated two classes of mutants, each defective in the induction process associated with production of the pathway enzymes. Mutation at one locus (DAL80) results in constitutive expression of the genes involved in allantoin catabolism. Mutation at the second locus (DAL-81) results in the loss of ability to induce these enzymes. This report describes genetic data indicating that the DAL80 and DA...

  19. Ethanol production using Saccharomyces cerevisiae cells immobilised on corn stem ground tissue

    OpenAIRE

    Vu?urovi? Vesna M.; Razmovski Radojka N.; Popov Stevan D.

    2009-01-01

    Cell immobilisation in alcoholic fermentation has been extensively studied during the past few decades because of its technical and economical advantages over those of free cell systems. A biocatalyst was prepared by immobilising a commercial Saccharomyces cerevisiae strain (baker yeast) on corn stem ground tissue for use in alcoholic fermentation. For this purpose, the yeast cells were submitted to the batch tests 'in situ' adsorption onto pieces of the corn stem ground tissue. Cells immobil...

  20. Diversion of Flux toward Sesquiterpene Production in Saccharomyces cerevisiae by Fusion of Host and Heterologous Enzymes? †

    OpenAIRE

    Albertsen, Line; Chen, Yun; Bach, Lars S.; Rattleff, Stig; Maury, Jerome; Brix, Susanne; Nielsen, Jens; Mortensen, Uffe H.

    2010-01-01

    The ability to transfer metabolic pathways from the natural producer organisms to the well-characterized cell factory Saccharomyces cerevisiae is well documented. However, as many secondary metabolites are produced by collaborating enzymes assembled in complexes, metabolite production in yeast may be limited by the inability of the heterologous enzymes to collaborate with the native yeast enzymes. This may cause loss of intermediates by diffusion or degradation or due to conversion of the int...

  1. Daughter cells of Saccharomyces cerevisiae from old mothers display a reduced life span

    OpenAIRE

    1994-01-01

    The yeast Saccharomyces cerevisiae typically divides asymmetrically to give a large mother cell and a smaller daughter cell. As mother cells become old, they enlarge and produce daughter cells that are larger than daughters derived from young mother cells. We found that occasional daughter cells were indistinguishable in size from their mothers, giving rise to a symmetric division. The frequency of symmetric divisions became greater as mother cells aged and reached a maximum occurrence of 30%...

  2. amdSYM, a new dominant recyclable marker cassette for Saccharomyces cerevisiae :

    OpenAIRE

    Solis-escalante, D.; Kuijpers, N. G. A.; Bongaerts, N.; Bolat, I.; Bosman, L.; Pronk, J. T.; Daran, J. M.; Daran-lapujade, P. A. S.

    2012-01-01

    Despite the large collection of selectable marker genes available for Saccharomyces cerevisiae, marker availability can still present a hurdle when dozens of genetic manipulations are required. Recyclable markers, counterselectable cassettes that can be removed from the targeted genome after use, are therefore valuable assets in ambitious metabolic engineering programs. In the present work, the new recyclable dominant marker cassette amdSYM, formed by the Ashbya gossypii TEF2 promoter and ter...

  3. Influence of Indigenous Saccharomyces cerevisiae Strains on Higher Alcohol Content in Malvazija Istarska Wines

    OpenAIRE

    Mario Staver; Stanka Herjavec; Sandi Orli?; Sulejman Redžepovi?; Karin Kova?evi?-Gani?; Ana Jeromel; ?ordano Peršuri?

    2005-01-01

    Istria is one of the most remarkable wine growing regions in Croatia. The main white grape variety is Malvasia from Istria (Malvazija istarska). Commercial yeast inocula, generally Saccharomyces cerevisiae, are widely used as starters today, however it might be preferable to use selected indigenous strains, which may have optimum abilities and may be better adapted to ferment the must of each area. Production and final concentration of higher alcohols is one of the strains most important oeno...

  4. Saccharomyces cerevisiae Srs2 DNA Helicase Selectively Blocks Expansions of Trinucleotide Repeats

    OpenAIRE

    Bhattacharyya, Saumitri; Lahue, Robert S

    2004-01-01

    Trinucleotide repeats (TNRs) undergo frequent mutations in families afflicted with certain neurodegenerative 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, CAG, and CGG repeats. The expansion phenotype was specific, as mutation rates at dinu...

  5. Analysis of Mitotic and Meiotic Defects in Saccharomyces Cerevisiae Srs2 DNA Helicase Mutants

    OpenAIRE

    Palladino, F; Klein, H L

    1992-01-01

    The hyper-gene conversion srs2-101 mutation of the SRS2 DNA helicase gene of Saccharomyces cerevisiae has been reported to suppress the UV sensitivity of rad18 mutants. New alleles of SRS2 were recovered using this suppressor phenotype. The alleles have been characterized with respect to suppression of rad18 UV sensitivity, hyperrecombination, reduction of meiotic viability, and definition of the mutational change within the SRS2 gene. Variability in the degree of rad18 suppression and hyperr...

  6. Molecular Characterization of Propolis-Induced Cell Death in Saccharomyces cerevisiae?†

    OpenAIRE

    Castro, Patri?cia Alves; Savoldi, Marcela; Bonatto, Diego; Barros, Ma?rio Henrique; Goldman, Maria Helena S.; Berretta, Andresa A.; Goldman, Gustavo Henrique

    2011-01-01

    Propolis, a natural product of plant resins, is used by the bees to seal holes in their honeycombs and protect the hive entrance. However, propolis has also been used in folk medicine for centuries. Here, we apply the power of Saccharomyces cerevisiae as a model organism for studies of genetics, cell biology, and genomics to determine how propolis affects fungi at the cellular level. Propolis is able to induce an apoptosis cell death response. However, increased exposure to propolis provides ...

  7. Exploiting Spore-Autonomous Fluorescent Protein Expression to Quantify Meiotic Chromosome Behaviors in Saccharomyces cerevisiae

    OpenAIRE

    Thacker, Drew; Lam, Isabel; Knop, Michael; Keeney, Scott

    2011-01-01

    The budding yeast Saccharomyces cerevisiae has proven to be a rich source of information about the mechanisms and regulation of homologous recombination during meiosis. A common technique for studying this process involves microdissecting the four products (ascospores) of a single meiosis and analyzing the configuration of genetic markers in the spores that are viable. Although this type of analysis is powerful, it can be laborious and time-consuming to characterize the large numbers of meios...

  8. Evaluation of the mutagenicity of aminoglycoside antibiotics in Salmonella typhimurium and Saccharomyces cerevisiae.

    Science.gov (United States)

    Koeda, T; Hirano, F

    1979-06-01

    The mutagenicity of aminoglycoside antibiotics (KM, AKM, DKB, RSM, AMK, GM, TOB) has been studied in cells of the bacteria Salmonella typhimurium and in the yeast Saccharomyces cerevisiae. The bacterial strains (Ames') monitor reverse mutation (point mutation) and the yeast strain D5 monitors mitotic crossing-over, mitotic gene conversion and point mutation. None of these antibiotics demonstrated any mutagenic activities in either the bacteria or the yeast. PMID:381273

  9. Interfering with Glycolysis Causes Sir2-Dependent Hyper-Recombination of Saccharomyces cerevisiae Plasmids

    OpenAIRE

    Ralser, Markus; Zeidler, Ute; Lehrach, Hans

    2009-01-01

    Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key metabolic regulator implicated in a variety of cellular processes. It functions as a glycolytic enzyme, a protein kinase, and a metabolic switch under oxidative stress. Its enzymatic inactivation causes a major shift in the primary carbohydrate flux. Furthermore, the protein is implicated in regulating transcription, ER-to-Golgi transport, and apoptosis. We found that Saccharomyces cerevisiae cells null for all GAPDH paralogues (Tdh1, ...

  10. Salt tolerance and methionine biosynthesis in Saccharomyces cerevisiae involve a putative phosphatase gene.

    OpenAIRE

    Gläser, H U; Thomas, D.; Gaxiola, R.; Montrichard, F; Surdin-Kerjan, Y; Serrano, R

    1993-01-01

    The progressive salinization of irrigated land poses a threat to the future of agriculture in arid regions. The identification of crucial metabolic steps in salt tolerance is important for the understanding of stress physiology and may provide the tools for its genetic engineering. In the yeast Saccharomyces cerevisiae we have isolated a gene, HAL2, which upon increase in gene dosage improves growth under NaCl and LiCl stresses. The HAL2 protein is homologous to inositol phosphatases, enzymes...

  11. Transcriptional Response of Saccharomyces cerevisiae to the Plasma Membrane-Perturbing Compound Chitosan

    OpenAIRE

    Zakrzewska, Anna; Boorsma, Andre; Brul, Stanley; Hellingwerf, Klaas J; Klis, Frans M

    2005-01-01

    Chitosan is a plasma membrane-perturbing compound consisting of linear chains of ?-1,4-linked glucosamine residues, which at acidic pHs become positively charged. It is extensively used as an antimicrobial compound, yet its mode of action is still unresolved. Chitosan strongly affected the growth of the yeast Saccharomyces cerevisiae, the food spoilage yeast Zygosaccharomyces bailii, and two human-pathogenic yeasts, Candida albicans and Candida glabrata. Microarray analysis of yeast cells tre...

  12. Suppression of the profilin-deficient phenotype by the RHO2 signaling pathway in Saccharomyces cerevisiae.

    OpenAIRE

    Marcoux, N; Cloutier, S.; Zakrzewska, E; Charest, P M; Bourbonnais, Y; Pallotta, D

    2000-01-01

    Profilin plays an important role in actin organization in all eukaryotic cells through mechanisms that are still poorly understood. We had previously shown that Mid2p, a transmembrane protein and a potential cell wall sensor, is an effective multicopy suppressor of the profilin-deficient phenotype in Saccharomyces cerevisiae. To better understand the role of Mid2p in the organization of the actin cytoskeleton, we isolated five additional multicopy suppressors of pfy1Delta cells that are Rom1p...

  13. The Sum1/Ndt80 Transcriptional Switch and Commitment to Meiosis in Saccharomyces cerevisiae

    OpenAIRE

    Winter, Edward

    2012-01-01

    Summary: Cells encounter numerous signals during the development of an organism that induce division, differentiation, and apoptosis. These signals need to be present for defined intervals in order to induce stable changes in the cellular phenotype. The point after which an inducing signal is no longer needed for completion of a differentiation program can be termed the “commitment point.” Meiotic development in the yeast Saccharomyces cerevisiae (sporulation) provides a model system to s...

  14. NDT80, a meiosis-specific gene required for exit from pachytene in Saccharomyces cerevisiae.

    OpenAIRE

    Xu, L.; Ajimura, M; Padmore, R; Klein, C.; Kleckner, N.

    1995-01-01

    We describe the identification of a new meiosis-specific gene of Saccharomyces cerevisiae, NDT80. The ndt80 null and point mutants arrest at the pachytene stage of meiosis, with homologs connected by full-length synaptonemal complexes and spindle pole bodies duplicated but unseparated. Meiotic recombination in an ndt80 delta mutant is relatively normal, although commitment to heteroallelic recombination is elevated two- to threefold and crossing over is decreased twofold compared with those o...

  15. Malic acid production by Saccharomyces cerevisiae: engineering of pyruvate carbosylation, oxaloacetate reduction and malate export

    OpenAIRE

    Zelle, R. M.; Hulster, E.; Winden, W. A.; Waard, P.; Dijkema, C.; Winkler, A. A.; Geertman, J. M. A.

    2008-01-01

    Malic acid is a potential biomass-derivable "building block" for chemical synthesis. Since wild-type Saccharomyces cerevisiae strains produce only low levels of malate, metabolic engineering is required to achieve efficient malate production with this yeast. A promising pathway for malate production from glucose proceeds via carboxylation of pyruvate, followed by reduction of oxaloacetate to malate. This redox- and ATP-neutral, CO2-fixing pathway has a theoretical maximum yield of 2 mol malat...

  16. Growth Temperature Exerts Differential Physiological and Transcriptional Responses in Laboratory and Wine Strains of Saccharomyces cerevisiae? †

    OpenAIRE

    Pizarro, Francisco J.; Jewett, Michael C.; Nielsen, Jens; Agosin, Eduardo

    2008-01-01

    Laboratory strains of Saccharomyces cerevisiae have been widely used as a model for studying eukaryotic cells and mapping the molecular mechanisms of many different human diseases. Industrial wine yeasts, on the other hand, have been selected on the basis of their adaptation to stringent environmental conditions and the organoleptic properties that they confer to wine. Here, we used a two-factor design to study the responses of a standard laboratory strain, CEN.PK113-7D, and an industrial win...

  17. Malic Acid Production by Saccharomyces cerevisiae: Engineering of Pyruvate Carboxylation, Oxaloacetate Reduction, and Malate Export? †

    OpenAIRE

    Zelle, R. M.; Hulster, E.; Winden, W. A.; Waard, P.; Dijkema, C.; Winkler, A. A.; Geertman, J. M.; Dijken, J. P.; Pronk, J. T.; Maris, A. J. A.

    2008-01-01

    Malic acid is a potential biomass-derivable "building block" for chemical synthesis. Since wild-type Saccharomyces cerevisiae strains produce only low levels of malate, metabolic engineering is required to achieve efficient malate production with this yeast. A promising pathway for malate production from glucose proceeds via carboxylation of pyruvate, followed by reduction of oxaloacetate to malate. This redox- and ATP-neutral, CO(2)-fixing pathway has a theoretical maximum yield of 2 mol mal...

  18. Transcriptional Response to Deletion of the Phosphatidylserine Decarboxylase Psd1p in the Yeast Saccharomyces cerevisiae

    OpenAIRE

    Gsell, Martina; Mascher, Gerald; Schuiki, Irmgard; Ploier, Birgit; Hrastnik, Claudia; Daum, Gu?nther

    2013-01-01

    In the yeast, Saccharomyces cerevisiae, the synthesis of the essential phospholipid phosphatidylethanolamine (PE) is accomplished by a network of reactions which comprises four different pathways. The enzyme contributing most to PE formation is the mitochondrial phosphatidylserine decarboxylase 1 (Psd1p) which catalyzes conversion of phosphatidylserine (PS) to PE. To study the genome wide effect of an unbalanced cellular and mitochondrial PE level and in particular the contribution of Psd1p t...

  19. Saccharomyces cerevisiae oxidative response evaluation by cyclic voltammetry and gas chromatography?mass spectrometry

    OpenAIRE

    Castro, Cristiana C.; Gunning, Caitriona; Oliveira, Carla M.; Couto, Jose? A.; Teixeira, J. A.; Martins, Rui C.; Ferreira, Anto?nio C. Silva

    2012-01-01

    This study is focused on the evaluation of the impact of Saccharomyces cerevisiae metabolism in the profile of compounds with antioxidant capacity in a synthetic wine during fermentation. A bioanalytical pipeline, which allows for biological systems fingerprinting and sample classification by combining electrochemical features with biochemical background, is proposed. To achieve this objective, alcoholic fermentations of a minimal medium supplemented with phenolic acids were evaluated daily d...

  20. Involvement of the Saccharomyces cerevisiae Hydrolase Ldh1p in Lipid Homeostasis?

    OpenAIRE

    Debelyy, Mykhaylo O.; Thoms, Sven; Connerth, Melanie; Daum, Gu?nther; Erdmann, Ralf

    2011-01-01

    Here, we report the functional characterization of the newly identified lipid droplet hydrolase Ldh1p. Recombinant Ldh1p exhibits esterase and triacylglycerol lipase activities. Mutation of the serine in the hydrolase/lipase motif GXSXG completely abolished esterase activity. Ldh1p is required for the maintenance of a steady-state level of the nonpolar and polar lipids of lipid droplets. A characteristic feature of the Saccharomyces cerevisiae ?ldh1 strain is the appearance of giant lipid dr...

  1. Genetic Analysis of Azole Resistance by Transposon Mutagenesis in Saccharomyces cerevisiae

    OpenAIRE

    Kontoyiannis, D. P.

    1999-01-01

    The increasing resistance of Candida species to fluconazole is cause for concern. To determine the molecular mechanisms involved in resistance to fluconazole, I used a scheme of transposon mutagenesis in Saccharomyces cerevisiae, a genetically tractable yeast that is closely related to Candida albicans. This technique, which permits the generation and analysis of multiple random Tn3::LEU2::lacZ fusions, can be used as a disruption mutagen (N. B. Burns et al., Genes Dev. 8:1087–1105, 1994). By...

  2. Isolation and characterization of vanadate-resistant mutants of Saccharomyces cerevisiae.

    OpenAIRE

    Willsky, G. R.; Leung, J. O.; Offermann, P. V.; Plotnick, E. K.; Dosch, S. F.

    1985-01-01

    Cellular vanadium metabolism was studied in Saccharomyces cerevisiae by isolating and characterizing vanadate [VO4(3-), V(V)]-resistant mutants. Vanadate growth inhibition was reversed by the removal of the vanadate from the medium, and vanadate resistance was found to be a recessive trait. Vanadate-resistant mutants isolated from glucose-grown cells were divided into five complementation classes containing more than one mutant. Among the vanadate-resistant mutants isolated in maltose medium,...

  3. Alterations in translatable ribonucleic acid after heat shock of Saccharomyces cerevisiae.

    OpenAIRE

    Mcalister, L.; Finkelstein, D. B.

    1980-01-01

    Changes in populations of translatable messenger ribonucleic acids (mRNA's) after heat shock of Saccharomyces cerevisiae were examined and found to correlate very closely with transient alterations in patterns of in vivo protein synthesis. Initial changes included an increase in translatable species coding for polypeptides synthesized during heat shock; this increase was found to be dependent on transcription but did not require ongoing protein synthesis. A decrease was observed in the level ...

  4. Heat shock proteins affect RNA processing during the heat shock response of Saccharomyces cerevisiae.

    OpenAIRE

    Yost, H. J.; Lindquist, S.

    1991-01-01

    In the yeast Saccharomyces cerevisiae, the splicing of mRNA precursors is disrupted by a severe heat shock. Mild heat treatments prior to severe heat shock protect splicing from disruption, as was previously reported for Drosophila melanogaster. In contrast to D. melanogaster, protein synthesis during the pretreatment is not required to protect splicing in yeast cells. However, protein synthesis is required for the rapid recovery of splicing once it has been disrupted by a sudden severe heat ...

  5. Evidence for cooperation between cells during sporulation of the yeast Saccharomyces cerevisiae.

    OpenAIRE

    Jakubowski, H.; Goldman, E.

    1988-01-01

    Diploid Saccharomyces cerevisiae cells heterozygous for the mating type locus (MATa/MAT alpha) undergo meiosis and sporulation when starved for nitrogen in the presence of a poor carbon source such as potassium acetate. Diploid yeast adenine auxotrophs sporulated well at high cell density (10(7) cells per ml) under these conditions but failed to differentiate at low cell density (10(5) cells per ml). The conditional sporulation-deficient phenotype of adenine auxotrophs could be complemented b...

  6. The daughters of Saccharomyces cerevisiaeRAS2val19 mutant are born old.

    Czech Academy of Sciences Publication Activity Database

    Pichová, Alena; Sigler, Karel

    Bratislava : SAS, 2007, s. 80-80. ISSN 1336-4839. [Annual Conference on Yeasts /35./. Smolenice (SK), 16.05.2007-18.05.2007] R&D Projects: GA ?R GA301/03/0289; GA ?R GA301/07/0339; GA MŠk 1M0570 Institutional research plan: CEZ:AV0Z50200510 Keywords : saccharomyces cerevisiae Subject RIV: EE - Microbiology, Virology

  7. Population Size Drives Industrial Saccharomyces cerevisiae Alcoholic Fermentation and Is under Genetic Control?†‡

    OpenAIRE

    Albertin, Warren; Marullo, Philippe; Aigle, Michel; Dillmann, Christine; Vienne, Dominique; Bely, Marina; Sicard, Delphine

    2011-01-01

    Alcoholic fermentation (AF) conducted by Saccharomyces cerevisiae has been exploited for millennia in three important human food processes: beer and wine production and bread leavening. Most of the efforts to understand and improve AF have been made separately for each process, with strains that are supposedly well adapted. In this work, we propose a first comparison of yeast AFs in three synthetic media mimicking the dough/wort/grape must found in baking, brewing, and wine making. The fermen...

  8. Engineering of Polyploid Saccharomyces cerevisiae for Secretion of Large Amounts of Fungal Glucoamylase

    OpenAIRE

    Ekino, Keisuke; Hayashi, Hiroyuki; Moriyama,Masahiro; Matsuda, Minoru; Goto, Masatoshi; Yoshino, Sadazo; Furukawa, Kensuke

    2002-01-01

    We engineered Saccharomyces cerevisiae cells that produce large amounts of fungal glucoamylase (GAI) from Aspergillus awamori var. kawachi. To do this, we used the ?-sequence-mediated integration vector system and the heat-induced endomitotic diploidization method. ?-Sequence-mediated integration is known to occur mainly in a particular chromosome, and the copy number of the integration is variable. In order to construct transformants carrying the GAI gene on several chromosomes, haploid ce...

  9. Transcriptional control of the sporulation-specific glucoamylase gene in the yeast Saccharomyces cerevisiae.

    OpenAIRE

    Yamashita, I; Fukui, S

    1985-01-01

    In the yeast Saccharomyces cerevisiae, glucoamylase activity appears specifically in sporulating cells heterozygous for the mating-type locus (MAT). We identified a sporulation-specific glucoamylase gene (SGA) and show that expression of SGA is positively regulated by the mating-type genes, both MATa1 and MAT alpha 2. Northern blot analysis revealed that control of SGA is exerted at the level of RNA production. Expression of SGA or the consequent degradation of glycogen to glucose in cells is...

  10. Severe Adenine Starvation Activates Ty1 Transcription and Retrotransposition in Saccharomyces cerevisiae

    OpenAIRE

    Todeschini, Anne-laure; Morillon, Antonin; Springer, Mathias; Lesage, Pascale

    2005-01-01

    Ty1 retrotransposons of the yeast Saccharomyces cerevisiae are activated by different kinds of stress. Here we show that Ty1 transcription is stimulated under severe adenine starvation conditions. The Bas1 transcriptional activator, responsible for the induction of genes of the de novo AMP biosynthesis pathway (ADE) in the absence of adenine, is not involved in this response. Activation occurs mainly on Ty1 elements, whose expression is normally repressed by chromatin and is suppressed in a h...

  11. Global Analyses of Sumoylated Proteins in Saccharomyces cerevisiae: Induction of Protein Sumoylation by Cellular Stresses*

    OpenAIRE

    Zhou, Weidong; Ryan, Jennifer J.; Zhou, Huilin

    2004-01-01

    We have undertaken a global analysis of sumoylated proteins in Saccharomyces cerevisiae by tandem mass spectrometry. Exposure of cells to oxidative and ethanol stresses caused large increases in protein sumoylation. A large number of new sumoylated proteins were identified in untreated, hydrogen peroxide-treated, and ethanol-treated cells. These proteins are known to be involved in diverse cellular processes, including gene transcription, protein translation, DNA replication, chromosome segre...

  12. Gcn4 Is Required for the Response to Peroxide Stress in the Yeast Saccharomyces cerevisiae

    OpenAIRE

    Mascarenhas, Claire; Edwards-ingram, Laura C.; Zeef, Leo; Shenton, Daniel; Ashe, Mark P.; Grant, Chris M.

    2008-01-01

    An oxidative stress occurs when reactive oxygen species overwhelm the cellular antioxidant defenses. We have examined the regulation of protein synthesis in Saccharomyces cerevisiae in response to oxidative stress induced by exposure to hydroperoxides (hydrogen peroxide, and cumene hydroperoxide), a thiol oxidant (diamide), and a heavy metal (cadmium). Examination of translational activity indicates that these oxidants inhibit translation at the initiation and postinitiation phases. Inhibitio...

  13. Utilização da levedura desidratada (Saccharomyces cerevisiae) para leitões na fase inicial

    OpenAIRE

    Araújo Lúcio Francelino; Junqueira Otto Mack; Lopes Eurípedes Laurindo; Araújo Cristiane Soares da Silva; Ortolan Josiane Hernandes; Laurentiz Antônio Carlos de

    2006-01-01

    Foi conduzido um experimento com o objetivo de avaliar o efeito da adição de diferentes níveis de levedura (Saccharomyces cerevisiae) desidratada na ração sobre o desempenho e a morfologia intestinal de leitões na fase inicial. Foram utilizados 280 leitões (fêmeas e machos castrados) de uma linha genética comercial de suínos, desmamados com 21 dias de idade e distribuídos em 20 baias, de acordo com o delineamento em blocos ao acaso, com 5 repetições e 4 tratamentos experimentais ...

  14. Robust industrial Saccharomyces cerevisiae strains for very high gravity bio-ethanol fermentations

    OpenAIRE

    Pereira, Francisco B; Guimarães, Pedro M. R.; Teixeira, J. A.; Domingues, Lucília

    2011-01-01

    The application and physiological background of two industrial Saccharomyces cerevisiae strains, isolated from harsh industrial environments, were studied in Very High Gravity (VHG) bio-ethanol fermentations. VHG laboratory fermentations, mimicking industrially relevant conditions, were performed with PE-2 and CA1185 industrial strains and the CEN.PK113-7D laboratory strain. The industrial isolates produced remarkable high ethanol titres (>19%, v/v) and accumulated an increased content of ste...

  15. Heterologous Expression of an Entamoeba histolytica Chitin Synthase in Saccharomyces cerevisiae

    OpenAIRE

    Van Dellen, Katrina L.; Bulik, Dorota A.; Specht, Charles A; Robbins, Phillips W.; Samuelson, John C.

    2006-01-01

    Chitin in the cyst wall of Entamoeba histolytica is made by two chitin synthases (Chs), one of which is unique (EhCHS-1) and one of which resembles those of insects and nematodes (EhCHS-2). EhCHS-1 is deposited chitin in the lateral wall of transformed Saccharomyces cerevisiae Chs mutants, independent of accessory proteins (Chs4p to Chs7p) required by yeast Chs3p.

  16. Chitin synthase 2 is essential for septum formation and cell division in Saccharomyces cerevisiae.

    OpenAIRE

    Silverman, S J; Sburlati, A; Slater, M L; Cabib, E.

    1988-01-01

    Previous work led to the puzzling conclusion that chitin synthase 1, the major chitin synthase activity in Saccharomyces cerevisiae, is not required for synthesis of the chitinous primary septum. The mechanism of in vivo synthesis of chitin has now been clarified by cloning the structural gene for the newly found chitin synthase 2, a relatively minor activity in yeast. Disruption of the chitin synthase 2 gene results in the loss of well-defined septa and in growth arrest, establishing that th...

  17. Localization of chitin synthetase in cell-free homogenates of Saccharomyces cerevisiae: chitosomes and plasma membrane.

    OpenAIRE

    Leal-Morales, C A; Bracker, C. E.; Bartnicki-Garcia, S

    1988-01-01

    We describe an improved method for fractionating cell-free extracts of Saccharomyces cerevisiae to separate its membranous components by a combination of isopycnic and velocity sedimentations. These procedures were used to examine the subcellular distribution of chitin synthetase (chitin-UDP acetylglucosaminyltransferase; EC 2.4.1.16) in homogenates from exponentially growing walled cells of a wild-type strain of yeast. Chitin synthetase (Chs1) activity was mainly found in two distinct vesicl...

  18. FERMENTATION, MEDIA OPTIMIZATION STUDIES FOR COENZYME Q10 PRODUCTION BY Saccharomyces cerevisiae

    OpenAIRE

    Narendra Kumar S; Puspha Agrawal; Sujata A S; Bhavana B K

    2012-01-01

    To establish the fermentation process for CoQ10 production by using Saccharomyces cerevisiae with subsequent bioprocess media optimization studies. Coenzyme Q10 (CoQ10) is a vitamin-like nutrient that plays a vital role in cellular energy production. Coenzyme Q10, apart from playing an important role in electron transfer for ATP synthesis, also acts as antioxidant. Therefore it is preferred as a potential therapeutic supplement to many critical diseases besides being used as dietary supplemen...

  19. ENHANCEMENT OF SACCHAROMYCES CEREVISIAE GLUTATHIONE AND MICRONUTRIENTS CONTENT FOR NUTRACEUTICAL APPLICATIONS.

    OpenAIRE

    Musatti, Alida

    2012-01-01

    This Ph.D. research concerns nutritional and microbiological aspects, and the final aim of this project is the development of innovative food supplements formulations containing Saccharomyces cerevisiae, enriched, by biotechnological processes, with micronutrients and antioxidant molecules. This idea was supported by the fact that even if in the developed world a clinical deficiency of micronutrients is uncommon, a suboptimal intake of certain micronutrients has been linked with an increa...

  20. Functional complementation in Saccharomyces cerevisiae under control of the natural yeast promoter

    Scientific Electronic Library Online (English)

    Rob, Lavigne; Guido, Volckaert.

    2007-04-15

    Full Text Available A novel approach for functional complementation of foreign genes in Saccharomyces cerevisiae is presented. This approach is based on the use of the widely available cognate gene plasmids (e.g. pRS416) of the European Functional Analysis Network (EUROFAN). The functional complementation of the human [...] homolog of YOR159c (SME1 gene) shown here is the first demonstration of complementation using the original yeast promoter, theoretically offering a more natural regulation of protein expression

  1. Biochemical studies of the Saccharomyces cerevisiae Mph1 helicase on junction-containing DNA structures

    OpenAIRE

    Kang, Young-Hoon; Munashingha, Palinda Ruvan; Lee, Chul-Hwan; Nguyen, Tuan Anh; Seo, Yeon-Soo

    2011-01-01

    Saccharomyces cerevisiae Mph1 is a 3–5? DNA helicase, required for the maintenance of genome integrity. In order to understand the ATPase/helicase role of Mph1 in genome stability, we characterized its helicase activity with a variety of DNA substrates, focusing on its action on junction structures containing three or four DNA strands. Consistent with its 3? to 5? directionality, Mph1 displaced 3?-flap substrates in double-fixed or equilibrating flap substrates. Surprisingly, Mph1 d...

  2. In Vivo Analysis of the Mechanisms for Oxidation of Cytosolic NADH by Saccharomyces cerevisiae Mitochondria

    OpenAIRE

    Overkamp, K.M.; Bakker, B.M.; Kotter, P.; Tuijl, A., van; de Vries, S; Dijken, J.P. van; Pronk, J T

    2000-01-01

    During respiratory glucose dissimilation, eukaryotes produce cytosolic NADH via glycolysis. This NADH has to be reoxidized outside the mitochondria, because the mitochondrial inner membrane is impermeable to NADH. In Saccharomyces cerevisiae, this may involve external NADH dehydrogenases (Nde1p or Nde2p) and/or a glycerol-3-phosphate shuttle consisting of soluble (Gpd1p or Gpd2p) and membrane-bound (Gut2p) glycerol-3-phosphate dehydrogenases. This study addresses the physiological relevance o...

  3. Characterization of vanadate-dependent NADH oxidation stimulated by Saccharomyces cerevisiae plasma membranes.

    OpenAIRE

    Minasi, L A; Willsky, G R

    1991-01-01

    Plasma membrane-stimulated vanadate-dependent NADH oxidation has been characterized in Saccharomyces cerevisiae. This activity is specific for vanadate, because molybdate, a similar metal oxide, did not substitute for vanadate in the reaction. Vanadate-dependent plasma membrane-stimulated NADH oxidation activity was dependent on the concentrations of vanadate, NADH, and NADPH and required functional plasma membranes; no stimulation occurred in the presence of boiled membranes or bovine serum ...

  4. Flocculation onset in Saccharomyces cerevisiae: effect of ethanol, heat and osmotic stress

    OpenAIRE

    Claro, F. B.; Rijsbrack, K.; Soares, Eduardo V.

    2007-01-01

    Aims: To examine the effect of different stress conditions on the onset of flocculation in an ale-brewing strain, Saccharomyces cerevisiae NCYC 1195. Methods and Results: Flocculation was evaluated using the method of Soares, E.V. and Vroman, A. [Journal of Applied Microbiology (2003) 95, 325]; plasma membrane integrity was accessed using propidium iodide and the staining of the yeast cell wall was performed using calcofluor white M2R. Cells in exponential phase of growth we...

  5. Effects of particulate materials and osmoprotectants on very-high-gravity ethanolic fermentation by Saccharomyces cerevisiae.

    OpenAIRE

    Thomas, K. C.; Hynes, S. H.; Ingledew, W. M.

    1994-01-01

    The effects of osmoprotectants (such as glycine betaine and proline) and particulate materials on the fermentation of very high concentrations of glucose by the brewing strain Saccharomyces cerevisiae (uvarum) NCYC 1324 were studied. The yeast growing at 20 degrees C consumed only 15 g of the sugar per 100 ml from a minimal medium which initially contained 35% (wt/vol) glucose. Supplementing the medium with a mixture of glycine betaine, glycine, and proline increased the amount of sugar ferme...

  6. Enhancement of Ethanol Fermentation in Saccharomyces cerevisiae Sake Yeast by Disrupting Mitophagy Function

    OpenAIRE

    Shiroma, Shodai; Jayakody, Lahiru Niroshan; Horie, Kenta; Okamoto, Koji; Kitagaki, Hiroshi

    2014-01-01

    Saccharomyces cerevisiae sake yeast strain Kyokai no. 7 has one of the highest fermentation rates among brewery yeasts used worldwide; therefore, it is assumed that it is not possible to enhance its fermentation rate. However, in this study, we found that fermentation by sake yeast can be enhanced by inhibiting mitophagy. We observed mitophagy in wild-type sake yeast during the brewing of Ginjo sake, but not when the mitophagy gene (ATG32) was disrupted. During sake brewing, the maximum rate ...

  7. Production of white wine by Saccharomyces cerevisiae immobilized on grape pomace

    OpenAIRE

    Genisheva, Z.; Macedo, S.; Mussatto, Solange I.; Teixeira, J. A.; Oliveira, J. M.

    2012-01-01

    White wine was produced with Saccharomyces cerevisiae cells immobilized on grape pomace, by natural adsorption. The support, the main solid waste from the wine industry, consisted of the skins, seeds and stems. Immobilization was tested using different media, namely complex culture medium, raw grape must and diluted grape must. Grape pomace was revealed to be an appropriate support for yeast cell immobilization. Moreover, grape must was shown to be the most suitable medium as immobilized c...

  8. Genetic Dissection of Ethanol Tolerance in the Budding Yeast Saccharomyces cerevisiae

    OpenAIRE

    Hu, X. H.; Wang, M. H.; Tan, T.; Li, J. R.; Yang, H.; Leach, L.; Zhang, R. M.; Luo, Z. W.

    2007-01-01

    Uncovering genetic control of variation in ethanol tolerance in natural populations of yeast Saccharomyces cerevisiae is essential for understanding the evolution of fermentation, the dominant lifestyle of the species, and for improving efficiency of selection for strains with high ethanol tolerance, a character of great economic value for the brewing and biofuel industries. To date, as many as 251 genes have been predicted to be involved in influencing this character. Candidacy of these gene...

  9. The relationship between viability and intracellular pH in the yeast Saccharomyces cerevisiae.

    OpenAIRE

    Imai, T.; Ohno, T.

    1995-01-01

    The relationship between viability (cell proliferation activity) and intracellular pH in the yeast Saccharomyces cerevisiae was investigated by using cells that had been deactivated by low-temperature storage, ethanol treatment, or heat treatment. The intracellular pH was measured with a microscopic image processor or a spectrofluorophotometer. At first, the intracellular pH measurements of individual cells were compared with slide culture results by microscopic image processing. A clear corr...

  10. Increased isobutanol production in Saccharomyces cerevisiae by overexpression of genes in valine metabolism

    OpenAIRE

    Karhumaa Kaisa; Kielland-Brandt Morten C; Borodina Irina; Nielsen Kristian F; Chen Xiao

    2011-01-01

    Abstract Background Isobutanol can be a better biofuel than ethanol due to its higher energy density and lower hygroscopicity. Furthermore, the branched-chain structure of isobutanol gives a higher octane number than the isomeric n-butanol. Saccharomyces cerevisiae was chosen as the production host because of its relative tolerance to alcohols, robustness in industrial fermentations, and the possibility for future combination of isobutanol production with fermentation of lignocellulosic mater...

  11. Formation of AAV Single Stranded DNA Genome from a Circular Plasmid in Saccharomyces cerevisiae

    OpenAIRE

    Cervelli, Tiziana; Backovic, Ana; Galli, Alvaro

    2011-01-01

    Adeno-associated virus (AAV)-based vectors are promising tools for targeted transfer in gene therapy studies. Many efforts have been accomplished to improve production and purification methods. We thought to develop a simple eukaryotic system allowing AAV replication which could provide an excellent opportunity for studying AAV biology and, more importantly, for AAV vector production. It has been shown that yeast Saccharomyces cerevisiae is able to replicate and form the capsid of many viruse...

  12. Hybridization and Polyploidization of Saccharomyces cerevisiae Strains by Transformation-Associated Cell Fusion

    OpenAIRE

    Takagi, Atsuko; Harashima, Satoshi; Oshima, Yasuji

    1985-01-01

    Hybrid or polyploid clones of Saccharomyces cerevisiae produced by protoplast fusion were easily isolated by selecting transformants with the plasmid phenotype because the transformation was directly associated with cell fusion. When haploid cells were used as the original strain, the transformants were mostly diploids with a significant fraction of polyploids (triploids or tetraploids). Repeated transformation after curing the plasmid gave rise to clones with higher ploidy, but the frequency...

  13. Construction and Characterization of Isogenic Series of Saccharomyces cerevisiae Polyploid Strains

    OpenAIRE

    Takagi, Atsuko; Harashima, Satoshi; Oshima, Yasuji

    1983-01-01

    Tetraploid cells of Saccharomyces cerevisiae are generated spontaneously in a homothallic MATa/MAT? diploid population at low frequency (approximately 10?6 per cell) through the homozygosity of mating-type alleles by mitotic recombination followed by homothallic switching of the mating-type alleles. To isolate tetraploid clones more effectively, a selection method was developed that used a dye plate containing 40 mg each of eosin Y and amaranth in synthetic nutrient agar per liter. It was ...

  14. Saccharomyces cerevisiae Grx6 and Grx7 are monothiol glutaredoxins associated with the early secretory pathway

    OpenAIRE

    Izquierdo Álvarez, Alicia; Casas Herranz, Celia; Mühlenhoff, Ulrich; Lillig, Christopher Horst; Herrero Perpiñán, Enrique

    2008-01-01

    Saccharomyces cerevisiae Grx6 and Grx7 are two monothiol glutaredoxins whose active-site sequences (CSYS and CPYS, respectively) are reminiscent of the CPYC active-site sequence of classical dithiol glutaredoxins. Both proteins contain an N-terminal transmembrane domain which is responsible for their association to membranes of the early secretory pathway vesicles, facing the luminal side. Thus, Grx6 localizes at the endoplasmic reticulum and Golgi compartments, while Grx7 is mostly at the Go...

  15. Integration of Stress Responses: Modulation of Calcineurin Signaling in Saccharomyces cerevisiae by Protein Kinase A

    OpenAIRE

    Kafadar, Kimberly A.; Cyert, Martha S.

    2004-01-01

    Calcineurin is a Ca2+/calmodulin-dependent protein phosphatase required for Saccharomyces cerevisiae to adapt to a variety of environmental stresses. Once activated, calcineurin dephosphorylates the Zn-finger transcription factor Crz1p/Tcn1p, causing it to accumulate in the nucleus where it activates gene expression. Here we show that cyclic AMP-dependent protein kinase A (PKA) phosphorylates and negatively regulates Crz1p activity by inhibiting its nuclear import. Activation of PKA in vivo d...

  16. Genetic Analysis of Mutants of SACCHAROMYCES CEREVISIAE Resistant to the Herbicide Sulfometuron Methyl

    OpenAIRE

    Falco, S. C.; Dumas, K. S.

    1985-01-01

    Sulfometuron methyl (SM), a potent new sulfonylurea herbicide, inhibits growth of the yeast Saccharomyces cerevisiae on minimal media. Sixty-six spontaneous mutants resistant to SM were isolated. All of the resistance mutations segregate 2:2 in tetrads; 51 of the mutations are dominant, five are semidominant and ten are recessive. The mutations occur in three linkage groups, designated SMR1, smr2 and smr3. Several lines of evidence demonstrate that the SMR1 mutations (47 dominant and four sem...

  17. Metabolic Flux Analysis during the Exponential Growth Phase of Saccharomyces cerevisiae in Wine Fermentations

    OpenAIRE

    Quirós, Manuel; Martínez-Moreno, Rubén; Albiol, Joan; Morales, Pilar; Vázquez-Lima, Felícitas; Barreiro-Vázquez, Antonio; Ferrer, Pau; Gonzalez, Ramon

    2013-01-01

    As a consequence of the increase in global average temperature, grapes with the adequate phenolic and aromatic maturity tend to be overripe by the time of harvest, resulting in increased sugar concentrations and imbalanced C/N ratios in fermenting musts. This fact sets obvious additional hurdles in the challenge of obtaining wines with reduced alcohols levels, a new trend in consumer demands. It would therefore be interesting to understand Saccharomyces cerevisiae physiology during the fermen...

  18. Switch between Life History Strategies Due to Changes in Glycolytic Enzyme Gene Dosage in Saccharomyces cerevisiae? †

    OpenAIRE

    Wang, Shaoxiao; Spor, Aymé; Nidelet, Thibault; Montalent, Pierre; DILLMANN, CHRISTINE; de Vienne, Dominique; Sicard, Delphine

    2010-01-01

    Adaptation is the process whereby a population or species becomes better fitted to its habitat through modifications of various life history traits which can be positively or negatively correlated. The molecular factors underlying these covariations remain to be elucidated. Using Saccharomyces cerevisiae as a model system, we have investigated the effects on life history traits of varying the dosage of genes involved in the transformation of resources into energy. Changing gene dosage for eac...

  19. Integration of Agrobacterium tumefaciens T-DNA in the Saccharomyces cerevisiae genome by illegitimate?recombination

    OpenAIRE

    Bundock, Paul; Hooykaas, Paul?J ?J

    1996-01-01

    Agrobacterium tumefaciens can transfer part of its Ti plasmid, the T-DNA, to plant cells where it integrates into the nuclear genome via illegitimate recombination. Integration of the T-DNA results in small deletions of the plant target DNA, and may lead to truncation of the T-DNA borders and the production of filler DNA. We showed previously that T-DNA can also be transferred from A. tumefaciens to Saccharomyces cerevisiae and integrates into the yeast genome via ...

  20. Mitotic Transmission of Artificial Chromosomes in Cdc Mutants of the Yeast, Saccharomyces Cerevisiae

    OpenAIRE

    Palmer, R. E.; Hogan, E.; Koshland, D

    1990-01-01

    In the yeast, Saccharomyces cerevisiae, cell division cycle (CDC) genes have been identified whose products are required for the execution of different steps in the cell cycle. In this study, the fidelity of transmission of a 14-kb circular minichromosome and a 155-kb linear chromosome fragment was examined in cell divisions where specific CDC products were temporarily inactivated with either inhibitors, or temperature sensitive mutations in the appropriate CDC gene. All of the cdc mutants pr...