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  • 1
    Electronic Resource
    Electronic Resource
    Cloning and sequence analysis of lily and tobacco guanylate kinases (2000)
    Springer
    Molecular biology reports 27 (2000), S. 45-49 
    Details
    ISSN: 1573-4978
    Keywords: Arabidopsis ; guanylate kinase ; lily ; tobacco ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Guanylate kinase is an essential enzyme in the nucleotide biosynthetic pathway, catalyzing the reversible transfer of the terminal phospharyl group of ATP to GMP or dGMP. This enzyme has been well studied from several organisms and many structural and functional details have been characterized. Animal GMP kinases have also been implicated in signal transduction pathways. However, the corresponding role by plant derived GMP kinases remains to be elucidated. Full-length cDNA clones encoding enzymatically active guanylate kinases were isolated from cDNA libraries of lily and tobacco. Lily cDNA is predicted to encode a 392-amino acid protein with a molecular mass of 43.1 kDa and carries amino- and carboxy- terminal extensions of the guanylate kinase (GK)-like domain. But tobacco cDNA is predicted to encode a smaller protein of 297-amino acids with a molecular mass of 32.7 kDa. The amino acid residues known to participate in the catalytic activity of functionally characterized GMP kinases, are also conserved in GK domains of LGK-1 and NGK-1. The GK domains of NGK-1, LGK-1 and previously characterized AGK-1 from Arabidopsis exhibit 74–84% identity, whereas their N- and C-terminal domains are more divergent with amino acid conservation in the order of 48-55%. Phylogenetic analysis on the deduced amino acid sequences reveals that NGK-1 and LGK-1 form one distinct subgroup along with AGK-1 and AGK-2 homologues from Arabidopsis. Isolation of GMP kinases from diverse plant species like lily and tobacco adds a new dimension in understanding their role in cell signaling pathways that are associated with plant growth and development.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007131013675
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  • 2
    Electronic Resource
    Electronic Resource
    Further characterization of the actin-related protein Act3p/Arp4 of S. cerevisiae through mutational analysis (2000)
    Springer
    Molecular biology reports 27 (2000), S. 35-43 
    Details
    ISSN: 1573-4978
    Keywords: ACT3 ; ARP4 ; mutagenesis ; NLS ; nuclear localization ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the course of functional analysis of the yeast ARP4 (ACT3) gene, we further characterized the role of its protein product Arp4p in the cell. We report that although it is localized in the nucleus, Arp4p performs its function independently of binding to DNA directly. The roles of the core actin structure (the ATP-binding pocket) and a putative Nuclear Localization Signal (NLS) of Arp4p were tested by targeted mutagenesis. The results suggest that under normal conditions, the ATPase activity and the NLS are dispensable for the essential function of this protein in the cell. Furthermore, the use of reporter genes confirmed that Arp4p could be involved in some general mechanism of transcriptional regulation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007173530124
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  • 3
    Electronic Resource
    Electronic Resource
    Growth rate of fungi in bathrooms Experimental survey (2000)
    Springer
    Mycoscience 41 (2000), S. 297-301 
    Details
    ISSN: 1618-2545
    Keywords: bathroom ; Cladosporium ; fungal contamination ; Paecilomyces ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Factors promoting fungal contamination of the cement jointing between bathroom tiles were studied in the laboratory. Under continuous wet conditions, the growth of the yeastRhodotolura andCandida on cement was detected from the fourth day of the experiment. Following the rapid growth and decline of the yeast, growth of the moldPaecilomyces was detected on the 12th day. The application of soap or malt extract to the cement promoted the growth ofPaecilomyces. Prolongation of dry conditions delayed the growth of both yeast and mold; under these conditions,Cladosporium, one of the most common molds in household bathrooms, was detected instead ofPaecilomyces. Colonies ofCladosporium were observed along cracks in the cement. On all cement examined, a succession of mycological flora from yeast to mold was found, although fungal genera varied with culture conditions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02463942
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  • 4
    Electronic Resource
    Electronic Resource
    Yeast Mitochondrial Carriers: Bacterial Expression, Biochemical Identification and Metabolic Significance (2000)
    Springer
    Journal of bioenergetics and biomembranes 32 (2000), S. 67-77 
    Details
    ISSN: 1573-6881
    Keywords: Mitochondria ; transport ; overexpression ; dicarboxylate carrier ; ACR1 gene ; succinate-fumarate exchange ; ARG11 gene; ornithine carrier ; arginine biosynthesis ; yeast ; metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract The genome of Saccharomyces cerevisiae encodes 35 members of a family proteins thattransport metabolites and substrates across the inner membranes of mitochondria. They includethree isoforms of the ADP/ATP translocase and the phosphate and citrate carriers. At the startof our work, the functions of the remaining 30 members of the family were unknown. We areattempting to identify these 30 proteins by overexpression of the proteins in specially selectedhost strains of Escherichia coli that allow the carriers to accumulate at high levels in the formof inclusion bodies. The purified proteins are then reconstituted into proteoliposomes wheretheir transport properties are studied. Thus far, we have identified the dicarboxylate,succinate-fumarate and ornithine carriers. Bacterial overexpression and functional identification, togetherwith characterization of yeast knockout strains, has brought insight into the physiologicalsignificance of these transporters. The yeast dicarboxylate carrier sequence has been used toidentify the orthologous protein in Caenorhabditis elegans and, in turn, this latter sequencehas been used to establish the sequence of the human ortholog.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005564429242
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  • 5
    Electronic Resource
    Electronic Resource
    A novel method for immobilization of invertase from the thermophilic fungus, Thermomyces lanuginosus (2000)
    Springer
    World journal of microbiology and biotechnology 16 (2000), S. 151-154 
    Details
    ISSN: 1573-0972
    Keywords: Invertases ; immobilization ; phenyl-Sepharose ; thermophilic fungus ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract An invertase from the thermophilic fungus, Thermomyces lanuginosus was immobilized on phenyl-Sepharose and its properties were studied. Between the soluble and immobilized forms of the invertase, there were not much difference in their optimum pH, K M and V max for sucrose. In contrast, the K M and V max for raffinose changed significantly. The optimum temperature for the immobilized invertase was lower by 10 ∘C. The immobilized invertase showed remarkable stability at 50 ∘C and was less sensitive to inhibition by metal ions. There was no leaching of the enzyme for at least a month when stored in the refrigerator. The method is novel and specific for the thermophilic invertase as a mesophilic invertase (from yeast) did not bind to phenyl-Sepharose.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008901801373
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  • 6
    Electronic Resource
    Electronic Resource
    Evaluation of the maximum specific growth rate of a yeast indicating non-linear growth trends in batch culture (2000)
    Springer
    World journal of microbiology and biotechnology 16 (2000), S. 613-616 
    Details
    ISSN: 1573-0972
    Keywords: Absolute error ; specific growth rate ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The maximum specific growth rate (μmax) of an ethanolic D-xylose-fermenting yeast, Pichia stipitis, showing non-linear growth trends in batch culture, was calculated using the rate equation μ2 = (1/Δt) ln(x 2/x 1). The absolute error Δμ, affecting μ2, was derived using an equation given by Borzani (1994). Based on the assumption of linearity of growth curves between two closest time points, the relation between the two rate formulae, μ1 = (1/x¯)dx t /dt and μ2 = (1/Δt) ln(x 2/x 1) was established. In a particular condition, when μ1 = μ2, an equation has been developed, the roots of which are the specific growth rates at different time points.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008980317225
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  • 7
    Electronic Resource
    Electronic Resource
    Re-examination of some species of the genus Geotrichum Link: Fr (2000)
    Springer
    Antonie van Leeuwenhoek 77 (2000), S. 71-81 
    Details
    ISSN: 1572-9699
    Keywords: DNA heterogeneity ; genome comparison ; Geotrichum ; taxonomy ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nutritional physiology and the growth rate of thirty-four strains representing species of Geotrichum without known teleomorph states were examined. From twenty-seven strains the mol% G+C were calculated from the DNA melting curves. The first derivatives of the melting curves of seven strains, including the type strain of Geotrichum clavatum, demonstrated the presence of two peaks, 12% away from each other; the remaining strains showed only a single broad peak. DNA homology values among strains of the former group were high, indicating their conspecificity. The strains of the latter group could be subdivided into six DNA homology groups, four of which could be identified with recognized species and two may represent novel taxa. A combined key of Geotrichum and its teleomorph states Galactomyces and Dipodascus is presented.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1002030518989
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  • 8
    Electronic Resource
    Electronic Resource
    A novel oxylipin-associated ‘ghosting’ phenomenon in yeast flocculation (2000)
    Springer
    Antonie van Leeuwenhoek 77 (2000), S. 401-406 
    Details
    ISSN: 1572-9699
    Keywords: flocculation ; ghosting ; lectin ; oxylipins ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Research on the distribution of oxylipins (3-hydroxy fatty acids) in flocculant strains of the yeast Saccharomyces cerevisiae led to the uncovering of a novel ‘ghosting’ phenomenon observed during assumed lectin-mediated aggregation. We found that intracellular oxylipin-containing osmiophilic layers migrate through yeast cell walls in a ‘ghostlike’ fashion without visually affecting the cell wall structure or the layers. This migration resulted in the binding of these layers to cell walls of adjacent cells. Consequently, ‘ghosting’ seems a prerequisite for flocculation to occur. However, ‘ghosting’ alone may not be sufficient to ensure flocculation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1002735216303
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  • 9
    Electronic Resource
    Electronic Resource
    Saccharomyces bulderi sp. nov., a yeast that ferments gluconolactone (2000)
    Springer
    Antonie van Leeuwenhoek 77 (2000), S. 223-228 
    Details
    ISSN: 1572-9699
    Keywords: fermentation ; gluconolactone ; raffinose ; Saccharomyces bulderi ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An unknown yeast species was isolated from maize silage and was determined to be novel on the basis of morphological and physiological characteristics, nucleotide sequence of domain D1/D2 of LSU rDNA and from its electrophoretic karyotype. The name for the proposed new species is Saccharomyces bulderi Middelhoven, Kurtzman et Vaughan-Martini (type strain CBS 8638, NRRL Y-27203, DBVPG 7127). S. bulderi is closely related to S. barnettii and S. exiguus from which it can be distinguished by having a double vitamin requirement of biotin and thiamine and by no or slow aerobic growth on raffinose, a sugar that on the contrary is fermented rapidly. Gluconolactone is rapidly fermented with ethanol, glycerol and carbon dioxide being the main products.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1002414301967
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  • 10
    Electronic Resource
    Electronic Resource
    The Ca2+-Transport System of Yeast (Endomyces magnusii) Mitochondria: Independent Pathways for Ca2+ Uptake and Release (2000)
    Springer
    Biochemistry (Moscow) 65 (2000), S. 1352-1356 
    Details
    ISSN: 1608-3040
    Keywords: yeast ; Endomyces magnusii ; mitochondria ; calcium transport ; regulation ; ADP ; NADH ; calcium ions ; Na+-independent calcium release
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Some features of the Ca2+-transport system in mitochondria of the yeast Endomyces magnusii are considered. The Ca2+ uniporter was shown to be specifically activated by low concentrations of physiological modulators such as ADP, NADH, spermine, and Ca2+ itself. The Na+-independent system responsible for Ca2+ release from Ca2+-preloaded yeast mitochondria was characterized. The rate of the Ca2+ release was proportional to the Ca2+ load, insensitive to cyclosporin A and to Na+, inhibited by La3+, TPP+, Pi, and nigericin, while being activated by spermine. We conclude that Ca2+ release from preloaded E. magnusii yeast mitochondria is mediated by a Na+-independent pathway, very similar to that in mitochondria from nonexcitable mammalian tissues. A scheme describing an arrangement of the Ca2+ transport system of yeast mitochondria is proposed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1002840503820
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  • 11
    Electronic Resource
    Electronic Resource
    Interaction of Catalytic Domains in Cytochrome P450scc–Adrenodoxin Reductase–Adrenodoxin Fusion Protein Imported into Yeast Mitochondria (2000)
    Springer
    Biochemistry (Moscow) 65 (2000), S. 1362-1366 
    Details
    ISSN: 1608-3040
    Keywords: cytochrome P450scc ; adrenodoxin ; NADPH:adrenodoxin reductase ; fusion protein ; enzymatic activity ; yeast ; mitochondria ; proteolysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract We have constructed plasmids for yeast expression of the fusion protein pre-cytochrome P450scc–adrenodoxin reductase–adrenodoxin (F2) and a variant of F2 with the yeast CoxIV targeting presequence. Mitochondria isolated from transformed yeast cells contained the F2 fusion protein at about 0.5% of total protein and showed cholesterol hydroxylase activity with 22(R)-hydroxycholesterol. The activity increased 17- or 25-fold when sonicated mitochondria were supplemented with an excess of purified P450scc or a mixture of adrenodoxin (Adx) and adrenodoxin reductase (AdxRed), respectively. These data suggest that, at least in yeast mitochondria, the interactions of the catalytic domains of P450scc, Adx, and AdxRed in the common polypeptide chain are restricted.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1002844604728
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  • 12
    Electronic Resource
    Electronic Resource
    Biochemical characterization of neutral trehalase activity in Saccharomyces boulardii (2000)
    Springer
    World journal of microbiology and biotechnology 16 (2000), S. 691-694 
    Details
    ISSN: 1573-0972
    Keywords: Neutral trehalase ; Saccharomyces boulardii ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Lyophilized cells of the non-pathogenic yeast Saccharomyces boulardii are used in many countries for the treatment of several types of diarrhoea and other gastrointestinal diseases. Although the cells must be viable, their mechanism of action is unknown. The disaccharide trehalose is a protectant against several forms of environmental stress in yeast and is involved in maintaining cell viability. There is no information on the enzymes involved in degradation of trehalose in S. boulardii. The aim of the present study was to characterize trehalase activity in this yeast. Cells of S. boulardii grown in glucose exhibited neutral trehalase activity only in the exponential phase. Acidic trehalase was not detected in glucose medium. Cells grown in trehalose exhibited acid and neutral trehalase activities at all growth stages, particularly in the exponential phase. The optimum pH and temperature values for neutral trehalase activity were determined as 6.5 and 30 °C respectively, the half-life being approximately 3 min at 45 °C. The relative molecular mass of neutral trehalase is 80 kDa and the K m 6.4 mM (±0.6). Neutral trehalase activity at pH 6.5 was weakly inhibited by 5 mM EDTA and strongly inhibited by ATP, as well as the divalent ions Cu++, Fe++ and Zn++. Enzyme activity was stimulated by Mg++ and Ca++ only in the absence of cAMP. The presence of cAMP with no ion additions increased activity by 40%.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008966501012
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  • 13
    Electronic Resource
    Electronic Resource
    Iron uptake by the yeast Pichia guilliermondii. Flavinogenesis and reductive iron assimilation are co-regulated processes (1999)
    Springer
    BioMetals 12 (1999), S. 295-300 
    Details
    ISSN: 1572-8773
    Keywords: iron ; yeast ; flavinogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Pichia guilliermondii cells overproduce riboflavin (vitamin B2) in responce to iron deprivation. The increase in ferrireductase activity in iron-starved P. guilliermondii cells correlated with the increase in flavin excretion. As in Saccharomyces cerevisiae, a typical b-type cytochrome spectrum was associated with the plasma membrane fraction of P. guillermondii and the cell ferrireductase activity was strongly inhibited by diphenylene-iodonium, an inhibitor of flavoproteins, in both yeasts. Mutants of P. guilliermondii with increased ferrireductase activity were selected for further investigation of the relationship between iron reduction/uptake and flavin production. The obtained mutation has been called hit (high iron transport). A hit mutant with a single recessive mutation showed the following phenotype: high ferrireductase activity, increased rate of iron uptake and elevated flavinogenic activity. Cu(II) (50 μm) strongly inhibited the growth of the hit mutant compared to the wild-type. The mutant cells grown in copper-supplemented medium (5–25 μm) showed an increase of the ferrireductase activity (up to 2–3 fold). The copper content of the mutant cells grown under these conditions was also higher (1.5–2 fold) than that of the wild-type. The role of the HIT gene of P. guillermondii in the regulation of iron, copper and flavin metabolisms is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009298530145
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  • 14
    Electronic Resource
    Electronic Resource
    Structure and functional analyses of the 26S proteasome subunits from plants – Plant 26S proteasome (1999)
    Springer
    Molecular biology reports 26 (1999), S. 137-146 
    Details
    ISSN: 1573-4978
    Keywords: Arabidopsis ; Physcomitrella patens ; proteasome ; proteolysis ; ubiquitin ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract As initial steps to define how the 26S proteasome degrades ubiquitinated proteins in plants, we have characterized many of the subunits that comprise the proteolytic complex from Arabidopsis thaliana. A set of 23 Arabidopsis genes encoding the full complement of core particle (CP) subunits and a collection encoding 12 out of 18 known eukaryotic regulatory particle (RP) subunits, including six AAA-ATPase subunits, were identified. Several of these 26S proteasome genes could complement yeast strains missing the corresponding orthologs. Using this ability of plant subunits to functionally replace yeast counterparts, a parallel structure/function analysis was performed with the RP subunit RPN10/MCB1, a putative receptor for ubiquitin conjugates. RPN10 is not essential for yeast viability but is required for amino acid analog tolerance and degradation of proteins via the ubiquitin-fusion degradation pathway, a subpathway within the ubiquitin system. Surprisingly, we found that the C-terminal motif required for conjugate recognition by RPN10 is not essential for in vivo functions. Instead, a domain near the N-terminus is required. We have begun to exploit the moss Physcomitrella patens as a model to characterize the plant 26S proteasome using reverse genetics. By homologous recombination, we have successfully disrupted the RPN10 gene. Unlike yeast rpn10Δ strains which grow normally, Physcomitrella rpn10Δ strains are developmentally arrested, being unable to initiate gametophorogenesis. Further analysis of these mutants revealed that RPN10 is likely required for a developmental program triggered by plant hormones.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006926322501
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  • 15
    Electronic Resource
    Electronic Resource
    Assembly of the Yeast Vacuolar Proton-Translocating ATPase (1999)
    Springer
    Journal of bioenergetics and biomembranes 31 (1999), S. 39-47 
    Details
    ISSN: 1573-6881
    Keywords: yeast ; V-ATPase ; assembly ; vacuoles ; multisubunit ; endoplasmic reticulum ; membrane yeast ; V-ATPase ; assembly ; vacuoles ; multisubunit ; endoplasmic reticulum ; membrane proteins ; proton-translocating
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract The yeast vacuolar proton-translocating ATPase (V-ATPase) is the bestcharacterized member of the V-ATPase family. Biochemical and genetic screensled to the identification of a large number of genes in yeast, designatedVMA, encoding proteins required to assemble a functional V-ATPase. Atotal of thirteen genes encode subunits of the final enzyme complex. Inaddition to subunit-encoding genes, we have identified three genes that codefor proteins that are not part of the final V-ATPase complex yet required forits assembly. We refer to these nonsubunit Vma proteins as assembly factors,since their function is dedicated to assembling the V-ATPase. The assemblyfactors, Vma12p, Vma21p, and Vma22p are localized to the endoplasmicreticulum (ER) and aid the assembly of newly synthesized V-ATPase subunitsthat are translocated into the ER membrane. At least two of these proteins,Vma12p and Vma22p, function together in an assembly complex and interactdirectly with nascent V-ATPase subunits.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005455411918
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  • 16
    Electronic Resource
    Electronic Resource
    Subunit f of the Yeast Mitochondrial ATP Synthase: Topological and Functional Studies (1999)
    Springer
    Journal of bioenergetics and biomembranes 31 (1999), S. 85-94 
    Details
    ISSN: 1573-6881
    Keywords: yeast ; mitochondria ; ATP synthase ; ATP17 gene ; subunit f ; orientation ; cross-linking
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Modified versions of subunit f were produced by mutagenesis of theATP17 gene of Saccharomyces cerevisiae. A version of subunit f devoid of thelast 28 amino acid residues including the unique transmembranous domaincomplemented the oxidative phosphorylation of the null mutant. However, atwo-fold decrease in the specific ATP synthase activity was measured andattributed to a decrease in the stability of the mutant ATP synthase complexas shown by the low oligomycin-sensitive ATPase activity at alkaline pH. Themodification or not by non-permeant maleimide reagents of cysteine residuesintroduced at the N and C termini of subunit f indicated aNin-Cout orientation. From the C terminus of subunit fit was possible to cross-link subunit 4 (also called subunit b), which isanother component of the F0 sector and which also displays a shorthydrophilic segment exposed to the intermembrane space.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005407525915
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  • 17
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    The Role of the Amino-Terminal β-Barrel Domain of the α and β Subunits in the Yeast F1-ATPase (1999)
    Springer
    Journal of bioenergetics and biomembranes 31 (1999), S. 95-104 
    Details
    ISSN: 1573-6881
    Keywords: F1-ATPase ; β-barrel domain ; mitochondria ; assembly ; yeast ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract The crystal structure of mitochondrial F1-ATPase indicatesthat the α and β subunits fold into a structure defined by threedomains: the top β-barrel domain, the middle nucleotide-binding domain,and the C-terminal α-helix bundle domain (Abraham et al.1994); Bianchet et al., 1998). The β-barrel domains of theα and β subunits form a crown structure at the top ofF1, which was suggested to stabilize it (Abraham et al.1994). In this study. the role of the β-barrel domain in the α andβ subunits of the yeast Saccharomyces cerevisiae F1,with regard to its folding and assembly, was investigated. The β-barreldomains of yeast F1 α and β subunits were expressedindividually and together in Escherichia coli. When expressedseperately, the β-barrel domain of the β subunit formed a largeaggregate structure, while the domain of the α subunit waspredominately a monomer or dimer. However, coexpression of the β-barreldomain of α subunit domain. Furthermore, the two domains copurified incomplexes with the major portion of the complex found in a small molecularweight form. These results indicate that the β-barrel domain of theα and β subunits interact specifically with each other and thatthese interactions prevent the aggregation of the β-barrel domain of theβ subunit. These results mimic in vivo results and suggest thatthe interactions of the β-barrel domains may be critical during thefolding and assembly of F1.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005443609985
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  • 18
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    Activation and Deactivation of F0F1-ATPase in Yeast Mitochrondia (1999)
    Springer
    Journal of bioenergetics and biomembranes 31 (1999), S. 105-117 
    Details
    ISSN: 1573-6881
    Keywords: mitochondria ; F0F1 ATPase ; ATP synthase ; ATP hydrolysis ; IF1 ; yeast ; regulation ; inactivation ; proton gradient ; detergent
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract The regulation of membrane-bound proton F0F1ATPase by the protonmotive force and nucleotides was studied in yeastmitochondria. Activation occurred in whole mitochondria and the ATPaseactivity was measured just after disrupting the membranes with Triton X-100.Deactivation occurred either in whole mitochondria uncoupled with FCCP, or indisrupted membranes. No effect of Triton X-100 on the ATPase was observed,except a slow reactivation observed only in the absence of MgADP. BothAMPPNP and ATP increased the ATPase deactivation rate, thus indicating thatoccupancy of nucleotidic sites by ATP is more decisive than catalyticturnover for this process. ADP was found to stimulate the energy-dependentATPase activation. ATPase deactivated at the same rate in uncoupled anddisrupted mitochondria. This suggests that deactivation is not controlled byrebinding of some soluble factor, like IF1, but rather by the conversion ofthe F1.IF1 complex into an inactive form.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005495626823
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  • 19
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    Isolation and characterization of Metolachlor-resistant mutants of Saccharomyces cerevisiae (1999)
    Springer
    World journal of microbiology and biotechnology 15 (1999), S. 679-681 
    Details
    ISSN: 1573-0972
    Keywords: Chloroacetamides ; gene mapping ; herbicide resistance ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The herbicide Metolachlor (α-chloroacetamide group) inhibits the growth of Saccharomyces cerevisiae on complete, minimal, and non-fermentative media. Spontaneous and induced resistant mutants showed monogenic segregation patterns. Among the resistant clones, 70% were recessives, 16.4% were partially dominants and 13.4% were dominants. The spontaneous partially dominant mutation Mtc1 was mapped on linkage group XV at 33.3 cM from ade2 and 31.7 cM from his3, in a region that is characterized by the presence of several resistant genes. The recessive mutation mtc2 was located on chromosome IV. Although all the mutants had the ability to grow in the presence of the herbicide, they remained affected in their respiration efficiency, indicating two different mechanism of action of Metholachor on yeast cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008943914292
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  • 20
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    Detection of yeast by PCR in sucrose solutions using a sample preparation method based on filtration (1999)
    Springer
    World journal of microbiology and biotechnology 15 (1999), S. 345-348 
    Details
    ISSN: 1573-0972
    Keywords: Filtration ; naturally contaminated samples ; PCR ; sample preparation ; sucrose solutions ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract An assay based on PCR was developed for the detection of yeast in naturally contaminated industrial sugar solutions. A number of characterized as well as non-characterized yeast strains, isolated from samples from a sugar refinery, were detected with the PCR assay. Specificity tests showed that the presence of neither bacteria nor mould resulted in false positive results. A concentration step, based on filtration, was employed prior to the PCR detection in order to reach a detection level of 0.1 c.f.u./ml of naturally contaminated sugar solution. The detection method based on PCR was found to be more rapid (〈5 h) and easier to perform than the slower and more labour-intensive, traditional culturing techniques.
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    Interaction of aluminum ion with ATP. Mechanism of the aluminum inhibition of glycerol kinase and its reversal by spermine (1998)
    Springer
    BioMetals 11 (1998), S. 63-67 
    Details
    ISSN: 1572-8773
    Keywords: aluminum ion ; glycerol kinase ; NMR ; spermine ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Aluminum ion inhibited yeast glycerol kinase competitively with respect to the substrate MgATP. The K value of the enzyme for aluminum ion was about 3 μM. Spermine at physiological concentrations prevented glycerol kinase from the inhibition by aluminum ion. Nuclear magnetic resonance spectroscopy showed the specific elimination by spermine of aluminum from the metal-ATP complex, but no dissociation of MgATP complex by spermine. Inhibition by aluminum ion of glycerol kinase as well as hexokinase can reduce the utilization of energy fuel in yeast. Change in polyamine concentration may control energy production in vivo, and is responsible for the development of age-related aluminum toxicity. © Rapid Science 1998.
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    Yeasts in the gastrointestinal tract of preweaned calves and possible involvement of Candida glabrata in neonatal calf diarrhea (1998)
    Springer
    Mycopathologia 141 (1998), S. 7-14 
    Details
    ISSN: 1573-0832
    Keywords: Calves ; Candida glabrata ; diarrhea ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract To examine the possibility of a mycotic involvement in neonatal calf diarrhea (NCD) the presence of fungi was assessed in (a) the intestinal contents of dead calves and fecal samples submitted for routine laboratory examination, (b) fecal specimens, sampled once in winter and once in summer, of calves raised on 2 farms with different management systems, and (c) mucosal scrapings of various segments of the digestive tract of a diarrheic calf, massively shedding Candida glabrata. C. glabrata was the most prevalent fungal species isolated from the routine samples. It was the only fungus which was shed by the calves on the 2 farms, for continuous, more or less prolonged periods, but exclusively in the winter months. Diarrhea and C. glabrata shedding seemed to be associated. C. glabrata colonized the abomasum (the functional equivalent of the monogastric stomach) but not the other segments of the digestive tract of the euthanized calf Based on the findings of this study it seems that while some yeast species may be considered as commensals of the digestive tract of calves, and consequently their isolation from intestinal contents or fecal samples has no clinical significance, others, such as C. glabrata may be involved in enteric pathogenic processes. Moreover, characteristics of the culture, previous chemotherapeutic treatments, the animal's age and possibly climatic conditions should be taken into account before deciding on the fungal isolate's clinical relevance. Determination of mycotic involvement in NCD by routine mycological examination of intestinal contents and fecal samples of diarrheic calves may be useful to avoid unnecessary and potentially harmful antibacterial therapy.
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    Regulation of superoxide dismutase synthesis in Candida albicans (1998)
    Springer
    Mycopathologia 141 (1998), S. 59-63 
    Details
    ISSN: 1573-0832
    Keywords: Candida albicans ; superoxide dismutase ; SOD ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The synthesis of superoxide dismutase [SOD: EC 1.15.1.1] in response to various cultural conditions was examined in Candida albicans, an opportunistic yeast which causes candidiasis in immunosuppressed patients. SOD plays an important role in protecting cells from the oxidative damage of superoxide radicals. Maximum SOD activity was found after 72 hrs of yeast growth. The optimum pH and temperature for the SOD activity were 7 and 40 °, respectively. The major SOD activity was found in the cytosol fraction and the level of extracellular SOD was very low. The enzyme was stimulated to varying degrees by cholic acid, procaine and tocopherol. On the basis of inhibitor studies and other enzyme properties, the isolated enzyme from C. albicans is identified as copper and zinc superoxide dismutase.
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    Dipodascus capitatus, Dipodascus spicifer and Geotrichum clavatum: Genomic characterization (1998)
    Springer
    Antonie van Leeuwenhoek 74 (1998), S. 229-235 
    Details
    ISSN: 1572-9699
    Keywords: Dipodascus capitatus ; D.spicifer ; Geotrichum clavatum ; yeast ; taxonomy ; DNA heterogeneity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The G+C contents of 25 strains of Dipodascus capitatus, Dipodascus spicifer and Geotrichum clavatum were found to be heterogeneous on basis of derivative graphs of the melting profiles. Strains showing similar derivative graphs of the melting curve exhibited high levels of DNA homology (80-100%); strains showing dissimilar derivative graphs exhibited low levels of DNA homology (5 to 45%). Being considered separate taxa on basis of these parameters, D. capitatus, D. spicifer and G. clavatum could be identified by a combination of the key characteristics growth on xylose, cellobiose, salicin and arbutin.
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    Quantitative analysis of some mechanisms affecting the yield of oxidative phosphorylation: Dependence upon both fluxes and forces (1998)
    Springer
    Molecular and cellular biochemistry 184 (1998), S. 35-52 
    Details
    ISSN: 1573-4919
    Keywords: oxidative phosphorylation ; leak ; slip ; almitrine mechanistic change in stoichiometry ; fatty acid ; yeast ; rat liver ; mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The purpose of this work was to show how the quantitative definition of the different parameters involved in mitochondrial oxidative phosphorylation makes it possible to characterize the mechanisms by which the yield of ATP synthesis is affected. Three different factors have to be considered: (i) the size of the different forces involved (free energy of redox reactions and ATP synthesis, proton electrochemical difference); (ii) the physical properties of the inner mitochondrial membrane in terms of leaks (H+ and cations); and finally (iii) the properties of the different proton pumps involved in this system (kinetic properties, regulation, modification of intrinsic stoichiometry). The data presented different situations where one or more of these parameters are affected, leading to a different yield of oxidative phosphorylation. (1) By manipulating the actual flux through each of the respiratory chain units at constant protonmotive force in yeast mitochondria, we show that the ATP/O ratio decreases when the flux increases. Moreover, the highest efficiency was obtained when the respiratory rate was low and almost entirely controlled by the electron supply. (2) By using almitrine in different kinds of mitochondria, we show that this drug leads to a decrease in ATP synthesis efficiency by increasing the H+/ATP stoichiometry of ATP synthase (Rigoulet M et al. Biochim Biophys Acta 1018: 91-97, 1990). Since this enzyme is reversible, it was possible to test the effect of this drug on the reverse reaction of the enzyme i.e. extrusion of protons catalyzed by ATP hydrolysis. Hence, we are able to prove that, in this case, the decrease in efficiency of oxidative phosphorylation is due to a change in the mechanistic stoichiometry of this proton pump. To our knowledge, this is the first example of a modification in oxidative phosphorylation yield by a change in mechanistic stoichiometry of one of the proton pumps involved. (3) In a model of polyunsaturated fatty acid deficiency in rat, it was found that non-ohmic proton leak was increased, while ohmic leak was unchanged. Moreover, an increase in redox slipping was also involved, leading to a complex picture. However, the respective role of these two mechanisms may be deduced from their intrinsic properties. For each steady state condition, the quantitative effect of these two mechanisms in the decrease of oxidative phosphorylation efficiency depends on the values of different fluxes or forces involved. (4) Finally the comparison of the thermokinetic data in view of the three dimensional-structure of some pumps (X-ray diffraction) also gives some information concerning the putative mechanism of coupling (i.e. redox loop or proton pump) and their kinetic control versus regulation of mitochondrial oxidative phosphorylation.
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    Phosphatidate Phosphatase—A Key Enzyme in Glycerolipid Biosynthesis. Studies on the Yeast Enzyme (1998)
    Springer
    The protein journal 17 (1998), S. 1-7 
    Details
    ISSN: 1573-4943
    Keywords: Phosphatidate phosphatase ; purification ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Phosphatidate phosphatase is an important enzyme in glycerolipid biosynthesis, but difficult to purify. A purified preparation of N-ethylmaleimide-sensitive phosphatidate phosphatase from the yeast Saccharomyces cerevisiae was obtained by a five-step protocol, using chromatography on DE-53/DEAE FF, Affi-Gel Blue, hydroxyapatite, Mono-Q, and Superdex 200. A protease-deflcient yeast strain gave preparations similar to those of the wild-type strain. In exclusion chromatography, the enzyme activity of all preparations eluted at approximately the same position as albumin. However, the behavior on SDS/PAGE differed considerably among preparations, suggesting a multimeric subunit structure or degradation during purification. A 35-kDa and a 40-kDa protein band which coincided with activity were found in all preparations. Glycerol in the buffers could be excluded without rapid loss of enzyme activity, and Tris could be substituted for ammonium bicarbonate, while at least 0.6% sodium cholate in the buffers was essential.
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    Isolation of an extragenic suppressor of the rna1-1 mutation in Saccharomyces cerevisiae (1998)
    Springer
    Molecular genetics and genomics 259 (1998), S. 404-413 
    Details
    ISSN: 1617-4623
    Keywords: Key wordsrna1-1 ; SRN10 ; RanGAP1 ; Nuclear transport ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The small GTPase Ran is essential for nucleocytoplasmic transport of macromolecules. In the yeast Saccharomyces cerevisiae, Rna1p functions as a Ran-GTPase activating protein (RanGAP1). Strains carrying the rna1-1 mutation exhibit defects in nuclear transport and, as a consequence, accumulate precursor tRNAs. We have isolated two recessive suppressors of the rna1-1 mutation. Further characterization of one of the suppressor mutations, srn10-1, reveals that the mutation (i) can not bypass the need for Rna1p function and (ii) suppresses the accumulation of unspliced pre-tRNA caused by rna1-1. The SRN10 gene is not essential for cell viability and encodes an acidic protein (pI = 5.27) of 24.8 kDa. Srn10p is located in the cytoplasm, as determined by indirect immunofluorescence microscopy. Two-hybrid analysis reveals that there is a physical interaction between Srn10p and Rna1p in vivo. Our results identify a protein that interacts with the yeast RanGAP1.
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    The influence of food quality on the phototactic behaviour of Daphnia magnaStraus (1998)
    Springer
    Hydrobiologia 379 (1998), S. 199-206 
    Details
    ISSN: 1573-5117
    Keywords: zooplankton ; Daphnia ; phototaxis ; food quality ; yeast ; maternal effects ; life history
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We examined the influence of food quality on the phototactic behaviour of Daphnia magna. Cohorts of a positively phototactic D. magna clone were fed nine diets differing in quality. Diets were obtained by substitution of a fraction of unicellular algae ( Scenedesmus acutus) by a biomass-equivalent of fresh yeast ( Saccharomyces cerevisiae). Some of the diets were enriched with an inoculum of ciliates and organic compounds added as a hay infusion filtrate. Animals fed with a diet containing at least 25% algae showed a similar phototactic behaviour as animals fed with a diet that contained 100% algae. Addition of ciliates to the yeast diets resulted in a lower mortality and a higher reproductive rate compared to diets without a ciliate supplement. The presence of ciliates did not influence phototactic behaviour. Experiments testing for a maternal effect showed that the phototactic behaviour of the animals was strongly influenced by diet during the early developmental stages, but was not influenced by the diet of the mother.
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    Analysis of the isopentenyl diphosphate isomerase gene family from Arabidopsis thaliana (1998)
    Springer
    Plant molecular biology 36 (1998), S. 323-328 
    Details
    ISSN: 1573-5028
    Keywords: isoprenoid biosynthetic pathway ; functional complementation ; isopentenyl diphosphate isomerase ; higher plants ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two Arabidopsis thaliana cDNAs (IPP1 and IPP2) encoding isopentenyl diphosphate isomerase (IPP isomerase) were isolated by complementation of an IPP isomerase mutant strain of Saccharomyces cerevisiae. Both cDNAs encode enzymes with an amino terminus that may function as a transit peptide for localization in plastids. At least 31 amino acids from the amino terminus of the IPP1 protein and 56 amino acids from the amino terminus of the IPP2 protein are not essential for enzymatic activity. Genomic DNA blot analysis confirmed that IPP1 and IPP2 are derived from a small gene family in A. thaliana. Based on northern analysis expression of both cDNAs occurs predominantly in roots of mature A. thaliana plants grown to the pre-flowering stage.
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    Binding of Rat Brain Hexokinase to Recombinant Yeast Mitochondria: Effect of Environmental Factors and the Source of Porin (1998)
    Springer
    Journal of bioenergetics and biomembranes 30 (1998), S. 245-255 
    Details
    ISSN: 1573-6881
    Keywords: Mitochondria ; rat brain hexokinase ; porin ; VDAC ; yeast ; cooperative binding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Heterologous binding of rat brain hexokinase to wild type, porinless, and recombinant yeast mitochondria expressing human porin was assessed, partially characterized, and compared to that in the homologous system (rat liver mitochondria). With porin-containing yeast mitochondria it is shown that (i) a significant, saturatable association occurs; (ii) its extent and apparent affinity, correlated with the origin of porin, are enhanced in the presence of dextran; (iii) the binding requires Mg ions and apparently follows a complex cooperative mechanism. This heterologous association does not seem to differ fundamentally from that in the homologous system and represents a good basis for molecular studies in yeast. With porinless yeast mitochondria, binding occurs at much lower affinity, but to many more sites per mitochondrion. The results indicating a major but not exclusive role for porin in the binding are discussed in terms of (i) the mode and mechanism of binding, and (ii) the suitability of the rat hexokinase–yeast mitochondria couple for the study of heterogeneous catalysis in reconstituted cellular model systems.
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    Minireview: Characterization of the Yeast Mitochondria Unselective Channel: A Counterpart to the Mammalian Permeability Transition Pore? (1998)
    Springer
    Journal of bioenergetics and biomembranes 30 (1998), S. 419-429 
    Details
    ISSN: 1573-6881
    Keywords: Permeability transition ; inner mitochondrial membrane ; nucleotides ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Large and unselective permeabilities through the inner membrane of yeast mitochondria have been observed for more than 20 years, but the characterization of these permeabilities, leading to hypothesize the existence of a large-conductance unselective channel in yeast inner mitochondrial membrane, was done only recently by several groups. This channel has been tentatively identified as a yeast counterpart to the mammalian permeability transition pore, the crucial role of which is now well-documented in physiopathological phenomena, such as Ca2+ homeostasis, ischemic damages, or programmed cell death. The aim of this review is to make a point on the known characteristics of this yeast mitochondrial unselective channel (YMUC) and to analyze whether or not it can be considered as a “yeast permeability transition pore.”
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    Attraction of a Flying Nitidulid (Carpophilus humeralis) to Volatiles Produced by Yeasts Grown on Sweet Corn and a Corn-Based Medium (1998)
    Springer
    Journal of chemical ecology 24 (1998), S. 1217-1239 
    Details
    ISSN: 1573-1561
    Keywords: Sap beetle ; attraction ; yeast ; volatile ; headspace ; fermentation ; glucose ; maltose ; sucrose ; maize ; sweet corn
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Attraction to microbial volatiles was examined for the sap beetle, Carpophilus humeralis, which is a pest of maize. Using 54 pure yeast and bacterial cultures, we evaluated differences in volatile emissions among species of microorganisms and whether these differences were associated with insect attraction. On a sterile corn-based medium, both yeasts and bacteria generally multiplied well and produced detectable volatile metabolites. The yeasts ranged from inactive to highly attractive, but no bacterial cultures attracted beetles above control levels. A variety of alcohols, esters, ketones, acids, and phenolic compounds were identified from the headspace above yeast cultures. Growth, volatile production, and, ultimately, attractiveness to beetles depend strongly on the ability of the yeasts to assimilate and/or ferment the carbohydrates present. Abundant volatile production on sweet corn was observed only with yeasts that are able to ferment sucrose and/or maltrose. Saccharomyces cerevisiae (ferments glucose, sucrose, and maltose) and Candida shehatae (ferments glucose and maltose) produced considerably more attractive volatiles than Candida guilliermondii, which only ferments glucose. Yeast volatiles important for beetle attraction included typical fermentation-associated substances (ethanol, acetaldehyde, 2-methyl-1-propanol, 1-propanol, ethyl acetate, 3-methyl-1-butanol and 2-methyl-1-butanol), and also 3-hydroxy-2-butanone, whose presence was not correlated with the occurrence of fermentation. Using aqueous mixtures of synthetic components that produced headspace compositions simulating those of attractive yeasts, it was shown that the typical fermentation volatiles are attractive but not essential for attractiveness. 3-Hydroxyl-2-butanone is sufficient but not necessary, although its attractiveness is enhanced by the presence of fermentation volatiles such as ethanol and 2-methyl-1-proponol. In nature, the beetles could take advantage of a variety of different microbial metabolic processes to locate hosts. The laboratory bioaasays in this study involved flight and therefore were particularly relevant to host-finding behavior in the field.
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    Physiological characterisation of a pyruvate-carboxylase-negative Saccharomyces cerevisiae mutant in batch and chemostat cultures (1998)
    Springer
    Antonie van Leeuwenhoek 74 (1998), S. 253-263 
    Details
    ISSN: 1572-9699
    Keywords: Saccharomyces cerevisiae ; pyruvate carboxylase ; anaplerotic reactions ; sugar metabolism ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A prototrophic pyruvate-carboxylase-negative (Pyc-) mutant was constructed by deleting the PYC1 and PYC2 genes in a CEN.PK strain of Saccharomyces cerevisiae. Its maximum specific growth rate on ethanol was identical to that of the isogenic wild type but it was unable to grow in batch cultures in glucose-ammonia media. Consistent with earlier reports, growth on glucose could be restored by supplying aspartate as a sole nitrogen source. Ethanol could not replace aspartate as a source of oxaloacetate in batch cultures. To investigate whether alleviation of glucose repression allowed expression of alternative pathways for oxaloacetate synthesis, the Pyc- strain and an isogenic wild-type strain were grown in aerobic carbon-limited chemostat cultures at a dilution rate of 0.10 h-1 on mixtures of glucose and ethanol. In such mixed-substrate chemostat cultures of the Pyc- strain, steady-state growth could only be obtained when ethanol contributed 30% or more of the substrate carbon in the feed. Attempts to further decrease the ethanol content of the feed invariably resulted in washout. In Pyc- as well as in wild-type cultures, levels of isocitrate lyase, malate synthase and phospho-enol-pyruvate carboxykinase in cell extracts decreased with a decreasing ethanol content in the feed. Nevertheless, at the lowest ethanol fraction that supported growth of the Pyc- mutant, activities of the glyoxylate cycle enzymes in cell extracts were still sufficient to meet the requirement for C4-compounds in biomass synthesis. This suggests that factors other than glucose repression of alternative routes for oxaloacetate synthesis prevent growth of Pyc-mutants on glucose.
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    Validation of the basidiomycetous yeast, Sporidiobolus microsporus sp. nov., based on phenotypic and molecular analyses (1998)
    Springer
    Antonie van Leeuwenhoek 74 (1998), S. 265-270 
    Details
    ISSN: 1572-9699
    Keywords: basidiomycete ; nucleotide sequence analysis ; Sporidiobolus microsporus ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The validation of Sporidiobolus microsporus Higham, nom. nud. is based on phenotypic characterization and molecular sequence analysis of a partial region of the large sub-unit ribosomal DNA. The species is compared, based on phenotypic and molecular characteristics, with other species of Sporidiobolus and the closely related Rhodosporidium fluviale.
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    Microbiological treatment of industrial wastes containing toxic chromium involving successive use of bacteria, yeast and algae (1998)
    Springer
    World journal of microbiology and biotechnology 14 (1998), S. 583-585 
    Details
    ISSN: 1573-0972
    Keywords: Algae ; bacteria ; chromium resistance ; reduction of chromium ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Bacteria, yeasts, protozoa and algae, observed in industrial effluents from tanneries, were checked for their possible use in the detoxification of polluted water. Two bacterial strains were found to be highly resistant to CrVI. Several bacteria, yeasts and algae were observed to be capable of reducing CrVI to CrIII.
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    Review: Ethanol production at elevated temperatures and alcohol concentrations: Part I – Yeasts in general (1998)
    Springer
    World journal of microbiology and biotechnology 14 (1998), S. 809-821 
    Details
    ISSN: 1573-0972
    Keywords: Alcohol ; ethanol ; thermophilic ; thermotolerant ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract There are a number of process advantages which could be exploited through the use of thermophilic microorganisms for ethanol production. Energy savings through reduced cooling costs, higher saccharification and fermentation rates, continuous ethanol removal and reduced contamination have stimulated a search for routes to thermophilic or thermotolerant yeasts. These routes have included screening existing culture collections, temperature adaptation, mutagenesis and molecular techniques and finally isolating new strains. Varying success has been achieved, however, the most thermotolerant yeasts have come from fresh isolations from environments which experience high temperatures. Thermotolerant yeasts have been investigated for the following potential applications: simultaneous saccharification and fermentation of cellulose, where the high fermentation temperature allows more rapid and efficient enzymatic cellulose hydrolysis; whey fermentation, where high salt and low fermentable substrate concentrations make conditions difficult; and fermentation of D-xylose and cellobiose, which is essential for efficient conversion of woody biomass to ethanol. Ethanol and temperature tolerance are important characteristics for commercial yeast strains. Both characteristics are interactive and generally decrease with increasing temperature and ethanol concentration. Considerable research has been directed towards investigation of fatty acid composition changes in response to these stresses and the role of heat shock proteins in tolerance mechanisms. If thermotolerant yeasts are to be used in commercial processes, bioreactor configuration will play an important part in the design of production processes. Batch and fed-batch systems have been shown to be useful in some circumstances as have continuous flow systems, however, some of the newly isolated thermotolerant yeasts such as Kluyveromyces marxianus do not show the high growth rate under anaerobic conditions that is characteristic of Saccharomyces cerevisiae. Various immobilization techniques appear to offer a means of presenting and maintaining high biomass in anaerobic continuous flow reactors.
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    Study on effect of diluent osmotic pressure on yeast cell strength and cell size using a novel micromanipulation technique (1998)
    Springer
    World journal of microbiology and biotechnology 14 (1998), S. 719-725 
    Details
    ISSN: 1573-0972
    Keywords: Coulter counter ; mechanical properties ; micromanipulation ; osmotic pressure ; Saccharomyces cerevisiae ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A new micromanipulation technique which has previously been used to measure the mechanical properties of single animal cells has now been applied to yeast cells. In this study this technique was used to measure yeast cell strength and cell size across a 2l batch fermentation. Alternatively the cell size could also be determined using a Coulter counter while cell measurement was diluted with a conducting fluid (Isoton II). For the cell strength, it was found that the osmotic pressure of diluents did affect cell strength. However, it was also found that there was no significant effect of osmotic pressure of diluents on cell size whether a Coulter counter or micromanipulation was used for measurement. Micromanipulation has been shown to be a powerful technique for measuring the mechanical properties of yeast cells and it will be very useful for studying their behaviour in cell disruption equipment, e.g. high-pressure homogenizers.
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    Inulase-secreting strain of Saccharomyces cerevisiae produces fructose (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 60 (1998), S. 492-497 
    Details
    ISSN: 0006-3592
    Keywords: yeast ; inulin ; inulase ; fructose ; secretion ; hexokinases ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The gene encoding inulase of the yeast Kluyveromyces marxianus (INU1Km) was cloned and expressed in the inulin-negative yeast Saccharomyces cerevisiae. Cells of S. cerevisiae transformed with the INU1Km gene have acquired extracellular inulase activity and were able to grow in the medium with inulin as a sole carbon source. The S. cerevisiae strain was constructed that is capable of heterologous expression of secreted K. marxianus inulase and is defective in fructose uptake due to null-mutations of the hexokinase structural genes HXK1 and HXK2. When grown in inulin-containing media, this strain is capable of accumulating at least 10% glucose-free fructose in the culture liquid. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 60: 492-497, 1998.
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    On-line estimation of biomass through pH control analysis in aerobic yeast fermentation systems (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 58 (1998), S. 445-450 
    Details
    ISSN: 0006-3592
    Keywords: on-line control ; pH control ; growth monitoring ; proton titration ; yeast ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The amount of acid or base consumed in yeast cultures has been recently assigned to the pathway of nitrogen assimilation under respiratory conditions with no contribution by carbon metabolism (Castrillo et al., 1995). In this investigation, experiments under respirofermentative conditions have shown that production or consumption of ethanol does not contribute significantly to the specific rate of proton production (qH+), thus extending the previously obtained relationships for all aerobic conditions in which other major acid/base contributions are not involved. Tests in batch and chemostat culture confirm the validity of qH+ as a formal control parameter in aerobic fermentations. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:445-450, 1998.
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    Initiation of DNA replication in eukaryotic chromosomes This article is a U.S. Government work and, as such, is in the public domain in the United States of America. (1998)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 72 (1998), S. 8-17 
    Details
    ISSN: 0730-2312
    Keywords: eukaryote ; DNA replication ; replication origin ; pre-replication complex ; initiation proteins ; origin recognition complex ; DNA unwinding ; nuclear structure ; chromatin structure ; DNA methylation ; animal development ; metazoa ; mammal ; frog ; fly ; yeast ; Xenopus ; Drosophila ; Sciara ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Our understanding of the process by which eukaryotes regulate initiation of DNA replication has made remarkable advances in the past few years, thanks in large part to the explosion of genetic and biochemical information on the budding yeast, Saccharomyces cerevisiae. At least three major concepts have emerged: 1) The sequence of molecular events that determines when replication begins and how frequently each replication site is used are conserved among most, if not all, eukaryotes; 2) specific replication origins are used in most, if not all, eukaryotes that consist of a flexible modular anatomy; and 3) epigenetic factors such as chromatin structure and nuclear organization determine which of many potential replication origins are used at different stages in animal development. Thus, the current state of our knowledge suggests a simple unifying concept - all eukaryotes utilize the same basic proteins and DNA sequences to initiate replication, but the metazoa can change both the number and locations of replication origins in response to the demands of animal development. J. Cell. Biochem. Suppls. 30/31:8-17, 1998.
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    Nonflocculent versus total biomass ratio as a criterion for starting the biomass separation process (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 59 (1998), S. 647-650 
    Details
    ISSN: 0006-3592
    Keywords: biomass separation ; flocculation ; biomass measurement ; yeast ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We introduce the ratio of nonflocculent versus total biomass as a criterion for starting cell separation from the medium. This criterion can be applied for the automation of the process regardless of the process dynamics. Its minimum indicates the optimum period of time for the start of the separation process with regard not only to nonflocculent cell concentration, but also medium attributes. In contrast to the concentration of nonflocculent cells, which has two minima, first at the beginning of the process and another broader one in the period during which maximum flocculation is present, the ratio has a single minimum and can therefore be implemented as a criterion for cell separation. To calculate the ratio value, in addition to an on-line method for nonflocculent biomass measurement described elsewhere, an on-line method for the total biomass of flocculent yeast is proposed. It is based on the absorbency measurement of the cell biomass, previously deflocculated by EDTA. Therefore, it can be applied in bioprocesses with transparent media and yeast that can be deflocculated by EDTA. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:647-650, 1998.
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    A structured approach for selection among candidate metabolic network models and estimation of unknown stoichiometric coefficients (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 58 (1998), S. 133-138 
    Details
    ISSN: 0006-3592
    Keywords: metabolic modeling ; model selection ; parameter estimation ; identification ; yeast ; stoichiometry ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A metabolic network model is one of the cornerstones of the emerging Metabolic Engineering methodology. In this article, special attention is therefore, given to the phase of model building. A five-stage structured approach to metabolic network modeling is introduced. The basic steps are: (1) to collect a priori knowledge on the reaction network and to build candidate network models, (2) to perform an a priori check of the model, (3) to estimate the unknown parameters in the model, (4) to check the identified model for acceptability from a biological and thermodynamic point of view, and (5) to validate the model with new data. The approach is illustrated with a growth system involving baker's yeast growing on mixtures of substrates. Special attention is given to the central uncertainties in metabolic network modeling, i.e., estimation of energetic parameters in the network and the choice of the source of anabolic reducing equivalents NADPH. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:133-138, 1998.
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    Ultra-wide band electromagnetic radiation does not affect UV-induced recombination and mutagenesis in yeast (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Bioelectromagnetics 19 (1998), S. 128-130 
    Details
    ISSN: 0197-8462
    Keywords: UWB ; recombination ; mutagenesis ; yeast ; ultraviolet light ; Life and Medical Sciences ; Occupational Health and Environmental Toxicology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Physics
    Notes: Cell samples of the yeast Saccharomyces cerevisiae were exposed to 100 J/m2 of 254 nm ultraviolet (UV) radiation followed by a 30 min treatment with ultra-wide band (UWB) electromagnetic pulses. The UWB pulses (101-104 kV/m, 1.0 ns width, 165 ps rise time) were applied at the repetition rates of 0 Hz (sham), 16 Hz, or 600 Hz. The effect of exposures was evaluated from the colony-forming ability of the cells on complete and selective media and the number of aberrant colonies. The experiments established no effect of UWB exposure on the UV-induced reciprocal and non-reciprocal recombination, mutagenesis, or cell survival. Bioelectromagnetics 19: 128-130, 1998. © 1998 Wiley-Liss, Inc.
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    Purification and properties of polyphosphatase from Saccharomyces cerevisiae cytosol (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 383-390 
    Details
    ISSN: 0749-503X
    Keywords: polyphosphatase ; cytosol ; yeast ; purification ; kinetic model ; Mg2+ ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A homogenous polyphosphatase preparation was obtained from Saccharomyces cerevisiae cytosol with a 3·8% yield and 3540-fold purification. The enzyme hydrolysed polyphosphate (polyP) with various chain lengths, including polyP3, and split Pi off the end of the chain. It was inactive with respect to ATP, PPi, and p-nitrophenylphosphate. Its specific activity with polyP15 was 283 U/mg protein. The polyphosphatase was a monomeric protein with a molecular mass of 40 kDa. This enzyme was inactive without divalent cations and with Cu2+ and Ca2+. The ability of other divalent cations to activate the enzyme decreased in the following order: Co2+〉Mn2+〉Mg2+〉Zn2+. A kinetic model of the hydrolysis of polyP3 and action of Mg2+ is proposed. © 1998 John Wiley & Sons, Ltd.
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    Isolation and characterization of Kluyveromyces marxianus mutants deficient in malate transport (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 401-407 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; Kluyveromyces marxianus ; malic acid transport ; mutants ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In malic acid-grown cells of the strains ATCC 10022 and KMS3 of Kluyveromyces marxianus the transport of malic acid occurred by a malate-proton symport, which accepted l-malic, d-malic, succinic and fumaric acids, but not tartaric, malonic or maleic acids. The system was inducible and subjected to glucose repression. Mutants of the strain KMS3, unable to grow in a medium with malic acid, were isolated and checked for their capacity to utilize several carbon sources and to transport dicarboxylic acids by the malate-proton symport. Two distinct clones affected on malate transport were obtained. Both were able to grow on a medium with glycerol or ethanol but not with dl-malic, succinic, oxoglutaric and oxaloacetic acids as the sole carbon and energy sources. However, while one of the mutants (Mal7) displayed activity levels for the enzymes malate dehydrogenase, isocitrate lyase, and phosphoenolpyruvate carboxykinase similar to those of the wild strain, in the other mutant type (Mal6) the activities for the same enzymes were significantly reduced. Plasma membranes from derepressed cells of the wild strain and of the mutants Mal6 and Mal7 were isolated and the protein analysed by SDS-PAGE. The electrophoretic patterns of these preparations differed in a polypeptide with an apparent molecular mass of about 28 kDa, which was absent only in the mutant Mal7. The results indicated that Mal7 can be affected in a gene that encodes a malate carrier in K. marxianus. © 1998 John Wiley & Sons, Ltd.
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    A highly conserved intraspecies homolog of the Saccharomyces cerevisiae elongation factor-3 encoded by the HEF3 gene (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 1105-1113 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; elongation factor-3 ; EF-3 ; homolog ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A paralog (intraspecies homolog) of the Saccharomyces cerevisiae YEF3 gene, encoding elongation factor-3, has been sequenced in the course of the yeast genome project, and identified by database searching; this gene has been designated HEF3. Bioinformatic and Northern blot analysis indicate that the HEF3 gene is not expressed during vegetative growth. Deletion of the HEF3 gene reveals no growth defects, nor any defects in mating or sporulation. A high copy 2μ clone of HEF3 was constructed, and was shown to be unable to complement a null allele of yef3. Finally, an in vitro assay for ribosome-stimulated ATPase activity was performed with isogenic HEF3 and Δhef3 strains; no difference in biochemical activity could be detected in these strains. From these results, we conclude that the HEF3 gene does not encode a functional homolog of YEF3. © 1998 John Wiley & Sons, Ltd.
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    A spheroplast rate assay for determination of cell wall integrity in yeast (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 1159-1166 
    Details
    ISSN: 0749-503X
    Keywords: cell walls ; protease ; β-glucanase ; lysis ; yeast ; antifungal drugs ; glucan ; mannoprotein ; S. cerevisiae ; C. albicans ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The rate of formation of spheroplasts of yeast can be used as an assay to study the structural integrity of cell walls. Lysis can be measured spectrophotometrically in hypotonic solution in the presence of Zymolyase, a mixture of cell wall-digesting enzymes. The optical density of the cell suspension decreases as the cells lyse. We optimized this assay with respect to enzyme concentration, temperature, pH, and growth conditions for several strains of Saccharomyces cerevisiae. The level of variability (standard deviation) was 1-5% between trials where the replications were performed on the same culture using enzyme prepared from the same lot, and 5-15% for different cultures of the same strain. This assay can quantitate differences in cell wall structure (1) between exponentially growing and stationary phase cells, (2) among different S. cerevisiae strains, (3) between S. cerevisiae and Candida albicans, (4) between parental and mutated lines, and (5) between drug- or chemically-treated cells and controls. © 1998 John Wiley & Sons, Ltd.
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    Fluorescent probing of membrane potential in walled cells: diS-C3(3) assay in Saccharomyces cerevisiae (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 1189-1197 
    Details
    ISSN: 0749-503X
    Keywords: carbocyanine fluorescent probes ; membrane potential ; yeast ; cell wall ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Membrane-potential-dependent accumulation of diS-C3(3) in intact yeast cells in suspension is accompanied by a red shift of the maximum of its fluorescence emission spectrum, λmax, caused by a readily reversible probe binding to cell constituents. Membrane depolarization by external KCl (with or without valinomycin) or by ionophores causes a fast and reproducible blue shift. As the potential-reporting parameter, the λmax shift is less affected by probe binding to cuvette walls and possible photobleaching than, for example, fluorescence intensity. The magnitude of the potential-dependent red λmax shift depends on relative cell-to-probe concentration ratio, a maximum shift (572→582 nm) being found in very thick suspensions and in cell lysates. The potential therefore has to be assessed at reasonably low cell (≤5×106 cells/ml) and probe (10-7 M) concentrations at which a clearly defined relationship exists between the λmax shift and the potential-dependent accumulation of the dye in the cells. The redistribution of the probe between the medium and yeast protoplasts takes about 5 min, but in intact cells it takes 10-30 min because the cell wall acts as a barrier, hampering probe penetration into the cells. The barrier properties of the cell wall correlate with its thickness: cells grown in 0·2% glucose (cell wall thickness 0·175±0·015 μm, n=30) are stained much faster and the λmax is more red-shifted than in cells grown in 2% glucose (cell wall thickness 0·260±0·043 μm, n=44). At a suitable cell and probe concentration and under standard conditions, the λmax shift of diS-C3(3) fluorescence provides reliable information on even fast changes in membrane potential in Saccharomyces cerevisiae. © 1998 John Wiley & Sons, Ltd.
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    Isolation and sequence analysis of the orotidine-5'-phosphate decarboxylase gene (URA3) of Candida utilis. Comparison with the OMP decarboxylase gene family (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 1399-1406 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; Candida utilis ; URA3 ; orotidine 5′-monophosphate decarboxylase ; transformation system ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The URA3 gene of Candida utilis encoding orotidine-5′-phosphate decarboxylase enzyme was isolated by complementation in Escherichia coli pyrF mutation. The deduced amino-acid sequence is highly similar to that of the Ura3 proteins from other yeast and fungal species. An extensive analysis of the family of orotidine-5′-phosphate decarboxylase is shown. The URA3 gene of C. utilis was able to complement functionally the ura3 mutation of Saccharomyces cerevisiae. The sequence presented here has been deposited in the EMBL data library under Accession Number Y12660. © 1998 John Wiley & Sons, Ltd.
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    Monovalent cation fluxes and physiological changes of Debaryomyces hansenii grown at high concentrations of KCl and NaCl (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 1355-1371 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; Debaryomyces ; transport ; physiology ; salt tolerance ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Debaryomyces hansenii showed an increased growth in the presence of either 1m KCl or 1m NaCl and a low acidification of the medium, higher for the cells grown in the presence of NaCl. These cells accumulated high concentrations of the cations, and showed a very fast capacity to exchange either Na+ or K+ for the opposite cation. They showed a rapid uptake of 86 Rb+ and 22 Na+ . 86 Rb+ transport was saturable, with Km and Vmax values higher for cells grown in 1m NaCl. 22 Na+ uptake showed a diffusion component, also higher for the cells grown with NaCl. Changes depended on growth conditions, and not on further incubation, which changed the internal ion concentration. K+ stimulated proton pumping produced a rapid extrusion of protons, and also a decrease of the membrane potential. Cells grown in 1m KCl showed a higher fermentation rate, but significantly lower respiratory capacity. ATP levels were higher in cells grown in the presence of NaCl; upon incubation with glucose, those grown in the presence of KCl reached values similar to the ones grown in the presence of NaCl. In both, the addition of KCl produced a transient decrease of the ATP levels. As to ion transport mechanisms, D. hansenii appears to have (a) an ATPase functioning as a proton pump, generating a membrane potential difference which drives K+ through a uniporter; (b) a K+/H+ exchange system; and (c) a rapid cation/cation exchange system. Most interesting is that cells grown in different ionic environments change their studied capacities, which are not dependent on the cation content, but on differences in their genetic expression during growth. © 1998 John Wiley & Sons, Ltd.
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    Fusion of a fission yeast (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 1529-1566 
    Details
    ISSN: 0749-503X
    Keywords: G protein ; mating ; pheromone ; Schizosaccharomyces pombe ; receptor ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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    Designer deletion strains derived from Saccharomyces cerevisiae S288C: A useful set of strains and plasmids for PCR-mediated gene disruption and other applications (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 115-132 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; yeast ; gene disruption ; S288C ; bacteria-yeast shuttle vectors ; auxotrophic markers ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A set of yeast strains based on Saccharomyces cerevisiae S288C in which commonly used selectable marker genes are deleted by design based on the yeast genome sequence has been constructed and analysed. These strains minimize or eliminate the homology to the corresponding marker genes in commonly used vectors without significantly affecting adjacent gene expression. Because the homology between commonly used auxotrophic marker gene segments and genomic sequences has been largely or completely abolished, these strains will also reduce plasmid integration events which can interfere with a wide variety of molecular genetic applications. We also report the construction of new members of the pRS400 series of vectors, containing the kanMX, ADE2 and MET15 genes. © 1998 John Wiley & Sons, Ltd.
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    Regulation of alcoholic fermentation in batch and chemostat cultures of Kluyveromyces lactis CBS 2359 (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 459-469 
    Details
    ISSN: 0749-503X
    Keywords: Crabtree effect ; yeast ; biomass ; Kluyveromyces lactis ; oxygen ; pyruvate decarboxylase ; regulation ; fermentation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Kluyveromyces lactis is an important industrial yeast, as well as a popular laboratory model. There is currently no consensus in the literature on the physiology of this yeast, in particular with respect to aerobic alcoholic fermentation (‘Crabtree effect’). This study deals with regulation of alcoholic fermentation in K. lactis CBS 2359, a proposed reference strain for molecular studies. In aerobic, glucose-limited chemostat cultures (D=0·05-0·40 h-1) growth was entirely respiratory, without significant accumulation of ethanol or other metabolites. Alcoholic fermentation occurred in glucose-grown shake-flask cultures, but was absent during batch cultivation on glucose in fermenters under strictly aerobic conditions. This indicated that ethanol formation in the shake-flask cultures resulted from oxygen limitation. Indeed, when the oxygen feed to steady-state chemostat cultures (D=0·10 h-1) was lowered, a mixed respirofermentative metabolism only occurred at very low dissolved oxygen concentrations (less than 1% of air saturation). The onset of respirofermentative metabolism as a result of oxygen limitation was accompanied by an increase of the levels of pyruvate decarboxylase and alcohol dehydrogenase. When aerobic, glucose-limited chemostat cultures (D=0·10 h-1) were pulsed with excess glucose, ethanol production did not occur during the first 40 min after the pulse. However, a slow aerobic ethanol formation was invariably observed after this period. Since alcoholic fermentation did not occur in aerobic batch cultures this is probably a transient response, caused by an imbalanced adjustment of enzyme levels during the transition from steady-state growth at μ=0·10 h-1 to growth at μmax. It is concluded that in K. lactis, as in other Crabtree-negative yeasts, the primary environmental trigger for occurrence of alcoholic fermentation is oxygen limitation. © 1998 John Wiley & Sons, Ltd.
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    Biochemistry, cell biology and molecular biology of lipids of Saccharomyces cerevisiae (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 1471-1510 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; S. cerevisiae ; lipids ; phospholipids ; sterols ; sphingolipids ; fatty acids ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The yeast Saccharomyces cerevisiae is a powerful experimental system to study biochemical, cell biological and molecular biological aspects of lipid synthesis. Most but not all genes encoding enzymes involved in fatty acid, phospholipid, sterol or sphingolipid biosynthesis of this unicellular eukaryote have been cloned, and many gene products have been functionally characterized. Less information is available about genes and gene products governing the transport of lipids between organelles and within membranes, turnover and degradation of complex lipids, regulation of lipid biosynthesis, and linkage of lipid metabolism to other cellular processes. Here we summarize current knowledge about lipid biosynthetic pathways in S. cerevisiae and describe the characteristic features of the gene products involved. We focus on recent discoveries in these fields and address questions on the regulation of lipid synthesis, subcellular localization of lipid biosynthetic steps, cross-talk between organelles during lipid synthesis and subcellular distribution of lipids. Finally, we discuss distinct functions of certain key lipids and their possible roles in cellular processes. © 1998 John Wiley & Sons, Ltd.
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    Cloning and characterization of the Hansenula polymorpha homologue of the Saccharomyces cerevisiae PMR1 gene (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 1233-1240 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; PMR1 ; Hansenula polymorpha ; Ca2+-ATPase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A gene homologous to Saccharomyces cerevisiae PMR1 has been cloned in the methylotrophic yeast Hansenula polymorpha. The partial DNA fragment of the H. polymorpha homologue was initially obtained by a polymerase chain reaction and used to isolate the entire gene which encodes a protein of 918 amino acids. The putative gene product contains all ten of the conserved regions observed in P-type ATPases. The cloned gene product exhibits 60·3% amino acid identity to the S. cerevisiae PMR1 gene product and complemented the growth defect of a S. cerevisiae pmr1 null mutant in the EGTA-containing medium. The results demonstrate that the H. polymorpha gene encodes the functional homologue of the S. cerevisiae PMR1 gene product, a P-type Ca2+-ATPase. The DNA sequence of the H. polymorpha homologue has been submitted to GenBank with the Accession Number U92083. © 1998 John Wiley & Sons, Ltd.
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    Physical mapping of chromosomes VII and XV of Saccharomyces cerevisiae at 3·5 kb average resolution to allow their complete sequencing (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 601-616 
    Details
    ISSN: 0749-503X
    Keywords: I-Sce I fragmentation ; yeast ; genome ; cosmids ; colinearity ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The high resolution complete physical maps of chromosomes VII and XV were constructed to form the basis for sequencing these chromosomes as part of the European systematic sequencing programme of the yeast genome, using a unique cosmid library from strain FY1679, and an original top-down mapping strategy involving I-Sce I chromosome fragmentation. A total of 138 and 196 cosmid clones were used to construct the maps for VII and XV, respectively, forming two unique contigs that cover the entirety of chromosomes (1091 kb each), except the telomeric repeats. Colinearity of the cosmid inserts with yeast DNA was verified, and the physical maps were eventually compared with the independently generated genetic maps. © 1998 John Wiley & Sons, Ltd.
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    A family of laboratory strains of Saccharomyces cerevisiae carry rearrangements involving chromosomes I and III (1998)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 14 (1998), S. 551-564 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; S. cerevisiae ; genetics ; karyotypes ; chromosomal rearrangements ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In order to study meiotic segregation of chromosome length polymorphism in yeast, we analysed the progeny of a cross involving two laboratory strains FL100trp and YNN295. Analysis of the parental strains led us to detect an important length polymorphism of chromosomes I and III in FL100trp. A reciprocal translocation involving 80 kb of the left arm of chromosome III and 45 kb of the right arm of chromosome I was shown to be the cause for the observed polymorphism in this strain. The characterization of the translocation breakpoints revealed the existence of a transposition hot-spot on chromosome I: the sequence of the translocation joints on chromosomes I and III suggests that the mechanism very likely involved homologous recombination between Ty2 transposable elements on each chromosome. Analysis of FL100, FL200 and FL100trp ura, which are related to FL100trp, shows that this reciprocal translocation is present in some of the strains of the FL series, whereas the parental strain FL100 does not carry the same rearrangement. We evidenced instead the duplication of 80 kb of chromosome III on chromosome I and a deletion of 45 kb of the right arm of chromosome I in this strain, indicating that secondary events might have taken place and that the strain currently named FL100 is not the common ancestor of the FL series. © 1998 John Wiley & Sons, Ltd.
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    Reduction of vanadate to vanadyl by a strain of Saccharomyces cerevisiae (1997)
    Springer
    BioMetals 10 (1997), S. 239-246 
    Details
    ISSN: 1572-8773
    Keywords: NMR and EPR spectroscopy ; speciation ; transformation ; vanadate ; vanadyl ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Three strains of Saccharomyces cerevisiae, SC-1, DBVPG 6173 and DBVPG 6037, were studied for vanadate resistance in complex Sabouraud medium since they did not thrive in different minimal media (yeast nitrogen base with and without amino acids). The strain SC-1 was resistant up to 16 mm of vanadate, whereas the strains DBVPG 6173 and DBVPG 6037 were inhibited by 8 mm and 4 mm vanadate, respectively. The vanadate resistance in strain SC-1 was constitutive and due to the reduction of this oxyanion to vanadyl, which was detected by EPR spectroscopy and visible spectroscopy. The transformation of vanadate to vanadyl took place during the exponential growth phase; 10 mm of vanadate was reduced to vanadyl outside the cells since the oxyanion was not detected in the cell biomass and only a negligible concentration of vanadyl (25 nmoles mg cells dry weight) was found in the biomass. The other two vanadate-sensitive yeast strains only accumulated vanadate and did not reduce the oxyanion to vanadyl.
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    The incorporation of mannoproteins in the cell wall of S. cerevisiae and filamentous Ascomycetes (1997)
    Springer
    Antonie van Leeuwenhoek 72 (1997), S. 229-237 
    Details
    ISSN: 1572-9699
    Keywords: wall ; mannoproteins ; GPI-anchor ; β1 ; 6-glucan ; PLC ; yeast ; fungi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In yeast, glucanase extractable cell wall proteins are anchored to the plasma membrane at an intermediate stage in their biogenesis via a glycosylphosphatidylinositol (GPI) moiety before they become anchored to the wall glucan via a β1,6-glucan linkage. The mechanism of the membrane processing step of cell wall proteins is not known. Here, we report that Ascomycete filamentous fungi involved in food spoilage such as Aspergillus, Paecilomyces and Penicillium, also contain GPI membrane-anchored proteins some of which are processed by an endogenous phospholipase C activity. Furthermore, similar to the situation in yeast, their cell walls contain mannoproteins which are linked to the glucan backbone through a β1,6-glucan linkage. Interestingly, one mould which contains a significant amount of non covalently linked β1,6-glucosylated cell wall proteins, is much more sensitive towards β1,3-glucanases and membrane perturbing peptides than the others.
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    Identification of proteins that interact with a protein of interest: Applications of the yeast two-hybrid system (1997)
    Springer
    Molecular and cellular biochemistry 172 (1997), S. 67-79 
    Details
    ISSN: 1573-4919
    Keywords: protein interaction ; two-hybrid system ; yeast ; screening ; false positive elimination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The yeast two-hybrid system is a molecular genetic test for protein interaction. Here we describe a step by step procedure to screen for proteins that interact with a protein of interest using the two-hybrid system. This process includes, construction and testing of the bait plasmid, screening a plasmid library for interacting fusion proteins, elimination of false positives and deletion analysis of true positives. This procedure is designed to allow investigators to identify proteins and their encoding cDNAs that have a biologically significant interaction with your protein of interest.
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    Bacterial Overexpression of Putative Yeast Mitochondrial Transport Proteins (1997)
    Springer
    Journal of bioenergetics and biomembranes 29 (1997), S. 541-547 
    Details
    ISSN: 1573-6881
    Keywords: Mitochondria ; transporter ; overexpression ; yeast ; membrane proteins ; crystallization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Thirty-two genes have been identified within the genome of the yeast Saccharomyces cerevisiae which putatively encode mitochondrial transport proteins. We have attempted to overexpress a subset of these genes, namely those which encode mitochondrial transporters of unknown function, and have succeeded in overexpressing 19 of these genes. The overexpressed proteins were then isolated and tested for five well-characterized reconstituted transport activities (i.e., the transport of citrate, dicarboxylates, pyruvate, camitine, and aspartate). Utilizing this approach, we have clearly identified the yeast mitochondrial dicarboxylate transport protein, as well as two additional lower-magnitude transport functions (i.e., tricarboxylate and dicarboxylate transport activities). The implications of these results and the considerations relevant to this approach are discussed.
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    Physiological characterization of a microbial sensor containing the yeast Arxula adeninivorans LS3 (1997)
    Springer
    Antonie van Leeuwenhoek 71 (1997), S. 345-351 
    Details
    ISSN: 1572-9699
    Keywords: Arxula adeninivorans ; microbial sensor ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The yeast Arxula adeninivorans LS3 is a suitable organism for use as part of a microbial sensor. In combination with an amperometric oxygen electrode the sensor offered a possibility for the physiological characterization of this yeast. About 300-400 measurements could be carried out with a single Arxula sensor. The microbial sensor was remarkably stable for over 35 days, when kept at 37 °C during the operation time and at room temperature overnight. The physiological characteristics of Arxula adeninivorans LS3 obtained with the sensor technique were identical to the data obtained with the conventional techniques. However, the sensor technique makes it additionally possible to quantify the physiological data. So the substrates ribose, citric acid, glycerol, oil and benzoate produced signals lower than 10% in comparison to the glucose signal. Fructose, xylose, sucrose, maltose, gentianose, glucosamine, glutamic acid, tryptophan, butyric acid, lauryl acid and propionic acid reached 10-70%, galactose, alanine, glycine, lysine and methionine signals were similar to the glucose signal whereas acetic acid, ethyl alcohol, capron acid, capryl acid and caproic acid reached the highest signals up to 434%.
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    Differences between pectic enzymes produced by laboratory and wild-type strains of Saccharomyces cerevisiae (1997)
    Springer
    World journal of microbiology and biotechnology 13 (1997), S. 711-712 
    Details
    ISSN: 1573-0972
    Keywords: Endopolygalacturonase ; pectic enzymes ; Saccharomyces cerevisiae ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The laboratory strain of S. cerevisiae, IM1-8b, showed pectolytic activity in the presence of either glucose, fructose, or sucrose as the carbon source, but not with galactose. The enzyme activity was rapidly lost with shaking. The optimum pH and temperature for activity were 4.5 and 45°C, respectively. The enzyme was an endopolygalacturonase, since it preferentially hydrolysed pectate over pectin and decreased the viscosity of a 5% polygalacturonic solution by about 30% in 30min producing oligogalacturonic acid and digalacturonic acid as end-products.
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    Fouling effects of yeast culture with antifoam agents on microfilters (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 53 (1997), S. 10-16 
    Details
    ISSN: 0006-3592
    Keywords: microfiltration ; fouling ; yeast ; antifoam agents ; depressurization ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The fouling effects of yeast fermentation broths of Candida utilis in the presence of various commercial antifoam agents (PPG2000, B5600, and G832) up to 4.0 mL/L were studied, using Millipore polyvinylidene fluoride 0.22-μm hydrophilic membranes (GVWP), in a stirred-cell system at 50 kPa and 700 rpm. PPG2000, which has a low value of work of adhesion (Wa of 0.81 mN/m), gave a steady flux of broth of 29 L/(h m2) and was found to have no significant fouling effect on the microfiltration of broth. G832, which has a high Wa, (26.0 mN/m) reduced the flux of the broth to 17 L/(h m2); i.e., by 42% when only 1.0 mL/L was used. However, B5600, which has a Wa of 14.3 mN/m, was found to enhance the flux of broth to 54 L/(h m2); i.e., by 86%, due to the preferential adsorption of the B5600 components onto the hydrophobic cell contents released. These results were reinforced by the depressurization experiments performed with both hydrophilic (GVWP) and hydrophobic (GVHP) membranes, using both young and aged broths. B5600 was found to be the optimum antifoam agent in this study in terms of membrane performance and defoaming efficiency. © 1997 John Wiley & Sons, Inc.
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    Effect of acetaldehyde on Saccharomyces cerevisiae and Zymomonas mobilis subjected to environmental shocks (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 53 (1997), S. 71-78 
    Details
    ISSN: 0006-3592
    Keywords: Zymomonas ; yeast ; acetaldehyde ; ethanol ; stress ; inhibition ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The lag phase of Saccharomyces cerevisiae subjected to a step increase in temperature or ethanol concentration was reduced by as much as 60% when acetaldehyde was added to the medium at concentrations less than 0.1 g/L. Maximum specific growth rates were also substantially increased. Even greater proportional reductions in lag time due to acetaldehyde addition were observed for ethanol-shocked cultures of Zymomonas mobilis. Acetaldehyde had no effect on S. cerevisiae cultures started from stationary phase inocula in the absence of environmental shock and its lag-reducing effects were greater in complex medium than in a defined synthetic medium. Acetaldehyde reacted strongly with the ingredients of complex culture media. It is proposed that the effect of added acetaldehyde may be to compensate for the inability of cells to maintain transmembrane acetaldehyde gradients following an environmental shock. © 1997 John Wiley & Sons, Inc.
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    Kinetics and modeling of GM-CSF production by recombinant yeast in a three-phase fluidized bed bioreactor (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 53 (1997), S. 470-477 
    Details
    ISSN: 0006-3592
    Keywords: fluidized bed bioreactor ; recombinant ; yeast ; kinetics ; modeling ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Continuous production of a recombinant murine granulocyte-macrophage colony-stimulating factor (GM-CSF) by Saccharomyces cerevisiae strain XV2181 (a/a, Trp 1) containing plasmid pαADH2 and immobilized on porous glass beads in a fluidized bed bioreactor was studied. Kinetic models for plasmid stability, cell growth, and protein production in the three-phase fluidized bed bioreactor were developed and used to study the effects of solid loading or cell immobilization on plasmid stability and recombinant protein production. With increasing cell immobilization or solid loading in the bioreactor, plasmid stability and protein production improved significantly. The improvements could be attributed to the decreased θ value, which is the plasmid loss probability during cell division and is an indication of segregational instability of the recombinant cell, and the increased α value, which is the ratio of the specific growth rate of a plasmid-carrying cell to that of a plasmid-free cell and is indicative of competitive stability of the recombinant cell culture. θ decreased from 0.552 to 0.042 and α increased from 0.351 to 0.991 when solid loading in the bioreactor was increased from 5% (v/v) to 33%. The model simulation also showed that the specific growth rate of cells in the bioreactor was lower at higher solid loading. This indicated that there was significant mass transfer limitation, particularly for oxygen transfer, when the total cell density in the bioreactor was high at high solid loading. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 470-477, 1997.
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    Construction and performance of plug-in membrane inlet mass spectrometer for fermentor monitoring (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 54 (1997), S. 535-542 
    Details
    ISSN: 0006-3592
    Keywords: fermentor monitoring ; mass spectrometer ; Pichia stipitis ; carbon dioxide ; yeast ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An in situ sterilizable plug-in membrane inlet mass spectrometer for monitoring dissolved gases and volatiles in fermentors was constructed and tested. The design ensured a minimal distance to be traveled by analyte molecules from the bulk of the fermentation broth to the ionization chamber of the mass spectrometer. Apart from the specific cross talk due to overlapping mass peaks from different compounds, we found that carbon dioxide interfered unspecifically with all the mass peaks of other substances, changing them by the same factor. The interference changed slowly with time and could be positive or negative depending on the history of the mass spectrometer. Also, the general sensitivity of the instrument changed slowly with time. These effects can be neglected or corrected for empirically in short-term measurements. When the fermentor was aerated with a three-component gas mixture including carbon dioxide, a rapid change in the partial pressure of carbon dioxide in the gas mixture gave rise to a transient in the signal of a gas whose partial pressure was kept constant. This effect revealed a transient change in the composition of the gas mixture in the bubbles caused by net import or export of carbon dioxide during equilibration with the new gas mixture. An experimental method to determine the effective partial pressures of gases in the bubbles during steady-state transport of carbon dioxide was designed. The plug-in membrane inlet mass spectrometer was tried as a probe for oxygen and ethanol in an oxystatic culture of the yeast Pichia stipitis. We found that it was possible to keep a steady-state concentration of as little as 0.5 μM throughout the lifetime of the culture. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 535-542, 1997.
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    Determination of cells' water membrane permeability: Unexpected high osmotic permeability of Saccharomyces cerevisiae (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 56 (1997), S. 62-70 
    Details
    ISSN: 0006-3592
    Keywords: osmotic shock ; water permeability ; mixing time constant ; mathematical model ; yeast ; leukemia K562 cells ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Water permeability (Lp), calculated from the volume variations of cells subjected to an osmotic shock, is classically used to characterize cell membrane properties. In this work, we have shown the importance of the kind of mixing reactor used to measure the Lp parameter. A mathematical model including the mixing time constant has been proposed allowing an accurate Lp estimation even though the mixing time constant is higher than the cell time constant obtained in response to a perfect shock. The estimated Lp values of human leukemia K562 cells were found to be the same whatever the mixing time constant. The Lp value of Saccharomyces cerevisiae could not be exactly estimated. However, S. cerevisiae has unexpectedly high water permeability, implying that this yeast may contain water channels in the membrane. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 62-70, 1997.
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    Extremely high frequency electromagnetic fields at low power density do not affect the division of exponential phase Saccharomyces cerevisiae cells (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Bioelectromagnetics 18 (1997), S. 142-155 
    Details
    ISSN: 0197-8462
    Keywords: cell cycle ; electromagnetic field ; nonthermal effects ; yeast ; extremely high frequency ; Life and Medical Sciences ; Occupational Health and Environmental Toxicology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Physics
    Notes: Exponentially growing cells of the yeast Saccharomyces cerevisiae were exposed to electromagnetic fields in the frequency range from 41.682 GHz to 41.710 GHz in 2 MHz increments at low power densities (0.5 μW/cm2 and 50 μW/cm2) to observe possible nonthermal effects on the division of this microorganism. The electronic setup was carefully designed and tested to allow precise determination and stability of the electromagnetic field parameters as well as to minimize possible effects of external sources. Two identical test chambers were constructed in one exposure system to perform concurrent control and test experiments at every frequency step under well-controlled exposure conditions. Division of cells was assessed via time-lapse photography. Control experiments showed that the cells were dividing at submaximal rates, ensuring the possibility of observing either an increase or a decrease of the division rate. The data from several independent series of exposure experiments and from control experiments show no consistently significant differences between exposed and unexposed cells. This is in contrast to previous studies claiming nonthermal effects of electromagnetic fields in this frequency range on the division of S. cerevisiae cells. Possible reasons for this difference are discussed. Bioelectromagnetics 18:142-155, 1997. © 1997 Wiley-Liss, Inc.
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    Molecular Characterization of the Glyceraldehyde-3-phosphate Dehydrogenase Gene of Phaffia rhodozyma (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 1231-1242 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; gpd ; codon usage ; carotenoid ; astaxanthin ; splicing ; phylogeny ; evolution ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The glyceraldehyde-3-phosphate dehydrogenase (GPD; EC1.2.1.12)-encoding gene (gpd) was isolated from a genomic library of Phaffia rhodozyma CBS 6938. Unlike some other eukaryotic organisms the gpd gene is represented by a single copy in P. rhodozyma. The complete nucleotide sequence of the coding, as well as the flanking non-coding regions was determined. The nucleotide sequence of gpd predicted six introns and a polypeptide chain of 339 amino acids. The codon usage in the gpd gene of P. rhodozyma was highly biased and was significantly different from the codon usage in other yeasts. Phylogenetic analysis of different yeasts and filamentous asco- and basidiomycetes gpd sequences indicated that the gpd gene of P. rhodozyma forms a cluster with the corresponding genes of filamentous basidiomycetes. © 1997 John Wiley & Sons, Ltd.
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    In silicio identification of glycosyl-phosphatidylinositol-anchored plasma-membrane and cell wall proteins of Saccharomyces cerevisiae (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 1477-1489 
    Details
    ISSN: 0749-503X
    Keywords: GPI-anchor ; GPI-attachment site ; yeast ; Ascomycetes ; fungi ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Use of the Von Heijne algorithm allowed the identification of 686 open reading frames (ORFs) in the genome of Saccharomyces cerevisiae that encode proteins with a potential N-terminal signal sequence for entering the secretory pathway. On further analysis, 51 of these proteins contain a potential glycosyl-phosphatidylinositol (GPI)-attachment signal. Seven additional ORFs were found to belong to this group. Upon examination of the possible GPI-attachment sites, it was found that in yeast the most probable amino acids for GPI-attachment are asparagine and glycine. In yeast, GPI-proteins are found at the cell surface, either attached to the plasma-membrane or as an intrinsic part of the cell wall. It was noted that plasma-membrane GPI-proteins possess a dibasic residue motif just before their predicted GPI-attachment site. Based on this, and on homologies between proteins, families of plasma-membrane and cell wall proteins were assigned, revealing 20 potential plasma-membrane and 38 potential cell wall proteins. For members of three plasma-membrane protein families, a function has been described. On the other hand, most of the cell wall proteins seem to be structural components of the wall, responsive to different growth conditions. The GPI-attachment site of yeast slightly differs from mammalian cells. This might be of use in the development of anti-fungal drugs. © 1997 John Wiley & Sons, Ltd.
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    The Sequence of a 36·7 kb Segment on the Left Arm of Chromosome IV from Saccharomyces cerevisiae Reveals 20 Non-overlapping Open Reading Frames (ORFs) Including SIT4, FAD1, NAM1, RNA11, SIR2, NAT1, PRP9, ACT2 and MPS1 and 11 New ORFs (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 65-71 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; Saccharomyces cerevisiae ; chromosome IV ; genomic sequencing ; SIT4/PPH1 ; FAD1 ; NAM1/MTF2 ; RNA11 ; SIR2/MAR1 ; NAT1/AAA1 ; PRP9 ; ACT2 ; MPS1/RPK1 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A 36 688 bp fragment from the left arm of chromosome IV of Saccharomyces cerevisiae was sequenced. Sequence analysis identified 20 complete non-overlapping open reading frames (ORFs) of at least 100 amino acids. Nine of these correspond to previously identified and sequenced genes: SIT4/PPH1, FAD1, NAM1/MTF2, RNA11, SIR2/MAR1, NAT1/AAA1, PRP9, ACT2 and MPS1/RPK1. Three ORFs show homology to previously sequenced genes. One ORF exhibits a hypothetical yabO/yceC/yfiI family signature and one has the ATP-dependent helicase signature of the DEAD and DEAH box families. Six ORFs show no appreciable homology to any proteins in the database. One of these is identical to yeast expressed sequence tags and therefore corresponds to an expressed gene. In addition, two partial ORFs and 11 ORFs that are totally internal and are not likely to be functional were detected. The sequence has been submitted to the EMBL data library under Accession Number Z71781. © 1997 by John Wiley & Sons, Ltd.
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    Multidrug-Resistant Transport Proteins in Yeast: Complete Inventory and Phylogenetic Characterization of Yeast Open Reading Frames within the Major Facilitator Superfamily (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 43-54 
    Details
    ISSN: 0749-503X
    Keywords: drug resistance ; transport ; yeast ; MFS ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Screening of the complete genome sequence from the yeast Saccharomyces cerevisiae reveals that 28 open reading frames (ORFs) are homologous to each other and to established bacterial members of the drug-resistant subfamily of the major facilitator superfamily. The phylogenesis of these protein sequences shows that they fall into three major clusters. Cluster I contains 12 ORFs, cluster II contains ten ORFs and cluster III contains six ORFs. Hydropathy analyses indicate that in clusters II and III ORFs, 14 transmembrane spans are predicted whereas only 12 transmembrane spans are predicted in cluster I ORFs.Three ORFs that have known functions as multidrug-resistance pumps in other yeast species such as Schizosaccharomyces pombe (CAR1), Candida albicans (BMRP) or C. maltosa (CYHR), also fall into cluster I. Two S. cerevisiae ORFs of known multidrug-resistance function (ATR1, SGE1) fall into cluster II. Cluster III consists exclusively of ORFs of unknown function but binary sequence comparisons show homology to ORFs from cluster II.Analysis of the multiple alignment for these proteins leads to the identification of characteristic signature sequences for each of the three clusters. © 1997 by John Wiley & Sons, Ltd.
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    The Nucleotide Sequence of a 39 kb Segment of Yeast Chromosome IV: 12 New Open Reading Frames, Nine Known Genes and One Gene for Gly-tRNA (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 163-169 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; genome project ; chromosome IV ; GDH ; SHR3 ; UGA4 ; NHP2 ; HEM3 ; MGT1 ; SHM1 ; ASF2 ; Gly-tRNA ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The complete nucleotide sequence of a 39 090 bp segment from the left arm of yeast chromosome IV was determined. Twenty-one open reading frames (ORFs) longer than 100 amino acids and a Gly-tRNA gene were discovered. Nine of the 21 ORFs (D0892, D1022, D1037, D1045, D1057, D1204, D1209, D1214, D1219) correspond to the previously sequenced Saccharomyces cerevisiae genes for the NAD-dependent glutamate dehydrogenase (GDH), the secretory component (SHR3), the GABA transport protein (UGA4), the high mobility group-like protein (NHP2), the hydroxymethylbilane synthase (HEM3), the methylated DNA protein-cysteine S-methyltransferase (MGT1), a putative sugar transport protein, the Shm1 protein (SHM1) and the anti-silencing protein (ASF2). The inferred amino acid sequences of 11 ORFs show significant similarity with known proteins from various organisms, whereas the remaining ORF does not share any similarity with known proteins. The nucleotide sequence has been entered in the EMBL data Library under the Accession Number X99000.©1997 John Wiley & Sons, Ltd.
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    The Sequence of a Nearly Unclonable 22·8 kb Segment on the Left Arm of Chromosome VII from Saccharomyces cerevisiae Reveals ARO2, RPL9A, TIP1, MRF1, Genes and Six New Open Reading Frames (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 177-182 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; genome sequencing ; chromosome VII ; long-range PCR ; clone instability ; ARO2 ; RPL9A ; TIP1 ; MRF1 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of 22 803 bp on the left arm of chromosome VII was determined by polymerase chain reaction-based approaches to compensate for the unstable character of cosmid clones from this region of the chromosome. The coding density of the sequence is particularly high (more than 83%). Twelve open reading frames (ORFs) longer than 300 bp were found, two of which (at the left side) have been described previously (James et al., 1995) after sequencing of an overlapping cosmid. Four other ORFs correspond to published sequences of the known genes ARO2, RPL9A, TIP1 and MRF1. ARO2 codes for chorismate synthetase, RPL9A for protein L9 of the large ribosomal subunit and MRF1 for a mitochondrial translation release factor. The TIP1 product interacts with Sec20p and is thus involved in transport from endoplasmic reticulum to Golgi. Five of the remaining ORFs have not been identified previously, while the sixth (YGL142c) has been partially sequenced as it lies 5′ upstream of MRF1. These six ORFs are relatively large (between 933 and 3657 nucleotides). YGL146c, YGL142c, YGL140c and YGL139w have no significant homology to any protein sequence presently available in the public databases, but show two, nine, nine and eight putative transmembrane spans, respectively. YGL144c has a serine active site signature of lipases. YGL141w has limited homology to several human proteins, one of which mediates complex formation between papillomavirus E6 oncoprotein and tumor suppressor protein p53. The sequence reported in this paper has been deposited in the EMBL DNA data library under Accession Number X99960.©1997 John Wiley & Sons, Ltd.
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    Histone H1 in Saccharomyces cerevisiae (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 151-161 
    Details
    ISSN: 0749-503X
    Keywords: histone H1 ; nuclear localization ; green fluorescence protein ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The existence of histone H1 in the yeast, Saccharomyces cerevisiae, has long been debated. In this report we describe the presence of histone H1 in yeast. YPL127c, a gene encoding a protein with a high degree of similarity to histone H1 from other species was sequenced as part of the contribution of the Montreal Yeast Genome Sequencing Group to chromosome XVI. To reflect this similarity, the gene designation has been changed to HHO1 (Histone H One). The HHO1 gene is highly expressed as poly A+ RNA in yeast. Although deletion of this gene had no detectable effect on cell growth, viability or mating, it significantly altered the expression of β-galactosidase from a CYC1-lacZ reporter. Fluorescence observed in cells expressing a histone H1-GFP protein fusion indicated that histone H1 is localized to the nucleus.©1997 John Wiley & Sons, Ltd.
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    Sequence Analysis of a 37·6 kbp Cosmid Clone from the Right Arm of Saccharomyces cerevisiae Chromosome XII, Carrying YAP3, HOG1, SNR6, tRNA-Arg3 and 23 New Open Reading Frames, Among Which Several Homologies to Proteins Involved in Cell Division Control and to Mammalian Growth Factors and other Animal Proteins are Found (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 241-250 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; genome sequencing ; chromosome XII ; SNR6 ; YAP3 ; HOG1 ; ZFM1 ; FLO1 ; Arg-tRNA ; flocculation ; TPR motif ; crn ; cell cycle control ; transcriptional factor ; pseudogene ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of 37 639 bp of the right arm of chromosome XII has been determined. Twenty-five open reading frames (ORFs) longer than 300 bp were detected, two of which extend into the flanking cosmids. Only two (L2931 and L2961) of the 25 ORFs correspond to previously sequenced genes (HOG1 and YAP3, respectively). Another ORF is distinct from YAP3 but shows pronounced similarity to it. About half of the remaining ORFs show similarity to other genes or display characteristic protein signatures. In particular, ORF L2952 has striking homology with the probable cell cycle control protein crn of Drosophila melanogaster. L2949 has significant similarity to the human ZFM1 (related to a potential suppressor oncogene) and mouse CW17R genes, though it lacks the carboxy-terminal oligoproline and oligoglutamine stretches encoded by these mammalian genes. The small ORF L2922 is similar to part of the much larger yeast flocculation gene FLO1. Other sequences found in the 37 639 bp fragment are one delta and one solo-sigma element, the tRNA-Arg3 gene, the small nuclear RNA gene SNR6 and three ARS consensus sequences. The nucleotide sequence data reported in this paper are available in the EMBL, GenBank and DDBJ Nucleotide Sequence Databases under the Accession Number X89514. ©1997 John Wiley & Sons, Ltd.
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    The DNA Sequence of Cosmid 14-13b from Chromosome XIV of Saccharomyces cerevisiae Reveals an Unusually High Number of Overlapping Open Reading Frames (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 261-266 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; chromosome XIV ; genome sequencing ; OMP1 ; PSU1 ; MLS1 ; RPC19 ; DBP2 ; CYB5 ; ESBP6 ; H8263 ; AF-9 ; ENL ; TFIIF ; TBF1 ; YHR117w ; YKL221w ; YHR115c ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This work is part of the effort for sequencing chromosome XIV of Saccharomyces cerevisiae. Cosmid 14-13b contains a 37·8 kb insert derived from a partial Sau3A digestion of the genome, cloned into the BamHI site of the vector Pou6. The strategy used for sequencing is based on the fragmentation of the whole cosmid by sonication, followed by shotgun sequencing on an Applied Biosystem DNA sequencer. The clones with inserts corresponding to the vector were identified by dot-blot hybridization, without the need of sequencing. The analysis of the DNA sequence reveals 29 open reading frames (ORFs) longer than 300 bases. Nine ORFs are internal to some other ORFs. Similarity searches against DNA and protein data banks show that six ORFs correspond to already known yeast genes (OMP1, PSU1, MLS1, RPC19, DBP2, CYB5) and one ORF matches the sequence of a putative yeast gene (ESBP6). The cosmid sequence has been submitted to the EMBL data bank under Accession Number Z69382.©1997 John Wiley & Sons, Ltd.
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    Sequence Analysis of a Near-subtelomeric 35·4 kb DNA Segment on the Right Arm of Chromosome VII from Saccharomyces cerevisiae Carrying the MAL1 Locus Reveals 15 Complete Open Reading Frames, Including ZUO1, BGL2 and BIO2 Genes and an ABC Transporter Gene (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 251-259 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; genome sequencing ; chromosome VII ; multiple drug resistance ; oligomycin resistance ; maltose fermentation ; maltase ; α-glucosidase ; MAL1 ; ZUO1 ; BGL2 ; BIO2 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of 35 400 bp at approximately 10 kb from the right telomere of chromosome VII was determined. The segment contains the MAL1 locus, one of the five unlinked loci sufficient for maltose utilization. Until now, each of these loci was considered to contain three genes (for regulator, permease and α-glucosidase), but a fourth gene, presumably an extra α-glucosidase gene, was found at MAL1 adjacent to the usual cluster of three genes. The two glucosidase genes are present in opposite orientation, forming an inverted repeat structure. In addition to the four genes at MAL1, there are 11 complete, non-overlapping open reading frames (ORFs) longer than 300 bp in the sequence presented here. A new ABC transporter gene (YGR281w), required for oligomycin resistance was found (YOR1; Katzman et al., 1995), and the previously sequenced BGL2 (YGR282c), ZUO1 (YGR285c) and BIO2 (YGR286c) genes were located. The sequence of BIO2, a biotin synthetase gene, required substantial correction and the size of Bio2p is 375, rather than 356, amino acids. Two ORFs show rather weak similarities to animal genes: YGR278w to an unknown ORF of Caenorhabditis elegans and YGR284c to the murine Surf-4, a member of a cluster of at least four housekeeping genes. The remaining five ORFs do not encode known functions, but three of these show weak to high similarities to other ORFs in the Saccharomyces cerevisiae genome and one (YGR280c) codes for a particularly lysine-rich protein. The nucleotide sequence has been deposited in the EMBL DNA data library under Accession Number X94332. ©1997 John Wiley & Sons, Ltd.
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    Predacious Yeasts (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 225-232 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; predation ; auxotrophy ; sulphate uptake ; haustorium ; necrotrophic mycoparasitism ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Haustorium-mediated predation was observed in seven yeast species. Arthroascus javanensis, Botryoascus synnaedendrus, Guilliermondella selenospora, Saccharomycopsis fibuligera, and three hitherto unknown species penetrate and kill other yeasts. These yeasts share an unusual requirement for organic sulphur. One isolate recovered from Australian Hibiscus was studied in detail and found to attack a broad range of prey species, including ascomycetous and basidiomycetous yeasts as well as moulds. Predation was most effective when growth was on a solid surface and the medium was poor in complex nutrients. Organic sulphur (exemplified by methionine) was identified as a key factor. It serves as a nutritional benefit to the predator and, depending on the concentration, acts as either an inhibitor of predation or possibly a signal for detection of prey. Sampling of a yeast habitat with a medium selective for selenium-resistant yeasts indicated that auxotrophic and predacious yeasts might be more widespread than anticipated.©1997 John Wiley & Sons, Ltd.
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    The Branched-Chain Amino Acid Permease Gene of Saccharomyces cerevisiae, BAP2, Encodes the High-Affinity Leucine Permease (S1) (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 435-439 
    Details
    ISSN: 0749-503X
    Keywords: membrane transport ; yeast ; uptake kinetics ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The amino acid leucine has been shown previously to be transported into a yeast cell by at least three permeases: the general amino acid permease, a high-affinity permease (S1) and a low-affinity permease (S2). We isolated the gene BAP2 as a multicopy suppressor of the YPD- phenotype of aat1leu2 yeast. BAP2 has been identified previously as encoding an amino acid permease which transports branched-chain amino acids. In order to align the genetic and biochemical studies of leucine uptake we completed a detailed kinetic analysis of yeast strains in which the BAP2 gene was disrupted and compared this to the kinetics of uptake of the parental strain. We demonstrate that BAP2 encodes the high-affinity leucine permease previously called S1. © 1997 John Wiley & Sons, Ltd.
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    The Saccharomyces cerevisiae MFS Superfamily SGE1 Gene Confers Resistance to Cationic Dyes (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 891-902 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; Sacchromyces cerevisiae ; PDR ; MFS ; 10-N-nonyl acridine orange ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A gene from Saccharomyces cerevisiae whose overexpression confers resistance to 10-N-nonyl acridine orange (NAO) has been isolated. This cationic dye binds acidic phospholipids and more specifically cardiolipin (Petit, J. M., Maftah, A., Ratinaud, M. H. and Julien, R. Eur. J. Biochem. 209, 267-273, 1992). The isolated gene was found to be identical to SGE1, a partial multicopy suppressor of the gal11 mutation (Amakasu, H., Suzuki, Y., Nishizawa, M. and Fukasawa, T. Genetics 134, 675-683, 1993), that also confers crystal violet resistance to a supersensitive strain (Ehrenhofer-Murray, A. E., Wurgler, F. E. and Sengstag, C. Mol. Gen. Genet. 244, 287-294, 1994). The data presented in this paper show that the SGE1 gene product, a member of the major facilitator superfamily, confers a pleiotropic drug-resistance phenotype when present in high copy number. The results also demonstrate that Sge1p acts as an extrusion permease whose specificity seems restricted to dye molecules possessing a large unsaturated domain that stabilizes a permanent positive charge such as NAO, crystal violet, ethidium bromide or malachite green. © 1997 John Wiley & Sons, Ltd.
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    Tandemly Repeated 147 bp Elements Cause Structural and Functional Variation in Divergent MAL Promoters of Saccharomyces cerevisiae (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 1135-1144 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; MAL6 ; divergent promoter ; repeats ; nucleosomes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have studied four novel MAL promoters isolated from a single strain of bakers' yeast. Within these promoters we have identified up to five tandem 147 bp repeats located between the MAL UAS region and the MALT TATA box. These repeats strongly reduce MALT (maltose permease) gene expression but only weakly reduce MALS (maltase) gene expression. Insertion of the 147 bp elements into the heterologous CYC1 promoter reduced expression when located between the CYC1 UAS and the TATA box, but not when located upstream of the UAS. We propose that these naturally occurring repeats have evolved as a mechanism to lower the level of MALT expression relative to MALS expression, thus avoiding possible toxic effects associated with over-expression from multiple copies of the permease gene. Accession numbers are: WIG1, U86359; WIG3, U86360; WIG4, U86361; WIG5, U86362. © 1997 by John Wiley & Sons, Ltd.
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    Ribosomal Protein L9 is the Product of GRC5, a Homolog of the Putative Tumor Suppressor QM in S. cerevisiae (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 1155-1166 
    Details
    ISSN: 0749-503X
    Keywords: rpL9 ; QM homolog ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Genes encoding members of the highly conserved QM family have been identified in eukaryotic organisms from yeast to man. Results of previous studies have suggested roles for QM in control of cell growth and proliferation, perhaps as a tumor suppressor, and in energy metabolism. We identified recessive lethal alleles of the Saccharomyces cerevisiae QM homolog GRC5 that increased GCN4 expression when present in multiple copies. These alleles encode truncated forms of the yeast QM protein Grc5p. Using a functional epitope-tagged GRC5 allele, we localized Grc5p to a 60S fraction that contained the large ribosomal subunit. Two-dimensional gel analysis of highly purified yeast ribosomes indicated that Grc5p corresponds to 60S ribosomal protein L9. This identification is consistent with the predicted physical characteristics of eukaryotic QM proteins, the highly biased codon usage of GRC5, and the presence of putative Rap1p-binding sites in the 5′ sequences of the yeast GRC5 gene. © 1997 John Wiley & Sons, Ltd.
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    Inducible Membranes in Yeast: Relation to the Unfolded-Protein-Response Pathway (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 1211-1229 
    Details
    ISSN: 0749-503X
    Keywords: endoplasmic reticulum ; IRE1 ; KAR2 ; phospholipids ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Overproduction of an endoplasmic reticulum (ER)-resident membrane protein (cytochrome P450 52A3) and of a secretory protein (invertase) was used to study the regulation of the luminal ER protein Kar2p under conditions that lead to ER proliferation and secretory overload, respectively. In both cases we found (i) a significant increase of Kar2 protein and mRNA levels, (ii) a transcriptional regulation based on the function of the 22 bp unfolded-protein-response element of the KAR2 promoter and (iii) an essential role of the transmembrane kinase Ire1p for upregulation of KAR2 gene expression. These results show that the same mechanism operates when KAR2 induction is triggered by overproduction of cytochrome P450 or invertase and that this mechanism shares the known features of the unfolded-protein-response pathway. Disruption of the IRE1 gene resulted in a marked decrease of the invertase protein levels produced. In contrast, a functional IRE1 gene was not required to reach high-level production of the integral membrane protein cytochrome P450 52A3. Moreover, IRE1 gene disruption did not prevent P450-induced ER proliferation. We suggest that Ire1p-mediated KAR2 induction is, in the case of cytochrome P450 52A3 overproduction, a process which follows on ER proliferation, thereby monitoring the increase of ER size and adjusting the level of Kar2p accordingly. © 1997 John Wiley & Sons, Ltd.
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    Mapping of ure1, ure2 and ure3 Markers in Fission Yeast (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 1195-1197 
    Details
    ISSN: 0749-503X
    Keywords: Schizosaccharomyces pombe ; mapping ; urease ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The following urease genes of the fission yeast Schizosaccharomyces pombe have been mapped by induced haploidization and tetrad analysis - ure1: chromosome arm III-L; ure2 and ure3: chromosome arm I-R. The previously determined tps19-rad1 interval (11-12 cM) has been increased to 18 cM. A convenient medium for rapidly scoring the ure gene markers of fission yeast was developed. © 1997 John Wiley & Sons, Ltd.
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    Actin-, myosin- and ubiquitin-dependent endocytosis (1996)
    Springer
    Cellular and molecular life sciences 52 (1996), S. 1033-1041 
    Details
    ISSN: 1420-9071
    Keywords: Ubiquitin ; yeast ; Saccharomyces cerevisiae ; Dictyostelium discoideum ; cytoskeleton ; mutants ; endocytosis ; actin ; myosin ; calmodulin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Endocytosis is a general term that is used to describe the internalization of external and plasma membrane molecules into the cell interior. In fact, several different mechanisms exist for the internalization step of this process. In this review we emphasize the work on the actin-dependent pathways, in particular in the yeastSaccharomyces cerevisiae, because several components of the molecular machinery are identified. In this yeast, the analysis of endocytosis in various mutants reveals a requirement for actin, calmodulin, a type I myosin, as well as a number of other proteins that affect actin dynamics. Some of these proteins have homology to proteins in animal cells that are believed to be involved in endocytosis. In addition, the demonstration that ubiquitination of some cell surface molecules is required for their efficient internalization is described. We compare the actin, myosin and ubiquitin requirements for endocytosis with recent results found studying these processes usingDictyostelium discoideum and animal cells.
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    Vacuolar H+-ATPase: From mammals to yeast and back (1996)
    Springer
    Cellular and molecular life sciences 52 (1996), S. 1101-1110 
    Details
    ISSN: 1420-9071
    Keywords: Vacuolar H+-ATPase ; biogenesis ; assembly ; proton pump ; bovine ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Vacuolar H+-adenosine triphosphatase (V-ATPase) is composed of distinct catalytic (V1) and membrane (V0) sectors containing several subunits. The biochemistry of the enzyme was mainly studied in organelles from mammalian cells such as chromaffin granules and clathrin-coated vesicles. Subsequently, mammalian cDNAs and yeast genes encoding subunits of V-ATPase were cloned and sequenced. The sequence information revealed the relation between V- and F-ATPases that evolved from a common ancestor. The isolation of yeast genes encoding subunits of V-ATPase opened an avenue for molecular biology studies of the enzyme. Because V-ATPase is present in every known eukaryotic cell and provides energy for vital transport systems, it was anticipated that disruption of genes encoding V-ATPase subunits would be lethal. Fortunately, yeast cells can survive the absence of V-ATPase by ‘drinking’ the acidic medium. So far only yeast cells have been shown to be viable without an active V-ATPase. In contrast to yeast, mammalian cells may have more than one gene encoding each of the subunits of the enzyme. Some of these genes encode tissue- and/or organelle-specific subunits. Expression of these specific cDNAs in yeast cells may reveal their unique functions in mammalian cells. Following the route from mammals to yeast and back may prove useful in the study of many other complicated processes.
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    Role of the conserved histidines in the Zn module of the copper-activated transcription factors in yeast (1996)
    Springer
    JBIC 1 (1996), S. 560-566 
    Details
    ISSN: 1432-1327
    Keywords: Key words Copper ; metalloregulation ; Ace1 ; Amt1 ; transcription factor ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract  The Amt1 and Ace1 transcription factors from Candida glabrata and Saccharomyces cerevisiae are activated by the formation of a tetracopper thiolate cluster resulting in Cu4Zn1Amt1 complexes. An independent Zn module exists within the N-terminal 40 residues of Amt1 and Ace1. There are two conserved histidyl residues within this module. His25 serves as a Zn(II) ligand for the S3N1 site. Thus, Zn(II) ligands are provided by cysteinyl thiolates of Cys11, Cys14, Cys23 and His25. The Zn module is a stably folded domain that binds group IIb metal ions with high affinity. Mutations in Amt1 and Ace1 genes within codons encoding Zn(II) cysteinyl ligands markedly attenuate the ability of Amt1 and Ace1 to mediate Cu-induced expression of metallothionein genes. Thus, the Zn modules in Amt1 and Ace1 are functionally important. A second histidine, His18, is conserved in the Zn module of these metal-activated factors. His18 does not serve a major structural role, but appears important functionally in Ace1 and Amt1.
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    Ethanol yields during the feeding phase in fed-batch fermentation of sugar-cane blackstrap molasses (1996)
    Springer
    World journal of microbiology and biotechnology 12 (1996), S. 415-416 
    Details
    ISSN: 1573-0972
    Keywords: Ethanol ; fed-batch fermentation ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The efficiency of ethanol yield increased from 61% to 88% of the theoretical value as the filling-up time was approached in fed-batch fermentation with Saccharomyces cerevisiae. Temporary accumulation of ethanol within the yeast cells may explain the above variation.
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    Location of the β-galactosidase of the yeast Kluyveromyces marxianus var. marxianus ATCC 10022 (1996)
    Springer
    Antonie van Leeuwenhoek 69 (1996), S. 357-361 
    Details
    ISSN: 1572-9699
    Keywords: yeast ; Kluyveromyces marxianus ; β-galactosidase ; transport ; lactose ; ONPG
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary During the growth of Kluyveromyces marxianus var. marxianus ATCC 10022 on lactose, peaks of glucose, but not β-galactosidase activity, were detected iroculture medium. Harvested and washed whole cells produced glucose and galactose from lactose, or ortho-nitro-phenol from the chromogenic substrate ortho-nitro-phenyl-β-D-galactopyranoside (ONPG), indicating that β-galactosidase is physically associated with cells. ONPG hydrolysis by whole cells presented a monophasic kinetics (Km 36.6 mM) in lactose exponential growth phase cells, but a biphasic kinetics (Km 0.2 and 36.6 mM) in stationary growth phase cells. Permeabilization with digitonin or disruption of cells from both growth phases led to monosite ONPG hydrolysis (Km 2.2 to 2.5 mM), indicating that β=galactosidase is not located in the periplasm. In addition, the energy inhibitors fluoride or arsenate, as well as the uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP) prevented ONPG hydrolysis by whole cells. These findings indicate that energy coupled transmembrane transport is the rate-limiting step for intracellular ONPG cleavage. The taxonomic and physiologic implications of the exclusive intracellular location of β-galactosidase of K. marxianus var. marxianus ATCC 10022 are discussed.
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    Ballistosporous yeasts found on the surface of plant materials collected in New Zealand (1996)
    Springer
    Antonie van Leeuwenhoek 69 (1996), S. 279-291 
    Details
    ISSN: 1572-9699
    Keywords: Taxonomy ; yeast ; basidiomycete ; Bensingtonia ; Bullera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the present study, strains from the surface of plant materials collected in New Zealand that belong to the genera Bensingtonia and Bullera are classified. One strain of Bensingtonia was assigned to Ben. ingoldii, while the remaining strain was assigned to Ben. naganoensis based on DNA-DNA reassociation experiment. Twenty-one of 28 Bullera strains were assigned to B. alba (11 strains), B. crocea (6 strains) and B. variabilis (4 strains). The remaining seven strains could not be assigned to any previously known species and were described as the new species, B. coprosmaensis (1 strain), B. hannae (1 strain), B. huiaensis (1 strain), B. mrakii (3 strains) and B. unica (1 strain).
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    Analysis of coenzyme Q systems, monosaccharide patterns of purified cell walls, and RAPD-PCR patterns in the genus Kluyveromyces (1996)
    Springer
    Antonie van Leeuwenhoek 70 (1996), S. 67-78 
    Details
    ISSN: 1572-9699
    Keywords: yeast ; Kluyveromyces ; systematics ; taxonony ; coenzyme Q ; Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Analysis of the coenzyme Q system and the monosaccharide pattern of purified cell walls were used for species characterization in the genus Kluyveromyces. All the type strains of the genus possess coenzyme Q-6 and the mannose-glucose (‘Saccharomyces type’) cell wall sugar pattern. With the help of Random Amplified Polymorphic DNA-Polymerase Chain Reaction analysis 17 species were separated: K. aestuarii, K. africanus, K. bacillisporus, K. blattae, K. delphensis, K. dobzhanski, K. lactis (anamorph Candida sphaerica), K. lodderae, K. marxianus (syn. K. fragilis, K. bulgaricus, K. cicerisporus anamorphs Candida macedoniensis, C. pseudotropicalis, C. kefyr), K. phaffii, K. piceae, K. polysporus, K. sinensis, K. thermotolerans (syn. K. veronae, anamorph Candida dattila), K. waltii, K. wickerhamii, K. yarrowii (anamorph Candida tannotolerans). A strain of K. drosophilarum showed with the type strain of K. lactis only 63% similarity. The strain originally described as the type strain of K. cellobiovorus nom. nud. was excluded from the genus (Q-9), and found to be conspecific with the type strain of Candida intermedia.
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    Mammalian CAP interacts with CAP, CAP2, and actin (1996)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 61 (1996), S. 459-466 
    Details
    ISSN: 0730-2312
    Keywords: adenylyl cyclase ; BAT3 ; cytoskeleton ; RAS ; signaling ; yeast ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We previously identified human CAP, a homolog of the yeast adenylyl cyclase - associated protein. Previous studies suggest that the N-terminal and C-terminal domains of CAP have distinct functions. We have explored the interactions of human CAP with various proteins. First, by performing yeast two-hybrid screens, we have identified peptides from several proteins that interact with the C-terminal and/or the N-terminal domains of human CAP. These peptides include regions derived from CAP and BAT3, a protein with unknown function. We have further shown that MBP fusions with these peptides can associate in vitro with the N-terminal or C-terminal domains of CAP fused to GST. Our observations indicate that CAP contains regions in both the N-terminal and C-terminal domains that are capable of interacting with each other or with themselves. Furthermore, we found that myc-epitope-tagged CAP coimmunoprecipitates with HA-epitope-tagged CAP from either yeast or mammalian cell extracts. Similar results demonstrate that human CAP can also interact with human CAP2. We also show that human CAP interacts with actin, both by the yeast two-hybrid test and by coimmunoprecipitation of epitope-tagged CAP from yeast or mammalian cell extracts. This interaction requires the C-terminal domain of CAP, but not the N-terminal domain. Thus CAP appears to be capable of interacting in vivo with other CAP molecules, CAP2, and actin. We also show that actin co-immunoprecipitates with HA-CAP2 from mammalian cell extracts. © 1996 Wiley-Liss, Inc.
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  • 95
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    Effect of gene amplification on threonine production by yeast (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 49 (1996), S. 667-674 
    Details
    ISSN: 0006-3592
    Keywords: yeast ; threonine biosynthesis ; gene amplification ; amino acid production ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this work, we have studied the effect of amplifying different alleles involved in the threonine biosynthesis on the amino acid production by Saccharomyces cerevisiae. The genes used were wild-type HOM3, HOM2, HOM6, THR1, and THR4, and two mutant alleles of HOM3 (namely HOM3-R2 and HOM3-R6), that code for feedback-insensitive aspartate kinases. The results show that only the amplification of the HOM3 alleles leads to threonine and, in some instances, to homoserine overproduction. In terms of the regulation of the pathway, the data indicate that the main control is exerted by inhibition of the aspartate kinase and that, probably, a second and less important regulation takes place at the level of the homoserine kinase, the THR1 gene product. However, amplification of THR1 in two related Hom3-R2 strains does not increase the amount of threonine but, in one of them, it does induce accumulation of more homoserine. This result probably reflects differences between these strains in some undetermined genetic factor/s related with threonine metabolism. In general, the data indicate that the common laboratory yeast strains are genetically rather heterogeneous and, thus, extrapolation of conclusions must be done carefully. © 1996 John Wiley & Sons, Inc.
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  • 96
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    Efficient flotation of yeast cells grown in batch culture (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 50 (1996), S. 248-256 
    Details
    ISSN: 0006-3592
    Keywords: yeast ; Saccharomyces ; flotation ; batch culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A fast flotation assay was used to select new floating yeast strains. The flotation ability did not seem to be directly correlated to total extracellular protein concentration of the culture. However, the hydrophobicity of the cell was definitely correlated to the flotation capacity. The Saccharomyces strains (FLT strains) were highly hydrophobic and showed an excellent flotation performance in batch cultures without additives (flotation agents) and with no need for a special flotation chamber or flotation column. A stable and well-organized structure was evident in the dried foam as shown by scanning electron microscopy which revealed its unique structure showing mummified cells (dehydrated) attached to each other. The attachment among the cells and the high protein concentration of the foams indicated that proteins might be involved in the foam formation. The floating strains (strains FLT) which were not flocculent and showed no tendency to aggregate, were capable of growing and producing ethanol in a synthetic medium containing high glucose concentration as a carbon source. The phenomenon responsible for flotation seems to be quite different from the flocculation phenomenon. © 1996 John Wiley & Sons, Inc.
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  • 97
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    Control of packed column fouling in the continuous fermentation and stripping of ethanol (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 51 (1996), S. 33-39 
    Details
    ISSN: 0006-3592
    Keywords: yeast ; fuel ethanol ; flocculation ; glucose conversion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: By recycling the contents of a 14 L fermentor through a stripping column to continuously remove ethanol and reduce product inhibition, continuous complete conversion of nutrient feed containing 600 g/L glucose was achieved in a small pilot plant. Ethanol was recovered from the carbon dioxide stripping gas in a refrigerated condenser, and the gas was reheated with steam and recycled by a blower. Productivity of ethanol in the fermentor as high as 15.8 g/L/h and condensate production of up to 10 L/day of almost 50% by volume ethanol were maintained for up to 60 days of continuous operation. Weekly washing of the column packing in situ was required to prevent loss of performance caused by attached growth of yeast cells, which restricts the gas flow rate through the stripping column. © 1996 John Wiley & Sons, Inc.
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  • 98
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    Morphological characterization of the dimorphic yeast Kluyveromyces marxianus var. marxianus NRRLy2415 by semi-automated image analysis (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 51 (1996), S. 679-690 
    Details
    ISSN: 0006-3592
    Keywords: yeast ; dimorphism ; morphology ; image analysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A semiautomatic image analysis method has been developed to characterize the morphology of the dimorphic yeast Kluyveromyces marxianus var. marxianus (formerly fragilis) NRRLy2415 undergoing alcoholic fermentation of cheese whey permeate. The method is capable of separating cells into six defined categories, varying from simple ovoid yeast cells to branched mycelial cells. A sample size of 300 cells was found to be sufficient to obtain a statistically significant categorization. The processing time for a sample was found to be approximately 90 min. In addition to qualitative characterization, the method permits the measurement of geometric properties such as the width, length, and volume of individual cells or clusters of cells. When the cells analyzed by the automatic method were categorized on a manual basis, the error level in the automatic routine was found to be less than 3%.
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  • 99
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    Role of glucose signaling in yeast metabolism (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 52 (1996), S. 161-165 
    Details
    ISSN: 0006-3592
    Keywords: yeast ; signaling ; regulation ; catabolite repression ; metabolism ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this article, knowledge concerning the relation between uptake of and signaling by glucose in the yeast Saccharomyces cerevisiae is reviewed and compared to the analogous process in prokaryotes. It is concluded that (much) more fundamental knowledge concerning these processes is required before rational redesign of metabolic fluxes from glucose in yeast can be achieved. © 1996 John Wiley & Sons, Inc.
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  • 100
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    Sequencing a cosmid clone of Saccharomyces cerevisiae chromosome XIV reveals 12 new open reading frames (ORFs) and an ancient duplication of six ORFs (1996)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 391-402 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; gene duplication ; ribosomal protein ; dnaJ homologue ; fork head domain ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A sequence of 31431 bp located on the left arm of chromosome (chr.) XIV from Saccharomyces cerevisiae was analysed. A total of 18 open reading frames (ORFs) could be identified. Twelve ORFs are new, two of which are most likely ribosomal protein genes, leaving ten ORFs of unknown function. Nine of the 18 ORFs show either at least 20% overall amino acid identity or significant regional homology to other S. cerevisiae ORFs. Additionally, six of these nine ORFs have homologues of similar size and the same transcriptional orientation within a stretch of 50 kb on chromosome IX. The degree of homology ranges from 90% overall identity to 23% in 375 amino acids. The homologues on chromosome IX are grouped in two blocks that are separated by relatively long ORFs. This is the first example of a multi-gene duplication in S. cerevisiae not linked to a centromere or subtelomere region. The sequence has been deposited in the EMBL data library under Accession Number X86470.
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