Activities of the enzymes of the Ehrlich pathway and formation of branched-chain alcohols in Saccharomyces cerevisiae and Candida utilis grown in continuous culture on valine or ammonium as sole nitrogen source

Derrick, Simon; Large, Peter J.

doi:10.1099/00221287-139-11-2783

Cited by 84 publications

(54 citation statements)

References 44 publications

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“…As expected, the LEU2 gene showed only background expression, while genes coding for branched-chain amino acid aminotransferases, i.e., BAT1 and BAT2, showed inverse expression correlating with the cells transitioning from logarithmic to growth-arrested phase (10). It is therefore likely that the improved growth of the SPT15-300 mutant under leucine-limiting conditions is due to increased uptake and utilization of leucine (9,28,36).…”

Section: Resultsmentioning

confidence: 89%

Impaired Uptake and/or Utilization of Leucine bySaccharomyces cerevisiaeIs Suppressed by theSPT15-300Allele of the TATA-Binding Protein Gene

Baerends

Qiu

Rasmussen

et al. 2009

Appl Environ Microbiol

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Improvement of stress tolerance in microorganisms applied in industrial fermentations for the production of ethanol is of major interest (26, 34). Based on screens for ethanol sensitivity/tolerance in Saccharomyces cerevisiae (12, 16-18, 35, 37, 40), it appears that this trait in yeast is possibly controlled by several genes acting in concert. Using global transcription machinery engineering (gTME), a tool to reprogram gene transcription for eliciting new phenotypes important for technological applications, Alper et al. (2) found mutants of S. cerevisiae with improved glucose/ethanol tolerance. In that work, mutated versions of the SPT15 gene, which codes for the TATA-binding protein, were generated by random in vitro mutagenesis and expressed in the laboratory strain BY4741. The authors identified one dominant allele, SPT15-300, which corresponds to the three amino acid changes F177S, Y195H, and K218R, that conferred increased tolerance of the yeast to ethanol (2). Although extensive analyses, such as transcriptional profiling and deleting and overexpressing individual genes, were carried out, a particular pathway or a genetic network responsible for the observed growth gain of the SPT15-300-expressing strain could not be identified (2). During our attempts to analyze the effect of the mutant SPT15-300 alleles in various yeast species of industrial importance, we discovered that the described improvement of growth in the presence of ethanol of the standard laboratory strain BY4741, the strain used by Alper et al. (2), is associated with improved uptake and/or utilization of leucine on media containing small amounts of leucine. MATERIALS AND METHODSStrains, media, and molecular procedures. The Saccharomyces strains investigated in this study were S. cerevisiae strains BY4741 (MATa his3⌬D1 leu2⌬0 met15⌬0 ura3⌬0) (5), in which the LEU2 gene is completely deleted (obtained from Euroscarf, Frankfurt, Germany), and Y55 (23) 2), and YSC lacking leucine (prepared as described for YSCϪUra, with Qbiogene CSM-LEU [2]). SC media were buffered (pH 5.5) with 1% (wt/vol) succinic acid and 0.6% (wt/vol) NaOH. SC and YSC media were supplemented with glucose and/or ethanol as indicated. S. cerevisiae strains were incubated at 20 or 30°C (as indicated), and S. bayanus and S. pastorianus were cultivated at 20°C. Saccharomyces species were transformed by use of the lithium acetate method (3).Escherichia coli strain DH5␣ (Invitrogen A/S, Taastrup, Denmark) was used for plasmid selection/propagation and cultivated as described previously (31).

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Section: Resultsmentioning

confidence: 89%

Impaired Uptake and/or Utilization of Leucine bySaccharomyces cerevisiaeIs Suppressed by theSPT15-300Allele of the TATA-Binding Protein Gene

Baerends

Qiu

Rasmussen

et al. 2009

Appl Environ Microbiol

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“…The irreversible decarboxylation step, which commits 2-oxo acids to the Ehrlich pathway, was initially attributed to pyruvate decarboxylase (5,55,65). However, the yeast genome was shown to harbor no fewer than five genes that show sequence similarity with thiamine diphosphate (TPP)-dependent decarboxylase genes.…”

Section: Decarboxylation Stepmentioning

confidence: 99%

The Ehrlich Pathway for Fusel Alcohol Production: a Century of Research on Saccharomyces cerevisiae Metabolism

Hazelwood

Daran

Maris

et al. 2008

Appl Environ Microbiol

1,210

613

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“…Thus, we suggest that ARO9 should be added to the list of Rim101p-regulated genes. Dual regulation of ARO9 by Aro80p and Rim101p suggests that Aro9p is a critical step for the regulation of fusel oils, a reasonable possibility because the decarboxylation step that follows is effectively irreversible (12,18,34). For ARO10, regulation was lost in rim101 but not in rim13 (Fig.…”

Section: Discussionmentioning

confidence: 98%

Regulation of Aromatic Alcohol Production in Candida albicans

Ghosh

Kebaara

Atkin

et al. 2008

Appl Environ Microbiol

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Colonization by the fungal pathogen Candida albicans varies significantly, depending upon the pH and availability of oxygen. Because of our interest in extracellular molecules as potential quorum-sensing molecules, we examined the physiological conditions which regulate the production of the aromatic alcohols, i.e., phenethyl alcohol, tyrosol, and tryptophol. The production of these fusel oils has been well studied for Saccharomyces cerevisiae. Our data show that aromatic alcohol yields for C. albicans are determined by growth conditions. These conditions include the availability of aromatic amino acids, the pH, oxygen levels, and the presence of ammonium salts. For example, for wild-type C. albicans, tyrosol production varied 16-fold merely with the inclusion of tyrosine or ammonium salts in the growth medium. Aromatic alcohol production also depends on the transcription regulator Aro80p. Our results are consistent with aromatic alcohol productionaromatic transaminases (gene products for ARO8 and ARO9), aromatic decarboxylase (ARO10), and alcohol dehydrogenase (ADH)-via the fusel oil pathway. The expression of ARO8, ARO9, and ARO10 is also pH dependent. ARO8 and ARO9 were alkaline upregulated, while ARO10 was alkaline downregulated. The alkalinedependent change in expression of ARO8 was Rim101 independent, while the expression of ARO9 was Rim101 dependent.

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Activities of the enzymes of the Ehrlich pathway and formation of branched-chain alcohols in Saccharomyces cerevisiae and Candida utilis grown in continuous culture on valine or ammonium as sole nitrogen source

Cited by 84 publications

References 44 publications

Impaired Uptake and/or Utilization of Leucine bySaccharomyces cerevisiaeIs Suppressed by theSPT15-300Allele of the TATA-Binding Protein Gene

Impaired Uptake and/or Utilization of Leucine bySaccharomyces cerevisiaeIs Suppressed by theSPT15-300Allele of the TATA-Binding Protein Gene

The Ehrlich Pathway for Fusel Alcohol Production: a Century of Research on Saccharomyces cerevisiae Metabolism

Regulation of Aromatic Alcohol Production in Candida albicans

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