Characterization of GCY1 in Saccharomyces cerevisiae by metabolic profiling

J. Y. Jung, T. Y. Kim, C. Y. Ng, M. K. Oh

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Aims: The analytical study of intracellular (IC) metabolites has developed with advances in chromatography-linked mass spectrometry and fast sampling procedures. We applied the IC metabolite analysis to characterize the role of GCY1 in the glycerol (GLY) catabolic pathway in Saccharomyces cerevisiae. Methods and Results: Strains with disrupted or overexpressing GLY catabolic genes such as GCY1, DAK1 and DAK2 were constructed. The strains were cultivated under different aeration conditions and quickly quenched using a novel rapid sampling port. IC concentrations of GLY, dihydroxyacetone (DHA), glycerol 3-phosphate and dihydroxyacetone phosphate were analysed in the strains by gas chromatography/mass spectrometry. DHA was not detected in the gcy1 gene-disrupted strain but accumulated 225·91 μmol g DCW-1 in a DHA kinase gene-deficient strain under micro-aerobic conditions. Additionally, a 16·1% increase in DHA occurred by overexpressing GCY1 in the DHA kinase-deficient strain. Conclusions: Metabolic profiling showed that the GCY1 gene product functions as a GLY dehydrogenase in S. cerevisiae, particularly under micro-aerobic conditions. Significance and Impact of the Study: Metabolic profiling of the GLY dissimilation pathway was successfully demonstrated in S. cerevisiae, and the function of GCY1 was explained by the results.

Original languageEnglish
Pages (from-to)1468-1478
Number of pages11
JournalJournal of Applied Microbiology
Volume113
Issue number6
DOIs
Publication statusPublished - 2012 Dec

Keywords

  • Fast sampling device
  • GCY1
  • Glycerol
  • Metabolomics
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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