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Characterization of GCY1 in S accharomyces cerevisiae by metabolic profiling
Author(s) -
Jung J.Y.,
Kim T.Y.,
Ng C.Y.,
Oh M.K.
Publication year - 2012
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12013
Subject(s) - saccharomyces cerevisiae , dihydroxyacetone , dihydroxyacetone phosphate , biochemistry , catabolism , metabolite , biology , metabolome , glycerol kinase , glycerol , dehydrogenase , metabolism , gene , chemistry , enzyme
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 S accharomyces 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.