Metabolic flux screening of Saccharomyces cerevisiae single knockout strains on glucose and galactose supports elucidation of gene function

Vidya R. Velagapudi, Christoph Wittmann, Konstantin Schneider, Elmar Heinzle

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

New methods for an extended physiological characterization of yeast at a microtiter plate scale were applied to 27 deletion mutants of Saccharomyces cerevisiae cultivated on glucose and galactose as sole carbon sources. In this way, specific growth rates, specific rates of glucose consumption and ethanol production were determined. Flux distribution, particularly concerning branching into the pentose phosphate pathway was determined using a new 13C-labelling method using MALDI-ToF-mass spectrometry. On glucose, the growth was predominantly fermentative whereas on galactose respiration was more active with correspondingly lower ethanol production. Some deletion strains showed unexpected behavior providing very informative data about the function of the corresponding gene. Deletion of malic enzyme gene, MAE1, did not show any significant phenotype when grown on glucose but a drastically increased branching from glucose 6-phosphate into the pentose phosphate pathway when grown on galactose. This allows the conclusion that MAE1 is important for the supply of NADPH during aerobic growth on galactose.
Original languageEnglish
JournalJournal of Biotechnology
Volume132
Issue number4
Pages (from-to)395-404
Number of pages10
ISSN0168-1656
DOIs
Publication statusPublished - 2007
Externally publishedYes

Keywords

  • Saccharomyces cerevisiae
  • Malic enzyme
  • Microtiter plate cultivation
  • MALDI-ToF-mass spectrometry
  • Phenotypic analysis

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