Metabolic-flux analysis of Saccharomyces cerevisiae CEN.PK113-7D based on mass isotopomer measurements of 13C-labeled primary metabolites

Wouter A. Van Winden*, Jan C. Van Dam, Cor Ras, Roelco J. Kleijn, Jacobus L. Vinke, Walter M. Van Gulik, Joseph J. Heijnen

*Corresponding author for this work

    Research output: Contribution to journalArticleScientificpeer-review

    140 Citations (Scopus)


    Metabolic-flux analyses in microorganisms are increasingly based on 13C-labeling data. In this paper a new approach for the measurement of 13C-label distributions is presented: rapid sampling and quenching of microorganisms from a cultivation, followed by extraction and detection by liquid chromatography-mass spectrometry of free intracellular metabolites. This approach allows the direct assessment of mass isotopomer distributions of primary metabolites. The method is applied to the glycolytic and pentose phosphate pathways of Saccharomyces cerevisiae strain CEN.PK113-7D grown in an aerobic, glucose-limited chemostat culture. Detailed investigations of the measured mass isotopomer distributions demonstrate the accuracy and information-richness of the obtained data. The mass fractions are fitted with a cumomer model to yield the metabolic fluxes. It is estimated that 24% of the consumed glucose is catabolized via the pentose phosphate pathway. Furthermore, it is found that turnover of storage carbohydrates occurs. Inclusion of this turnover in the model leads to a large confidence interval of the estimated split ratio.

    Original languageEnglish
    Pages (from-to)559-568
    Number of pages10
    JournalFEMS Yeast Research
    Issue number6-7
    Publication statusPublished - 2005


    • C-labeling
    • Glycolysis
    • LC-MS/MS
    • Metabolic-flux analysis
    • Pentose phosphate pathway
    • Saccharomyces cerevisiae


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