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Journal of Bacteriology, February 2001, p. 1441-1451, Vol. 183, No. 4
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.4.1441-1451.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Network Identification and Flux Quantification in the Central Metabolism of Saccharomyces cerevisiae under Different Conditions of Glucose Repression

Andreas Karoly Gombert,dagger Margarida Moreira dos Santos, Bjarke Christensen, and Jens Nielsen*

Center for Process Biotechnology, Department of Biotechnology, Technical University of Denmark, DK-2800, Lyngby, Denmark

Received 22 June 2000/Accepted 23 November 2000

The network structure and the metabolic fluxes in central carbon metabolism were characterized in aerobically grown cells of Saccharomyces cerevisiae. The cells were grown under both high and low glucose concentrations, i.e., either in a chemostat at steady state with a specific growth rate of 0.1 h-1 or in a batch culture with a specific growth rate of 0.37 h-1. Experiments were carried out using [1-13C]glucose as the limiting substrate, and the resulting summed fractional labelings of intracellular metabolites were measured by gas chromatography coupled to mass spectrometry. The data were used as inputs to a flux estimation routine that involved appropriate mathematical modelling of the central carbon metabolism of S. cerevisiae. The results showed that the analysis is very robust, and it was possible to quantify the fluxes in the central carbon metabolism under both growth conditions. In the batch culture, 16.2 of every 100 molecules of glucose consumed by the cells entered the pentose-phosphate pathway, whereas the same relative flux was 44.2 per 100 molecules in the chemostat. The tricarboxylic acid cycle does not operate as a cycle in batch-growing cells, in contrast to the chemostat condition. Quantitative evidence was also found for threonine aldolase and malic enzyme activities, in accordance with published data. Disruption of the MIG1 gene did not cause changes in the metabolic network structure or in the flux pattern.

* Corresponding author. Mailing address: Center for Process Biotechnology, Department of Biotechnology, Technical University of Denmark, Building 223, DK-2800, Lyngby, Denmark. Phone: 45 45 25 2696. Fax: 45 45 88 4148. E-mail: jn{at}ibt.dtu.dk.

dagger Present address: Department of Chemical Engineering, University of São Paulo, São Paulo, Brazil.

Journal of Bacteriology, February 2001, p. 1441-1451, Vol. 183, No. 4
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.4.1441-1451.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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