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Dynamics of intracellular metabolites of glycolysis and TCA cycle during cell-cycle-related oscillation in Saccharomyces cerevisiae.
Biotechnol Bioeng. 2005 Mar 30; 89(7):839-47.BB

Abstract

In the present work LC-MS/MS was applied to measure the concentrations of intermediates of glycolysis and TCA cycle during autonomous, cell-cycle synchronized oscillations in aerobic, glucose-limited chemostat cultures of Saccharomyces cerevisiae. This study complements previously reported oscillations in carbon dioxide production rate, intracellular concentrations of trehalose and various free amino acids, and extracellular acetate and pyruvate in the same culture. Of the glycolytic intermediates, fructose 1,6-bisphosphate, 2- and 3-phosphoglycerate, and phosphoenolpyruvate show the most pronounced oscillatory behavior, the latter three compounds oscillating out of phase with the former. This agrees with previously observed metabolic control by phosphofructokinase and pyruvate kinase. Although individually not clearly oscillating, several intermediates of the TCA cycle, i.e., alpha-ketoglutarate, succinate, fumarate, and malate, exhibited increasing concentration during the cell cycle phase with high carbon flux through glycolysis and TCA cycle. The average mass action ratios of beta-phosphoglucomutase and fumarase agreed well with previously determined in vitro equilibrium constants. Minor differences resulted for phosphoglucose isomerase and enolase. Together with the observed close correlation of the pool sizes of the involved metabolites, this might indicate that, in vivo, these reactions are operating close to equilibrium, whereby care must be taken due to possible differences between in vivo and in vitro conditions. Combining the data with previously determined intracellular amino acid levels from the same culture, a few clear correlations between catabolism and anabolism could be identified: phosphoglycerate/serine and alpha-ketoglutarate/lysine exhibited correlated oscillatory behavior, albeit with different phase shifts. Oscillations in intracellular amino acids might therefore be, at least partly, following oscillations of their anabolic precursors.

Authors+Show Affiliations

Biochemical Engineering, Saarland University, POB 151150, 66123 Saarbrücken, Germany. c.wittmann@mx.uni-saarland.deNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

15690349

Citation

Wittmann, Christoph, et al. "Dynamics of Intracellular Metabolites of Glycolysis and TCA Cycle During Cell-cycle-related Oscillation in Saccharomyces Cerevisiae." Biotechnology and Bioengineering, vol. 89, no. 7, 2005, pp. 839-47.
Wittmann C, Hans M, van Winden WA, et al. Dynamics of intracellular metabolites of glycolysis and TCA cycle during cell-cycle-related oscillation in Saccharomyces cerevisiae. Biotechnol Bioeng. 2005;89(7):839-47.
Wittmann, C., Hans, M., van Winden, W. A., Ras, C., & Heijnen, J. J. (2005). Dynamics of intracellular metabolites of glycolysis and TCA cycle during cell-cycle-related oscillation in Saccharomyces cerevisiae. Biotechnology and Bioengineering, 89(7), 839-47.
Wittmann C, et al. Dynamics of Intracellular Metabolites of Glycolysis and TCA Cycle During Cell-cycle-related Oscillation in Saccharomyces Cerevisiae. Biotechnol Bioeng. 2005 Mar 30;89(7):839-47. PubMed PMID: 15690349.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Dynamics of intracellular metabolites of glycolysis and TCA cycle during cell-cycle-related oscillation in Saccharomyces cerevisiae. AU - Wittmann,Christoph, AU - Hans,Michael, AU - van Winden,Wouter A, AU - Ras,Cor, AU - Heijnen,Joseph J, PY - 2005/2/4/pubmed PY - 2005/6/15/medline PY - 2005/2/4/entrez SP - 839 EP - 47 JF - Biotechnology and bioengineering JO - Biotechnol Bioeng VL - 89 IS - 7 N2 - In the present work LC-MS/MS was applied to measure the concentrations of intermediates of glycolysis and TCA cycle during autonomous, cell-cycle synchronized oscillations in aerobic, glucose-limited chemostat cultures of Saccharomyces cerevisiae. This study complements previously reported oscillations in carbon dioxide production rate, intracellular concentrations of trehalose and various free amino acids, and extracellular acetate and pyruvate in the same culture. Of the glycolytic intermediates, fructose 1,6-bisphosphate, 2- and 3-phosphoglycerate, and phosphoenolpyruvate show the most pronounced oscillatory behavior, the latter three compounds oscillating out of phase with the former. This agrees with previously observed metabolic control by phosphofructokinase and pyruvate kinase. Although individually not clearly oscillating, several intermediates of the TCA cycle, i.e., alpha-ketoglutarate, succinate, fumarate, and malate, exhibited increasing concentration during the cell cycle phase with high carbon flux through glycolysis and TCA cycle. The average mass action ratios of beta-phosphoglucomutase and fumarase agreed well with previously determined in vitro equilibrium constants. Minor differences resulted for phosphoglucose isomerase and enolase. Together with the observed close correlation of the pool sizes of the involved metabolites, this might indicate that, in vivo, these reactions are operating close to equilibrium, whereby care must be taken due to possible differences between in vivo and in vitro conditions. Combining the data with previously determined intracellular amino acid levels from the same culture, a few clear correlations between catabolism and anabolism could be identified: phosphoglycerate/serine and alpha-ketoglutarate/lysine exhibited correlated oscillatory behavior, albeit with different phase shifts. Oscillations in intracellular amino acids might therefore be, at least partly, following oscillations of their anabolic precursors. SN - 0006-3592 UR - https://www.unboundmedicine.com/medline/citation/15690349/Dynamics_of_intracellular_metabolites_of_glycolysis_and_TCA_cycle_during_cell_cycle_related_oscillation_in_Saccharomyces_cerevisiae_ DB - PRIME DP - Unbound Medicine ER -