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Applied and Environmental Microbiology, May 2006, p. 3566-3577, Vol. 72, No. 5
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.5.3566-3577.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Short-Term Metabolome Dynamics and Carbon, Electron, and ATP Balances in Chemostat-Grown Saccharomyces cerevisiae CEN.PK 113-7D following a Glucose Pulse

Liang Wu,^1* Jan van Dam,² Dick Schipper,¹ M. T. A. Penia Kresnowati,² Angela M. Proell,² Cor Ras,² Wouter A. van Winden,² Walter M. van Gulik,² and Joseph J. Heijnen²

DSM Anti-Infectives, P.O. Box 525, NL-2613 AX Delft, The Netherlands,¹ Department of Biotechnology, Delft University of Technology, Julianalaan 67, NL-2628 BC Delft, The Netherlands²

Received 22 July 2005/ Accepted 28 February 2006

The in vivo kinetics in Saccharomyces cerevisiae CEN.PK 113-7D was evaluated during a 300-second transient period after applying a glucose pulse to an aerobic, carbon-limited chemostat culture. We quantified the responses of extracellular metabolites, intracellular intermediates in primary metabolism, intracellular free amino acids, and in vivo rates of O₂ uptake and CO₂ evolution. With these measurements, dynamic carbon, electron, and ATP balances were set up to identify major carbon, electron, and energy sinks during the postpulse period. There were three distinct metabolic phases during this time. In phase I (0 to 50 seconds after the pulse), the carbon/electron balances closed up to 85%. The accumulation of glycolytic and storage compounds accounted for 60% of the consumed glucose, caused an energy depletion, and may have led to a temporary decrease in the anabolic flux. In phase II (50 to 150 seconds), the fermentative metabolism gradually became the most important carbon/electron sink. In phase III (150 to 300 seconds), 29% of the carbon uptake was not identified in the measurements, and the ATP balance had a large surplus. These results indicate an increase in the anabolic flux, which is consistent with macroscopic balances of extracellular fluxes and the observed increase in CO₂ evolution associated with nonfermentative metabolism. The identified metabolic processes involving major carbon, electron, and energy sinks must be taken into account in in vivo kinetic models based on short-term dynamic metabolome responses.

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* Corresponding author. Mailing address: DSM Anti-Infectives, P.O. Box 525, 2613 AX Delft, The Netherlands. Phone: 31 152792513. Fax: 31 152792779. E-mail: Liang.Wu{at}dsm.com.

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Applied and Environmental Microbiology, May 2006, p. 3566-3577, Vol. 72, No. 5
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.5.3566-3577.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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