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Applied and Environmental Microbiology, September 2008, p. 5710-5723, Vol. 74, No. 18
0099-2240/08/$08.00+0     doi:10.1128/AEM.01121-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Dynamics of Glycolytic Regulation during Adaptation of Saccharomyces cerevisiae to Fermentative Metabolism{triangledown} ,{dagger}

Joost van den Brink, André B. Canelas, Walter M. van Gulik, Jack T. Pronk, Joseph J. Heijnen, Johannes H. de Winde, and Pascale Daran-Lapujade*

Kluyver Centre for Genomics of Industrial Fermentation and Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands

Received 19 May 2008/ Accepted 14 July 2008

The ability of baker's yeast (Saccharomyces cerevisiae) to rapidly increase its glycolytic flux upon a switch from respiratory to fermentative sugar metabolism is an important characteristic for many of its multiple industrial applications. An increased glycolytic flux can be achieved by an increase in the glycolytic enzyme capacities (Vmax) and/or by changes in the concentrations of low-molecular-weight substrates, products, and effectors. The goal of the present study was to understand the time-dependent, multilevel regulation of glycolytic enzymes during a switch from fully respiratory conditions to fully fermentative conditions. The switch from glucose-limited aerobic chemostat growth to full anaerobiosis and glucose excess resulted in rapid acceleration of fermentative metabolism. Although the capacities (Vmax) of the glycolytic enzymes did not change until 45 min after the switch, the intracellular levels of several substrates, products, and effectors involved in the regulation of glycolysis did change substantially during the initial 45 min (e.g., there was a buildup of the phosphofructokinase activator fructose-2,6-bisphosphate). This study revealed two distinct phases in the upregulation of glycolysis upon a switch to fermentative conditions: (i) an initial phase, in which regulation occurs completely through changes in metabolite levels; and (ii) a second phase, in which regulation is achieved through a combination of changes in Vmax and metabolite concentrations. This multilevel regulation study qualitatively explains the increase in flux through the glycolytic enzymes upon a switch of S. cerevisiae to fermentative conditions and provides a better understanding of the roles of different regulatory mechanisms that influence the dynamics of yeast glycolysis.

* Corresponding author. Mailing address: Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands. Phone: 31 15 278 99 65. Fax: 31 15 278 23 55. E-mail: p.a.s.daran-lapujade{at}tudelft.nl

{triangledown} Published ahead of print on 18 July 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, September 2008, p. 5710-5723, Vol. 74, No. 18
0099-2240/08/$08.00+0     doi:10.1128/AEM.01121-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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