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Strategies to determine the extent of control exerted by glucose transport on glycolytic flux in the yeast Saccharomyces bayanus

E. C. Slater Institute, BioCentrum, University of Amsterdam, Plantage Muidergracht 12, NL-1018 TV Amsterdam, The Netherlands¹
Biotechnology Unit, Faculty of Sciences and Technology, New University of Lisbon, P-2825 Monte de Caparica, Portugal ²

Author for correspondence: Karel van Dam. Tel: +31 20 525 5510. Fax: +31 20 525 5505. e-mail: k.van.dam{at}chem.uva.nl

The extent to which the transport of glucose across the plasma membrane of the yeast Saccharomyces bayanus controls the glycolytic flux was determined. The magnitude of control was quantified by measuring the effect of small changes in the activity of the glucose transport system on the rate of glucose consumption. Two effectors were used to modulate the activity of glucose transport: (i) maltose, a competitive inhibitor of the glucose transport system in S. bayanus (as well as in Saccharomyces cerevisiae) and (ii) extracellular glucose, the substrate of the glucose transport system. Two approaches were followed to derive from the experimental data the flux control coefficient of glucose transport on the glycolytic flux: (i) direct comparison of the steady-state glycolytic flux with the zero trans-influx of glucose and (ii) comparison of the change in glycolytic flux with the concomitant change in calculated glucose transport activity on variation of the extracellular glucose concentration. Both these approaches demonstrated that in cells of S. bayanus grown on glucose and harvested at the point of glucose exhaustion, a high proportion of the control of the glycolytic flux resides in the transport of glucose across the plasma membrane.

Keywords: yeast (Saccharomyces), glucose transport, maltose inhibition, glycolysis, metabolic control analysis (MCA)

Abbreviations: MCA, metabolic control analysis

^a Present address: Instituut voor Interdisciplinaire Opleidingen (I2O), Sarphatistraat 104, NL-1018 GV Amsterdam, The Netherlands.

^b Present address: Heineken Technical Services, Burgemeester Smeetsweg 1, NL-2382 PH Zoeterwoude, The Netherlands.

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