Contributions to the kinetic modeling of glycolytic pathway in yeast

Şahin, Ceylan
Being at the center of most metabolic pathways and also one of the best known pathways, the glycolytic pathway has been of interest to modeling studies. This study is composed of our attempts to model ethanolic fermentation by yeast through kinetic equations of glycolytic steps and its branches. Model was based totally on experimentally measured kinetics of enzymes and transport steps, either obtained in this study or from the literature. Effect of ethanol on enzyme activities was tested in the range of ethanol 0 to 20% (v/v) in assay mixture. All enzymes were inhibited by ethanol to some degree and these inhibitions started at different ethanol concentrations, the least affected being the pyruvate kinase and the most inhibited ones being glycerol-3-phosphate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, phosphogluco kinase, and alcohol dehydrogenase (forward). Effect of temperature on the activities of enzymes was tested within 10-30 °C with five degrees of increments. Activation energies of enzymes were calculated using the Arrhenius equation. Activation energies of upper part of the glycolysis and the glycerol branch (glycerol-3-phosphate dehydrogenase) were relatively higher than that of lower part enzymes as well as the ethanol branch (alcohol dehydrogenase). Results obtained from these in vitro studies were incorporated into the model as mathematical relations. Model output thus obtained was compared with results of experiments conducted at several temperatures and initial ethanol concentrations. Model could estimate general trend in ethanolic fermentation that fermentation is inhibited by increasing concentrations of ethanol. Decrease in glycerol yields at lower temperatures was also estimated by the model. However, model did not fit exactly to experimental results, especially at low temperature and high ethanol concentrations. This could be attributed to stress responses of cells under these conditions, which are not considered in the model.


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Citation Formats
C. Şahin, “Contributions to the kinetic modeling of glycolytic pathway in yeast,” Ph.D. - Doctoral Program, Middle East Technical University, 2009.