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Published March 1994 | metadata_only
Journal Article

Ethanol production in baker's yeast: Application of experimental perturbation techniques for model development and resultant changes in flux control analysis


Recent theoretical results (Schlosser and Bailey, 1990; Schlosser et al., 1993) suggest that it is possible to model elements of metabolism to the extent necessary for flux control analysis using only the results of perturbation experiments. In particular, the development of detailed, mechanistic descriptions of certain relevant processes (which would otherwise require prohibitive effort) can be avoided by direct use of experimental results. Anaerobic glycolysis in Saccharomyces cerevisiae AMW-13C was studied both under unperturbed conditions and under several experimental perturbations. Changes in pathway fluxes and metabolite levels relative to the unperturbed values were observed and analyzed to determine the relationships that exist between several of these quantities. These relationships were used to refine and extend a previously determined model for the glycolytic pathway (Galazzo and Bailey, 1990,1991). The refined model was then used to study the flux control characteristics of the pathway. The analysis indicates that the reaction catalyzed by phosphofructokinase (PFK), while retaining significant flux control, is less important than previous studies suggested and that glucose uptake is the predominant rate-controlling step when small changes are considered. The model indicates that larger increases in the amount of any single pathway enzyme yield no more than a 36% increase in ethanol production.

Additional Information

© 1994 American Chemical Society. Accepted November 4, 1993. The authors thank Dr. Susan Sumner at the Chemical Industry Institute of Toxicology (CIIT) for her very helpful comments on the NMR analysis. We are deeply indebted to Ed Bailey (no relation to J.E.B.) at RhGne-Poulenc AG Co. for his assistance in preparing the NMR spectra for publication. Sincere thanks go to Dr. Derek Janszen at CIIT for his assistance with the statistical analysis. The authors thank Dr. J. Heinisch for providing Saccharomyces cerevisiae strain AMW-13C. This work was supported by the National Science Foundation (Grant No. BCS 8912824). P.S. was supported by the Department of Health and Human Services, National Research Service Award No. 1 F32 GM 13622-01 BIOM, from the National Institute of General Medical Sciences.

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August 20, 2023
August 20, 2023