151. Flux adaptations of citrate synthase-deficient Escherichia coli
- Author
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Michael M. Domach, Akshay Goel, Mohammad M. Ataai, and J. Lee
- Subjects
biology ,General Neuroscience ,Acetyl-CoA ,Mutant ,Citric Acid Cycle ,Malic enzyme ,Citrate (si)-Synthase ,Carbon Dioxide ,Lyase ,General Biochemistry, Genetics and Molecular Biology ,Aerobiosis ,Carbon ,Citric acid cycle ,chemistry.chemical_compound ,Adenosine Triphosphate ,History and Philosophy of Science ,chemistry ,Biochemistry ,Bacterial Proteins ,biology.protein ,Escherichia coli ,Citrate synthase ,Formate ,Flux (metabolism) ,Glycolysis - Abstract
The results from the experiments performed with a mutant deficient in citrate synthase activity can be summarized as follows. (1) Totally blocking entry into the TCA cycle did not appreciably alter the cellular ATP yield. The unchanged yield suggests that for growth on abundant glucose, the sensitivity of ATP yield to TCA cycle flux is low. ATP production in the mutant is altered, in part, by modulating the relative amounts of formate and acetate produced. (2) The in vivo operation of pyruvate-formate lyase and malic enzyme corresponds to proposals developed from in vitro studies. Namely, pyruvate activates the former, and acetyl CoA inhibits the latter. Overall, the diversion of pyruvate to formate under aerobic conditions constitutes an adaptation of the mutant to the enzymatic lesion. The low alpha-ketoglutarate dehydrogenase flux estimated for the mutant indicates that the enzyme is highly repressed in cells growing rapidly on glucose, which is in accord with prior induction-repression studies. Moreover, the lack of a change in uptake flux during the bulk of batch growth is consistent with prior induction-repression studies. (3) The mutant exhibits a heightened sensitivity to CO2 as compared to wild-type counterparts. Growth rate is increased, and the production of formate, malate, glycerate, and pyruvate is reduced. This sensitivity illustrates that citrate synthase is more than an expendable component in an amphibolic pathway. Its presence in wild-type cells "immunizes" against the effect of CO2 fluctuations. (4) The effects of CO2 can be tentatively explained by assuming that the PEP carboxylase-catalyzed reaction is stimulated.
- Published
- 1994