The glycolytic flux in Escherichia coli is controlled by the demand for ATP.
TLDR
A molecular genetic tool is developed that specifically induces ATP hydrolysis in living cells without interfering with other aspects of metabolism, and shows that the majority of the control of growth rate resides in the anabolic reactions, i.e., the cells are mostly "carbon" limited.Abstract:
The nature of the control of glycolytic flux is one of the central, as-yet-uncharacterized issues in cellular metabolism. We developed a molecular genetic tool that specifically induces ATP hydrolysis in living cells without interfering with other aspects of metabolism. Genes encoding the F1 part of the membrane-bound (F1F0) H+-ATP synthase were expressed in steadily growing Escherichia coli cells, which lowered the intracellular [ATP]/[ADP] ratio. This resulted in a strong stimulation of the specific glycolytic flux concomitant with a smaller decrease in the growth rate of the cells. By optimizing additional ATP hydrolysis, we increased the flux through glycolysis to 1.7 times that of the wild-type flux. The results demonstrate why attempts in the past to increase the glycolytic flux through overexpression of glycolytic enzymes have been unsuccessful: the majority of flux control (>75%) resides not inside but outside the pathway, i.e., with the enzymes that hydrolyze ATP. These data further allowed us to answer the question of whether catabolic or anabolic reactions control the growth of E. coli. We show that the majority of the control of growth rate resides in the anabolic reactions, i.e., the cells are mostly “carbon” limited. Ways to increase the efficiency and productivity of industrial fermentation processes are discussed.read more
Citations
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References
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Culture Medium for Enterobacteria
TL;DR: A new minimal medium for enterobacteria is developed that supports growth of Escherichia coli and Salmonella typhimurium at rates comparable to those of any of the traditional media that have high phosphate concentrations, but each of the macronutrients is present at a sufficiently low level to permit isotopic labeling.
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Analysis of gene control signals by DNA fusion and cloning in Escherichia coli
TL;DR: Plasmid cloning vectors that enable insertion of DNA fragments between the inducible ara (arabinose) promoter and the lac (lactose) structural genes have been constructed and used for the detection and analysis of signals that control gene transcription.
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The control of flux
Henrik Kacser,J A Burns +1 more
TL;DR: Molecular Enzymology Group Colloquium Organized by D. Fell and Sponsored by Xenova Ltd, Zeneca Bioproducts, Glaxo Research and Development Ltd and SmithKline Beecham Pharmaceuticals.