PMID:12670695

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Citation

Jiao, Z, Baba, T, Mori, H and Shimizu, K (2003) Analysis of metabolic and physiological responses to gnd knockout in Escherichia coli by using C-13 tracer experiment and enzyme activity measurement. FEMS Microbiol. Lett. 220:295-301

Abstract

The physiological and metabolic responses to gnd knockout in Escherichia coli K-12 was quantitatively investigated by using the (13)C tracer experiment (GC-MS/NMR) together with the enzyme activity analysis. It was shown that the general response to the gene knockout was the local flux rerouting via Entner-Doudoroff pathway and the direction reversing via non-oxidative pentose phosphate pathway (PPP). The mutant was found to direct higher flux to phosphoglucose isomerase reaction as compared to the wild-type, but the respiratory metabolism was comparable in both strains. The anaplerotic pathway catalyzed by malic enzyme was identified in the mutant, which was accompanied with an up-regulation of phosphoenolpyruvate carboxylase and down-regulation of phosphoenolpyruvate carboxykinase. The presented results provide first evidence that compensatory mechanism existed in PPP and anaplerotic pathway in response to the gnd deletion.

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Keywords

Carbon Isotopes; Enzymes/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli/physiology; Gene Deletion; Gene Expression Regulation, Bacterial; Genes, Bacterial; Genetic Complementation Test; Models, Biological; Pentose Phosphate Pathway/physiology; Phosphogluconate Dehydrogenase/genetics; Radioactive Tracers

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