Analysis of metabolic and physiological responses to gnd knockout in Escherichia coli by using C-13 tracer experiment and enzyme activity measurement

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Abstract

The physiological and metabolic responses to gnd knockout in Escherichia coli K-12 was quantitatively investigated by using the 13C 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. © 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Microbiological Societies.

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APA

Jiao, Z., Baba, T., Mori, H., & 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 Microbiology Letters, 220(2), 295–301. https://doi.org/10.1016/S0378-1097(03)00133-2

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