The gene for Escherichia coli glutamate dehydrogenase (EcGDH) has been overexpressed, and a simplified purification procedure afforded greatly increased yields of c. 40 mg pure EcGDH L-1 culture. EcGDH was unstable at a low concentration in plastic tubes, but stabilization measures allowed a robust kinetic characterization. Contrary to past reports, EcGDH deviates from Michaelis-Menten kinetics, exhibiting apparent mild negative co-operativity with both l-glutamate and NADP+, with Hill coefficients of 0.90 and 0.92, respectively. NADPH yielded simple Michaelis-Menten kinetics but both 2-oxoglutarate and NH4+ showed substrate inhibition. pH optima were 9 for oxidative deamination and 8 for reductive amination. © 2008 Federation of European Microbiological Societies Published by Blackwell Publishing Ltd.
CITATION STYLE
Sharkey, M. A., & Engel, P. C. (2008). Apparent negative co-operativity and substrate inhibition in overexpressed glutamate dehydrogenase from Escherichia coli. FEMS Microbiology Letters, 281(2), 132–139. https://doi.org/10.1111/j.1574-6968.2008.01086.x
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