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Volume 135, Issue 12

Purification and Properties of NADP-dependent Glutamate Dehydrogenase from Free

Abstract

has two chromatographically distinct forms of glutamate dehydrogenase (GDH): one GDH utilizes NAD as coenzyme, the other uses NADP. The intracellular level of both GDHs is strongly regulated by the nitrogen source in the growth medium. NADP-dependent GDH was purified to homogeneity from crude extracts of The of the native enzyme was determined to be 200000 by size-exclusion high-performance liquid chromatography whereas after sodium dodecyl sulphate-polyacrylamide gel electrophoresis one major band of 49000 was found, suggesting that the enzyme is a tetramer. The enzyme was highly specific for the substrates 2-oxoglutarate and -glutamate, and required NADP, which could not be replaced by NAD, as a cofactor. The pH optimum was 9·2 for oxidative deamination of glutamate and 8·4 for reductive amination of 2-oxoglutarate. The Michaelis constants ( ) were 28·6 m for -glutamate and 0·12 m for NADP. values for reductive amination were 1·54 m for 2-oxoglutarate, 0·07 m for NADPH and 30·8 m for NH . The enzyme activity was significantly reduced by adenine nucleotides, particularly ATP.

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© Society for General Microbiology 1989
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1989-12-01
2025-03-22
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/content/journal/micro/10.1099/00221287-135-12-3311
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Purification and Properties of NADP-dependent Glutamate Dehydrogenase from Streptomyces fradiae
Microbiology 135, 3311 (1989); https://doi.org/10.1099/00221287-135-12-3311
/content/journal/micro/10.1099/00221287-135-12-3311
/content/journal/micro/10.1099/00221287-135-12-3311
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