1887

Abstract

Glutamate dehydrogenase (-glutamate: NAD oxidoreductase (deaminating); EC 1.4.1.2) has been purified from in a single step using dye-ligand chromatography. The enzyme (GDH) was present in high yields and was stabilized in crude extracts. A subunit molecular weight of 49000 ±500 was determined by SDS polyacrylamide gel electrophoresis and six bands were obtained after cross-linking the subunits with dimethyl suberimidate. This bacterial GDH was predominantly NAD-linked, but was able to utilize both NADP and NADPH at 4% of the rates with NAD and NADH, respectively. An investigation of the amino acid specificity revealed some similarities with GDH from mammalian sources and some clear differences. The values of apparent for the substrates ammonia, 2-oxoglutarate, NADH, NAD and glutamate were 18·4, 0·82, 0·066, 0·031 and 6 m, respectively. The GDH was not regulated by purine nucleotides, but was subject to strong inhibition with increasing ionic strength.

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1984-09-01
2021-05-07
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