RT Journal Article SR Electronic(1) A1 Hornby, David P. A1 Engel, Paul C.YR 1984 T1 Characterization of Peptostreptococcus asaccharolyticus Glutamate Dehydrogenase Purified by Dye-ligand Chromatography JF Microbiology, VO 130 IS 9 SP 2385 OP 2394 DO https://doi.org/10.1099/00221287-130-9-2385 PB Microbiology Society, SN 1465-2080, AB Glutamate dehydrogenase (l-glutamate: NAD+ oxidoreductase (deaminating); EC 1.4.1.2) has been purified from Peptostreptococcus asaccharolyticus 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 K m for the substrates ammonia, 2-oxoglutarate, NADH, NAD+ and glutamate were 18·4, 0·82, 0·066, 0·031 and 6 mm, respectively. The P. asaccharolyticus GDH was not regulated by purine nucleotides, but was subject to strong inhibition with increasing ionic strength., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-130-9-2385