title: Ammonia assimilation in Klebsiella pneumoniae F-5-2 that can utilize ammonium and nitrate ions simultaneously: purification and characterization of glutamate dehydrogenase and glutamine synthetase.
abstract: The two ammonia-assimilating enzymes glutamate dehydrogenase (GDH; EC 1.4.1.4) and glutamine synthetase (GS; EC 6.3.1.2) were synthesized steadily during the cell growth of Klebsiella pneumoniae F-5-2 that can utilize NH4+ and NO3- simultaneously under aerobic conditions. The enzymes were purified to homogeneity from cell extracts and characterized. The molecular mass of the purified GDH was 300 kDa with six identical 52-kDa subunits. GDH showed its maximal activity (aminating) at pH 8.0 and was stable between pHs 5.5 and 11.5. The enzyme was NADP-specific and strongly inhibited by Ag+. It catalyzed the amination of 2-ketovalerate, 2-ketoadipate, and 2-ketobutyrate, in addition to 2-ketoglutarate. The purified GS has a molecular mass of 470 kDa with eight identical 60-kDa subunits. GS showed its maximal activity at pH 8.0 and was stable between pHs 6.0 and 7.0. The enzyme was strongly inhibited by Fe3+, Hg2+, and Cu2+.
authors.affiliation: Division ofLife Science, The Graduate School ofScience and Technology, Kobe University, Rokko, Kobe 657-8501, Japan.
authors.forename: Young-Ju
authors.initials: YJ
authors.lastname: Kim
authors.forename: Makoto
authors.initials: M
authors.lastname: Yoshizawa
authors.forename: Shinji
authors.initials: S
authors.lastname: Takenaka
authors.forename: Shuichiro
authors.initials: S
authors.lastname: Murakami
authors.forename: Kenji
authors.initials: K
authors.lastname: Aoki