title: Isolation and characterization of glutamyl endopeptidase 2 from Bacillus intermedius 3-19.
abstract: The culture filtrate of Bacillus intermedius 3-19 was used for isolation by chromatography on CM-cellulose and Mono S columns of a proteinase that is secreted during the late stages of growth. The enzyme is irreversibly inhibited by the inhibitor of serine proteinases diisopropyl fluorophosphate, has two pH optima (7.2 and 9.5) for casein hydrolysis and one at pH 8.5 for Z-Glu-pNA hydrolysis. The molecular weight of the enzyme is 26.5 kD. The K(m) for Z-Glu-pNA hydrolysis is 0.5 mM. The temperature and pH dependences of the stability of the proteinase were studied. The enzyme was identified as glutamyl endopeptidase 2. The N-terminal sequence (10 residues) and amino acid composition of the enzyme were determined. The enzyme hydrolyzes Glu4-Gln5, Glu17-Asp18, and Cys11-Ser12 bonds in the oxidized A-chain of insulin and Glu13-Ala14, Glu21-Arg22, Cys7-Gly8, and Cys19-Gly20 bonds in the oxidized B-chain of insulin.
authors.affiliation: Faculty of Biology, Kazan State University, Kazan, 420008, Russia. Nelly.Balaban@ksu.ru
authors.forename: N P
authors.initials: NP
authors.lastname: Balaban
authors.forename: A M
authors.initials: AM
authors.lastname: Mardanova
authors.forename: M R
authors.initials: MR
authors.lastname: Sharipova
authors.forename: L A
authors.initials: LA
authors.lastname: Gabdrakhmanova
authors.forename: E A
authors.initials: EA
authors.lastname: Sokolova
authors.forename: A V
authors.initials: AV
authors.lastname: Garusov
authors.forename: E I
authors.initials: EI
authors.lastname: Milgotina
authors.forename: G N
authors.initials: GN
authors.lastname: Rudenskaya
authors.forename: I B
authors.initials: IB
authors.lastname: Leshchinskaya
mesh_terms: D001407:Bacillus / Q000201:enzymology* / Q000254:growth & development; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D011480:Protease Inhibitors / Q000494:pharmacology; D020539:Sequence Analysis, Protein; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification; D013379:Substrate Specificity / Q000187:drug effects
chemical_list: D011480:Protease Inhibitors; D012697:Serine Endopeptidases