title: Purification and partial characterization of the gamma-D-glutamyl-L-di-amino acid endopeptidase II from Bacillus sphaericus.
abstract: 1. A gamma-D-glutamyl-L-di-amino acid endopeptidase II (EC3.4.-.-) active on the peptide moieties of some bacterial peptidoglycans has been purified to homogeneity from the sporulation medium and from the spores of Bacillus sphaericus. 2. Enzyme from both sources showed a single protein band (Mr 28,000) by polyacrylamide gel electrophoresis under denaturing conditions. It is an acidic protein (pI 4.1). Kinetic studies have shown a Km value of 0.24 mM and an apparent Vmax of 8.3 mumol min-1 mg-1 with the pentapeptide L-Ala-gamma-D-Glu-L-Lys-D-[14C]Ala-D-[14C]Ala as substrate. 3. The enzyme was inhibited by p-hydroxymercuribenzoate, a sulfhydryl inhibitor. 4. The 38-residue N-terminal region was sequenced. It may be useful to construct a nucleotide probe for the research of the gene encoding this enzyme.
authors.affiliation: Laboratoire de Biochimie Microbienne, Université Claude Bernard Lyon I, France.
authors.forename: T
authors.initials: T
authors.lastname: Bourgogne
authors.forename: M J
authors.initials: MJ
authors.lastname: Vacheron
authors.forename: M
authors.initials: M
authors.lastname: Guinand
authors.forename: G
authors.initials: G
authors.lastname: Michel
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D004795:Enzyme Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D007526:Isoelectric Point; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D010457:Peptidoglycan / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D006902:Hydroxymercuribenzoates; D009842:Oligopeptides; D010457:Peptidoglycan; C030601:4-hydroxymercuribenzoate; D010450:Endopeptidases; C073797:gamma-D-glutamyl-L-di-amino acid endopeptidase II