title: Purification and characterization of a major collagenase from Streptomyces parvulus.
abstract: A major collagenase was purified about 96-fold from a crude enzyme sample of Streptomyces parvulus by chromatography on Q-Sepharose, Sephacryl S-200, and butyl-Toyopearl. The purified enzyme showed a relative molecular mass of approximately 52,000 on SDS-PAGE and a pH optimum at about 9.0, and was strongly inhibited by metal-chelating agents. It also cleaved 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg specifically at the Leu-Gly bond, with a K(m) value of 0.60 mM at pH 9.0 at 37 degrees C. Based on the amino acid sequences of the N-terminal region and internal tryptic peptides, the corresponding gene was cloned. The DNA sequence of the cloned gene indicated that the enzyme is produced as an 864-residue precursor protein with a 408-residue prepro sequence followed by a 456-residue mature enzyme moiety. The enzyme is most homologous with the collagenase from S. coelicolor, the identity being 73%, and it is thought to be a member of the Vibrio collagenase subfamily.
authors.affiliation: Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo.
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Sakurai
authors.forename: Hideshi
authors.initials: H
authors.lastname: Inoue
authors.forename: Wataru
authors.initials: W
authors.lastname: Nishii
authors.forename: Takayuki
authors.initials: T
authors.lastname: Takahashi
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Iino
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamamoto
authors.forename: Kenji
authors.initials: K
authors.lastname: Takahashi
mesh_terms: D001483:Base Sequence; D002845:Chromatography; D017364:Collagenases / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D012689:Sequence Homology, Nucleic Acid; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D017364:Collagenases