title: Purification and characterization of a novel dipeptidyl carboxypeptidase from a Streptomyces species.
abstract: An extracellular protease derived from the culture broth of a microorganism, a Streptomyces species, produced Boc-Pro-Pro and diproline from Boc-Pro-Pro-Pro-Pro. The enzyme was purified 726-fold, with a yield of 2.6%, by ammonium sulfate fractionation, ion-exchange chromatography, and gel filtration chromatography. The molecular weight of the enzyme was determined to be 65,000 by gel filtration and 70,000 by SDS-PAGE. The enzyme released a C-terminal dipeptide from peptide substrates having a C-terminal proline and a penultimate proline or alanine residue, but did not hydrolyze angiotensin I or bradykinin. When the enzyme hydrolyzed Leu-Pro-Pro-Pro-Pro-Pro, it produced Leu-Pro-Pro-Pro and Pro-Pro before producing Leu-Pro. The enzyme thus seems to be a kind of dipeptidyl carboxypeptidase, its substrate specificity being very different from that of the well known dipeptidyl carboxypeptidases [EC 3.4.15.1] such as the angiotensin-converting enzyme.
authors.affiliation: Research and Development Center, Showa Sangyo Co., Ltd., Chiba.
authors.forename: S
authors.initials: S
authors.lastname: Miyoshi
authors.forename: G
authors.initials: G
authors.lastname: Nomura
authors.forename: M
authors.initials: M
authors.lastname: Suzuki
authors.forename: F
authors.initials: F
authors.lastname: Fukui
authors.forename: H
authors.initials: H
authors.lastname: Tanaka
authors.forename: S
authors.initials: S
authors.lastname: Maruyama
mesh_terms: D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009842:Oligopeptides / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D009842:Oligopeptides; D010450:Endopeptidases; C080173:proline-specific dipeptidylcarboxypeptidase