title: Production, purification, and properties of serine carboxypeptidase from Paecilomyces carneus.
abstract: Seventeen strains of the genus Paecilomyces were examined for their ability to produce serine carboxypeptidase. Paecilomyces carneus IFO 7012 exhibited the highest potency for serine carboxypeptidase production. A maximum yield of serine carboxypeptidase was obtained by koji culture of the strain at 22 degrees C for 7 days. The serine carboxypeptidase was purified to homogeneity from an extract of the koji culture. The molecular weight of the enzyme was estimated to be 47,000 by HPLC. The isoelectric point of the enzyme was determined to be 4.0, and the optimum pH was 4.0 toward benzyloxycarbonyl-L-glutamyl-L-tyrosine (Z-Glu-Tyr) and benzyloxycarbonyl-L-phenylalanyl-L-alanine (Z-Phe-Ala), respectively. The enzyme was strongly inhibited by phenylmethylsulfonyl fluoride and p-chloromercurybenzoate. Relative hydrolysis rates of N-acylpeptides and kinetic studies indicated that the enzyme preferred substrates having bulky amino acids in the penultimate position from their carboxy-termini.
authors.affiliation: Institute of Bioscience and Biotechnology, Aomori University, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Umetsu
authors.forename: K
authors.initials: K
authors.lastname: Hishinuma
authors.forename: H
authors.initials: H
authors.lastname: Wake
authors.forename: E
authors.initials: E
authors.lastname: Ichishima
mesh_terms: D000595:Amino Acid Sequence; D002268:Carboxypeptidases / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D002729:Chloromercuribenzoates / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009842:Oligopeptides / Q000737:chemistry; D010143:Paecilomyces / Q000201:enzymology*; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D013379:Substrate Specificity
chemical_list: D002729:Chloromercuribenzoates; D004791:Enzyme Inhibitors; D009842:Oligopeptides; D010664:Phenylmethylsulfonyl Fluoride; D002268:Carboxypeptidases; C022633:serine carboxypeptidase