title: Molecular cloning of ocpO encoding carboxypeptidase O of Aspergillus oryzae IAM2640.
abstract: Carboxypeptidase O from Aspergillus oryzae IAM2640 is a serine-type carboxypeptidase. In this study, we cloned and sequenced cDNA and genomic DNA carrying ocpO encoding carboxypeptidase O. The results showed that the length of ocpO was 1,816 bp, and the open reading frame encoded a putative preproenzyme composed of 472 amino acid residues of the mature carboxypeptidase O and an additional N-terminal sequence of 50 amino acid residues. A BLASTN search revealed that a gene, AO090020000351, in A. oryzae RIB40, which is strain used in genome-wide sequencing, is a homolog of ocpO. The difference between AO090020000351 and ocpO was only one nucleotide. The difference caused substitution of Ala for Pro at the 277th position of the enzyme; therefore the protein encoded by AO090020000351 was overproduced and purified. The purified protein showed enzymatic properties similar to carboxypeptidase O, indicating that carboxypeptidase O and protease encoded by AO090020000351 are same enzyme.
authors.affiliation: Department of Applied Molecular Biology and Biochemistry, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, Japan.
authors.forename: Hiroto
authors.initials: H
authors.lastname: Morita
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Kuriyama
authors.forename: Noritaka
authors.initials: N
authors.lastname: Akiyama
authors.forename: Ayako
authors.initials: A
authors.lastname: Okamoto
authors.forename: Youhei
authors.initials: Y
authors.lastname: Yamagata
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Kusumoto
authors.forename: Yoshinao
authors.initials: Y
authors.lastname: Koide
authors.forename: Hiroki
authors.initials: H
authors.lastname: Ishida
authors.forename: Michio
authors.initials: M
authors.lastname: Takeuchi
mesh_terms: D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D002268:Carboxypeptidases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D030541:Databases, Genetic; D004795:Enzyme Stability; D016681:Genome, Fungal / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000494:pharmacology; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity
chemical_list: D018076:DNA, Complementary; D004271:DNA, Fungal; D011480:Protease Inhibitors; D002268:Carboxypeptidases