title: Enzymatic properties of the recombinant serine-type carboxypeptidase OcpC, which is unique to Aspergillus oryzae.
abstract: Gene AO090103000153 is unique to Aspergillus oryzae RIB40 and A. flavus NRRL3357, and is speculated to encode a serine-type carboxypeptidase. In this study, we purified and characterized a heterologously expressed gene product of AO090103000153. 5'-Rapid amplification of cDNA ends indicated that the translation start site of the gene is located 1,586 bp downstream of the translation start site predicted by the genome sequencing project. The gene, starting from the revised translation start codon, termed ocpC, was transcribed constantly in A. oryzae RIB40. Purified recombinant OcpC exhibited the enzymatic properties of a serine-type carboxypeptidase. This protease was stable at temperatures below 45°C and a low pH, and had broad substrate specificity for N-acylpeptides, but it exhibited significantly lower specific activity and a lower k(cat) value for substrates than previously reported serine-type carboxypeptidases from A. oryzae.
authors.affiliation: Department of Applied Molecular Biology and Biochemistry, Tokyo University of Agriculture and Technology, Japan.
authors.forename: Hiroto
authors.initials: H
authors.lastname: Morita
authors.forename: Haruka
authors.initials: H
authors.lastname: Abo
authors.forename: Ayako
authors.initials: A
authors.lastname: Okamoto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Maeda
authors.forename: Youhei
authors.initials: Y
authors.lastname: Yamagata
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Kusumoto
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Amano
authors.forename: Hiroki
authors.initials: H
authors.lastname: Ishida
authors.forename: Michio
authors.initials: M
authors.lastname: Takeuchi
mesh_terms: D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000235:genetics; D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics; D002268:Carboxypeptidases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013045:Species Specificity; D014158:Transcription, Genetic
chemical_list: D011994:Recombinant Proteins; D002268:Carboxypeptidases; C022633:serine carboxypeptidase