title: Grifolisin, a member of the sedolisin family produced by the fungus Grifola frondosa.
abstract: The pepstatin-insensitive carboxyl proteinase grifolisin was purified from fruiting bodies of the fungus Grifola frondosa, a maitake mushroom. The enzyme had an optimum pH of 3.0 for the digestion of hemoglobin and 2.8 for milk casein digestion. Its molecular mass was determined to be 43kDa by SDS-PAGE and 40kDa by gel chromatography on Superose 12, and its isoelectric point was found to be 4.6 by isoelectric focusing. The enzyme hydrolyzed four major bonds in the oxidized insulin B-chain: Phe1-Val2, Ala14-Leu15, Gly20-Glu21 and Phe24-Phe25 at pH 3.0. The first 15 amino acid residues in the N-terminal region were AVPSSCASTITPACL, and the coding region of the grifolisin gene (gfrF) has a 1960-base pair cDNA. The predicted mature grifolisin protein consisted of 365 residues and was 26% identical to that of sedolisin from Pseudomonas sp. 101 and 34% identical to that of aorsin from Aspergillus oryzae. Grifolisin is a member of the sedolisin S53 family and is not inhibited by pepstatin.
authors.affiliation: Laboratory of Molecular Enzymology, Department of Bioengineering, Faculty of Engineering, Soka University, 236 Tangi-cho, Hachioji-Shi, Tokyo 192-8577, Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Suzuki
authors.forename: Kohzoh
authors.initials: K
authors.lastname: Nishibori
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Oodaira
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Kitamura
authors.forename: Kenji
authors.initials: K
authors.lastname: Michigami
authors.forename: Keiko
authors.initials: K
authors.lastname: Nagata
authors.forename: Yota
authors.initials: Y
authors.lastname: Tatara
authors.forename: Byung Rho
authors.initials: BR
authors.lastname: Lee
authors.forename: Eiji
authors.initials: E
authors.lastname: Ichishima
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002268:Carboxypeptidases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D048691:Grifola / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D002268:Carboxypeptidases; C504935:grifolisin, Grifola frondosa