title: Gene cloning and biochemical characterization of eryngase, a serine aminopeptidase of Pleurotus eryngii belonging to the family S9 peptidases.
abstract: Pleurotus eryngii serine aminopeptidase that has peptide bond formation activity, redesignated as eryngase, was cloned and expressed. Eryngase has a family S9 peptidase unit in the C-terminal region having a catalytic triad of Ser, Asp, and His. In the phylogenetic relations among the subfamilies of family S9 peptidase (S9A, prolyl oligopeptidase; S9B, dipeptidyl peptidase; S9C, acylaminoacyl peptidase; S9D, glutamyl endopeptidase), eryngase existed alone in the neighbor of S9C subfamily. Mutation of the active site Ser524 of the eryngase with Ala eliminated its catalytic activity. In contrast, S524C mutant maintained low catalytic activity. Investigation of aminolysis activity using l-Phe-NH2 as a substrate showed that S524C mutant exhibited no hydrolysis reaction but synthesized a small amount of l-Phe-l-Phe-NH2 by the catalysis of aminolysis. In contrast, wild-type eryngase hydrolyzed the product of aminolysis l-Phe-l-Phe-NH2. Results show that the S524C mutant preferentially catalyzed aminolysis when on an l-Phe-NH2 substrate.
authors: a Faculty of Agriculture, Department of Agricultural, Biological, and Environmental Sciences , Tottori University , Tottori , Japan.
authors: Jiro
authors: J
authors: Arima
authors: Shota
authors: S
authors: Tokai
authors: Masanori
authors: M
authors: Chiba
authors: Tsuyoshi
authors: T
authors: Ichiyanagi
authors: Yukinori
authors: Y
authors: Yabuta
authors: Nobuhiro
authors: N
authors: Mori
authors: Tadanori
authors: T
authors: Aimi
mesh_terms: D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D020134:Catalytic Domain / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D008969:Molecular Sequence Data; D009154:Mutation; D010447:Peptide Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020076:Pleurotus / Q000145:classification / Q000201:enzymology* / Q000235:genetics*; D016415:Sequence Alignment; D012694:Serine / Q000737:chemistry / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D012694:Serine; D010447:Peptide Hydrolases; D000626:Aminopeptidases
keywords: Pleurotus eryngii; family S9 peptidase; peptide bond formation; serine aminopeptidase