<研究論文>キノコプロテアーゼの精製と性質5. ウスヒラタケ

概要

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An endoprotease was partially purified from the homogenate of fruit body of usuhiratake mushroom, Pleurotusflavellatus, by ammonium sulfate precipitation, gel-filtration on Toyopearl HW55, hydrophobic chromatography on Butyl-Toyopearl and ionexchange chromatography on DEAE-Toyopearl using Boc-Ala-Ala-Pro-Phe-pNA as a substrate. Studies on the substrate specificity using a series of synthetic tetrapeptide p-nitroanilides having the sequence Boc-Ala-Ala-Pro-X-pNA and Boc-Ala-Ala-Leu-X-pNA (X = amino acids) revealed that the protease hydrolyzed the peptide bonds on carboxyl-terminal side of phenylalanine, histidine, and asparagines with proline residue at P_2 subsite. Peptide p-nitroanilides with leucine residue at P_2 position were not hydrolyzed by the enzyme. The activity was completely inactivated by diisopropyl phosphofluoridate (DFP) and chymostatin, and strongly inhibited by tosyl-L-phenylalanine chloromethylketone (TPCK). From the primary substrate specificity and inactivation behavior, the enzyme was presumed to ba a chymotrypsin-like serine protease.
福岡女子大学の論文
2003-03-20