<研究論文>キノコプロテアーゼの精製と性質6. オオヒラタケ

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Two proteolytic enzymes, P. c. proteases 1 and 2, were purified from the fruit body of oohiratake mushroom, Pleurotus cystidiosus, by ammonium sulfate precipitation, gel-filtration, hydrophobic chromatography and ion-exchange chromatography. P. c. protease 1 was assayed using Boc-Ala-Ala-Pro-Phe-pNA as a substrate, and P. c. protease using Boc-Ala-Ala-Pro-Lys-pNA. The enzymes had the same optimal pH range of 8-9. 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 P. c. protease 1 preferencially hydrolyzed the peptide bonds of carboxyl-terminal side of phenylalanine and histidine, and preferred proline residue to leucine at P2 position of the substrate. On the other hand, P. c. protease 2 hydrolyzed the peptide bonds of many different amino acids including arginine, lysine, histidine and phenylalanine. Both enzymes were almost completely inactivated by diisopropyl phosphofluoridate, indicating the serine protease nature of the enzymes. While P. c. protease 2 was completely inhibited by chymostatin, leupeptin and antipain, P. c. protease 1 was not affected by these microbial protease inhibitors.
福岡女子大学の論文
2003-03-20