Comparative Biochemical Properties of Proteinases from the Hepatopancreas of Shrimp. -I. Purification of Protease from the Hepatopancreas of Penaeus japonicus-

  • Choi Sung-Mi (Department of Food Science and Nutrition, Yosu National University) ;
  • Oh Eun-Sil (Department of Food Science and Nutrition, Yosu National University) ;
  • Kim Doo-Sang (Department of Food Science and Nutrition, Yosu National University) ;
  • Pyeun Jae-Hyeung (Department of Food and Life Science, Pukyong National University) ;
  • Cho Deuk-Moon (Department of Food and Nutrition, Dong-Pusan University) ;
  • Ahn Chang-Bum (Department of Food Science and Nutrition, Yosu National University) ;
  • Kim Hyeung-Rak (Department of Food Science and Nutrition, Yosu National University)
  • Published : 1998.12.01

Abstract

A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

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