Comparative Biochemical Properties of Proteinases from the Hepatopancreas of Shrimp. -II. Purification of Trypsin from the Hepatopancreas of Penaeus orientalis-

  • Oh Eun-Sil (Department of Food Science and Nutrition, Yosu National University) ;
  • Kim Doo-Sang (Department of Food Science and Nutrition, Yosu National University) ;
  • Jung Kyoo-Jin (Fisheries Research Institute of Chunnam) ;
  • Pyeun Jae-Hyeung (Department of Food and Life Science, Pukyong National University) ;
  • Heu Min-Soo (Department of Food Science, Gyeongsang National University) ;
  • Kim Hyeung-Rak (Department of Food Science and Nutrition, Yosu National University)
  • Published : 1998.12.01

Abstract

Trypsin-like enzyme was purified from shrimp hepatopancreas through Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, and Superdex 75 gel chromatography. Purity of trypsin-like enzyme was increased 69-fold with $44\%$ yield. The enzyme consisted of a single polypeptide chain with a molecular weight (M.W.) of 32 kDa judged by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was completely inactivated by serine enzyme inhibitors such as soybean trypsin inhibitor (SBTI), tosyl-L­lysine chloromethyl ketone (TLCK), and leupeptin. However, the enzyme was not affected by tosyl-L-phenylalanine chloromethyl ketone (TPCK) which is a chymotrypsin specific inhibitor. The enzyme had no activity against benzoyl-tyrosine ethyl ester (BTEE) which is a chymotrypsin specific substrate. The enzyme showed high activity on the carboxyl terminal of Phe, Tyr. Glu, Arg, and Asp. However. no activity was detected against the carboxyl terminal of Pro, Trp, Cys, Gly, Val, and Ala.

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