Purification and Characterization of Protease from the Hepatopancreas of Shrimp, 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) ;
  • Choi Sung-Mi (Department of Food Science and Nutrition, Yosu National University) ;
  • Kim Jeong-Han (Department of Refrigeration Engineering, Yosu National University) ;
  • Pyeun Jae-Hyeung (Faculty of Food and Biotechnology, Pukyong National University) ;
  • Cho Deuk-Moon (Department of Food and Nutrition, Dong-Pusan College) ;
  • Kim Hyeung-Rak (Department of Food Science and Nutrition, Yosu National University)
  • Published : 1999.12.01

Abstract

A protease without tryptic and chymotryptic activities was purified from the hepatopancreas of shrimp, Penaeus orientalis, using Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, Mono-Q, and gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 27kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS­PAGE). The amino acid composition of the protease was different from that of protease from P. japonicus or trypsin from P. orientalis. The protease was completely inhibited by benzamidine, $N\alpha-p-tosyl-L-lysine$ chloromethyl ketone (TLCK), and phenylmethylsulfonyl fluoride (PMSF) and was not affected by leupeptin, pepstatin, N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), iodoacetate, and ethylenediamine tetra acetate (EDTA). The enzyme did not have any activity against Na-benzoyl-DL-arginine p-nitroanilide (BAPNA) or N-benzoyl-L-tyrosine ethyl ester (BTEE) which are specific substrates of trypsin and chymotrypsin, respectively. However, the protease showed hydrolytic activity for a carboxyl terminal of Tyr, Trp, Phe, Glu, and Cys.

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