Isolation and characterization of trypsin from fish viscera of Oil Sardine (Sardinella longiceps)

Abstract

The development of the fisheries industry has led to the increased interest in the full utilization of non-edible seafood fish viscera as a large source of unique digestive enzymes especially the proteases. We report the purification and biochemical characterization of a trypsin from the viscera of Oil Sardine (Sardinella longiceps) using anion-exchange and soybean trypsin inhibitor (SBTI) affinity chromatography. Final enzyme preparation was homogeneous in SDS-PAGE and the molecular weight of the purified enzymes was estimated to be 24,000 Da. The enzyme activity was optimum at 60 °C for hydrolysis of benzoyl-dl-arginine-p-nitroanilide (BAPNA). The purified enzyme maximal activity was observed at pH 8.0. Isolated trypsin was strongly inhibited by SBTI and N-p-tosyl-1-lysine chloromethyl ketone (TLCK) which are the specific inhibitors of trypsin and was markedly inhibited by the serine-protease inhibitor PMSF. Results signify that Oil sardine viscera are useful source for production of enzymes that could be used as a biotechnological tool in various industries.