Author(s): Ajay S Khandagale, Likhitha Mundodi and Balladka Kunhanna Sarojini

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.

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