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Abstract: Trypsins, TR-S and TR-P, from the viscera of true sardine (Sardinops melanostictus) and from the pyloric ceca of arabesque greenling (Pleuroprammus azonus), respectively, were purified by gel filtration and anion-exchange chromatography. Final enzyme preparations were nearly homogeneous in sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and the molecular weights of both enzymes were estimated to be 24,000 Da by SDS–PAGE. The N-terminal amino acid sequences of the TR-S, IVGGYECKAYSQPWQVSLNS, and TR-P, IVGGYECTPHTQAHQVSLNS, were found. The TR-S and TR-P had maximal activities at around pH 8.0 for hydrolysis of N α-p-tosyl-l-arginine methyl ester. Optimum temperature of the TR-S and TR-P were 60 and 50°C, respectively. The TR-S and TR-P were unstable at above 50 and 30°C, respectively, and below pH 5.0. Both TR-S and TR-P were stabilized by calcium ion. [Copyright &y& Elsevier]

Abstract: Carboxypeptidase B was purified from the pyloric ceca of the starfish, Asterina pectinifera. The final enzyme preparation was nearly homogeneous in sodium dodecyl sulfate–polyacrylamide gel electrophoresis and its molecular weight was estimated as approximately 34,000. The value of the specificity constant (k cat/K m) for hydrolysis of benzoyl-glycyl-l-arginine by the purified enzyme was 1.72×105 M−1 s−1. The optimal pH and the optimal temperature of the enzyme were pH 7.5 and 55°C, respectively. The enzyme was unstable above 50°C and below pH 5.0. The enzyme was activated by Co2+, and inhibited by EDTA. The N-terminal amino acid sequence of the enzyme was determined as ATFDYNKYHSYQEIMDWVTN. [Copyright &y& Elsevier]