Hydrolytic properties of crude protease from Bacillus subtilis subsp. subtilis M13

Abstract

A new strain of Bacillus subtilis subsp. subtilis M13 which isolated from meat is proved to be the collagenase producing bacteria based on high degradation of denatured form of collagen or gelatin. The molecular weight of the crude enzyme was approximately be 21 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Maximum collagenolytic activity based on gelatin as a substrate was attained at 50°C and pH 6.0 with citrate buffer. Furthermore, this enzyme was preferable to hydrolyze insoluble collagen, and followed by elastin at 37°C and pH 6.0. Maximum hydrolysis for collagen was showed by the highest release of amino acid at 998 mg/ml after incubation for 24 h, and elastin at 656 mg/ml after incubation for 12 h. However, the hydrolytic activity against myofibrillar protein extracted from meat was relatively lower than 3.3 mg/ml at 6 h. There was found either no or slightly hydrolytic effect after prolonged incubation. The results indicated the potential protease for using as meat tenderizing enzyme with high degradation of collagen and elastin with low hydrolysis of meat myofibrillar protein.