Purification and characterization of a collagenolytic serine proteinase from the skeletal muscle of red sea bream (Pagrus major)

Abstract: A collagenolytic serine proteinase (CSP) was purified from red sea bream (Pagrus major) skeletal muscle to homogeneity by ammonium sulfate fractionation and chromatographies including DEAE-Sephacel, Phenyl Sepharose and Hydroxyapatite. The molecular mass of CSP was approximately 85kDa as estimated by SDS–PAGE and gel filtration. Optimum temperature and pH of CSP were 40°C and 8.0, respectively. CSP was specifically inhibited by serine proteinase inhibitors, while inhibitors to other type proteinases did not show much inhibitory effects. The K m and k cat values of CSP for Boc-Leu-Lys-Arg-MCA were 3.58µM and 0.13s−1 at 37°C, respectively. Furthermore, CSP hydrolyzed gelatin and native type I collagen effectively though its degradation on myosin heavy chain (MHC) was not significant, suggesting its involvement in the texture tenderization of fish muscle during the post-mortem stage. [Copyright &y& Elsevier]