title: 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 85 kDa as estimated by SDS-PAGE and gel filtration. Optimum temperature and pH of CSP were 40 degrees 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 3.58 microM and 0.13 s(-1) at 37 degrees 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.
authors: Fisheries College, The Key Laboratory of Science and Technology for Aquaculture and Food Safety, Jimei University, Xiamen 361021, China.
authors: Guo-Ping
authors: GP
authors: Wu
authors: Su-Hua
authors: SH
authors: Chen
authors: Guang-Ming
authors: GM
authors: Liu
authors: Asami
authors: A
authors: Yoshida
authors: Ling-Jing
authors: LJ
authors: Zhang
authors: Wen-Jin
authors: WJ
authors: Su
authors: Min-Jie
authors: MJ
authors: Cao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003094:Collagen / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000494:pharmacology; D018482:Muscle, Skeletal / Q000201:enzymology*; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D010473:Perciformes; D057057:Serine Proteases / Q000302:isolation & purification* / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D008670:Metals; D009842:Oligopeptides; D015842:Serine Proteinase Inhibitors; D003094:Collagen; D057057:Serine Proteases