Purification,Characterization and cDNA Cloning of a Myofibril-bound Serine Proteinase from the Skeletal Muscle of Crucian Carp(Carassius auratus)

作     者：Chuan Guo~a Min-Jie Cao~(a*) Guang-Ming Liu~a Xiong-Shui Lin~a Kenji Hara~b Wen-Jin Su~a (~a College of Biological Engineering,The Key Laboratory of Science and Technology for Aquaculture and Food Safety,Jimei University,Xiamen,China,361021 ~bFaculty of Fisheries,Nagasaki University,Nagasaki,Japan,852-8521) 

会议名称：《中国食品科学技术学会第五届年会暨第四届东西方食品业高层论坛》

会议日期：2007年

学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 

关 键 词：Crucian carp Myofibril Serine proteinase cDNA cloning 

摘      要：A myofibril-bound serine proteinase(MBSP)was highly purified from the skeletal muscle of crucian carp(carasius auratus)by acidic treatment of myofibril solution and chromatographies on Q-Sepharose and benzamidine-Sepharose *** revealed a main protein band of approximately 28 kDa on SDS-PAGE and was particularly inhibited by serine proteinase *** specificity analysis revealed that the enzyme specifically cleaved at the carboxyl side of arginine and lysine residues,suggesting the characteristic of a trypsin-type serine *** gene was cloned based on the N-terminal sequence and the conserved active site peptide of serine proteinases together with 5’-RACE and 3’-*** coding region gave an amino-acid sequence of 242 residues including the initiation methionine and a signal peptide of 20 *** acid residues of His,Aspand Serconsisting the catalytic triad of serine proteinases were conserved in the *** carp MBSP shared relatively high identities with other serine proteinases,especially in well-conserved regions.