Waite J H, Tanzer M L, Merkel J R
J Biol Chem. 1980 Apr 25;255(8):3596-9.
Proteolysis of Nereis cuticle collagen by two bacterial collagenases was investigated using viscosimetry, enzyme kinetics, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and ion exchange chromatography of collagenolytic peptides. Collagenase of the marine Vibrio B-30 completely degrades native cuticle collagen at 7 degress C with a turnover number 50 times greater than that of the clostridial collagenase. Although turnover numbers for the two enzymes are comparable when using denatured cuticle collagen as substrate, the vibrial collagenase appears to cleave twice as many peptide bonds per mg of cuticle collagen as does the clostridial enzyme. Sodium dodecyl sulfate gel electrophoresis of collagenase-digested native cuticle collagen reflects the resistance of the collagen to clostridial collagenase; however, the vibrial enzyme completely degrades the cuticle collagen with the formation of one transient intermediate (Mr 400,000). Peptide analysis of fully digested denatured cuticle collagen reveals that the two enzymes have a number of qualitative and quantitative similarities. Despite these, however, only the vibrial collagenase seems capable of extensively degrading native cuticle collagen.
利用粘度测定法、酶动力学、十二烷基硫酸钠聚丙烯酰胺凝胶电泳以及胶原水解肽的离子交换色谱法，研究了两种细菌胶原酶对沙蚕表皮胶原蛋白的蛋白水解作用。海洋弧菌B - 30的胶原酶在7摄氏度时能完全降解天然表皮胶原蛋白，其转换数比梭菌胶原酶的转换数大50倍。虽然以变性表皮胶原蛋白为底物时，两种酶的转换数相当，但弧菌胶原酶每毫克表皮胶原蛋白所切割的肽键数量似乎是梭菌酶的两倍。胶原酶消化的天然表皮胶原蛋白的十二烷基硫酸钠凝胶电泳反映了胶原蛋白对梭菌胶原酶的抗性；然而，弧菌酶能完全降解表皮胶原蛋白，并形成一种瞬时中间体（分子量400,000）。对完全消化的变性表皮胶原蛋白的肽分析表明，这两种酶在定性和定量方面有许多相似之处。然而，尽管如此，只有弧菌胶原酶似乎能够广泛降解天然表皮胶原蛋白。
