草鱼鱼鳞胶原蛋白酸酶分步提取工艺研究

Combinated use of enzyme and acid for collagen extraction from fish scales

为实现鱼鳞胶原蛋白的高效提取,在优化脱钙、混酸法和酶法提取工艺基础上,对鱼鳞胶原蛋白的酸酶进行分步提取。研究发现,鱼鳞最佳脱钙工艺为料液比8:100(g/mL),盐酸浓度1.0 mol/L,反应时间1.0 h,反应温度20℃;混合酸法提取鱼鳞胶原蛋白的最佳条件为醋酸料液比1:12(g/mL),柠檬酸料液比1:10(g/mL),乳酸料液比1:12(g/mL),即混合酸料液比为1:34(g/mL),其中,0.8 mol/L柠檬酸、1 mol/L乳酸、0.8 mol/L醋酸的体积比为6:5:6,提取时间2 d,胶原蛋白的提取率为48.14%;最佳酶法提取条件为胃蛋白酶用量450 U/g,提取温度30℃,提取时间72 h,该条件下提取率为45.26%。酸酶耦合法优于单一方法或同种方法两次提取的效果,可实现酸溶性和酶溶性胶原蛋白的连续提取,先酸后酶法胶原蛋白的提取率达84.61%,SDS-PAGE凝胶电泳发现其为Ⅰ型胶原蛋。

In order to realize the efficient extraction of collagen from fish scales, acid-enzyme coupling methods was used for collagen ex- traction based on the optimized decalcification, mixed-acid and enzy matic extraction. The optimum decalcification of fish scales was as follows. The solid to liquid ratio was 8 ： 100, soluted in 1.0 mol/L hydrochloric acid, and then reaction at 20℃ for 1.0 h. The beat con- ditions for collagen extraction using mixed acid were 1 ： 12 of acetic acid, 1 ： 10 of citric acid, and 1 ： 12 of lactic acid for 3 d extraction and the extraction yield of it was 48.14%0. The best enzymatic extrac- tion conditions were shown to be 450 U/g of pepsin at the extraction temperature of 30℃ for 72 h and the yield was 45.26%. Acid- enzyme coupling method showed better effeet than acid or enzymatic extraction only, even though repeating them twice respectively was not as effective as the coupled one, in spite of relatively high cost and time-consuming （P〈0.05）. We could obtain 84.61% of the collagen using mixed acid followed by enzymatic extraction , and the extracted collagen was identified to be type I one using SDS-PAGE.