双酶法制备哈蟆油蛋白多肽的研究

Study on Oviductus Ranae Polypeptide by Double Enzymes

采用碱性蛋白酶和胃蛋白酶酶解哈蟆油制备蛋白多肽。应用单因素和正交试验考察碱性蛋白酶用量、酶解时间,胃蛋白酶用量、酶解时间,依据酶解后哈蟆油蛋白多肽分子量小于5 kDa所占比例,优选出制备哈蟆油蛋白多肽的最佳工艺。确定最佳制备工艺为碱性蛋白酶与哈蟆油比为1︰500(g/g),酶解时间为9 h;胃蛋白酶与哈蟆油比为1︰10 000(g/g),酶解时间为3 h。酶解得到的哈蟆油蛋白多肽相对分子量小于5 kDa的约占97.10%。

Oviductus ranae was hydrolyzed by alkaline protease and pepsin. The effects of alkaline pmtr^ase on enzymatic degree,enzymatic time, pepsin on enzymatic degree, enzymatic time were optimized by orthogonal design and single factor tests accordingto the amount of oviductus ranae polypeptide by enzymes proportion less than 5 kDa to determine enzymatic conditions. Theoptimum enzymatic condition of oviductus ranae protein isolate hydrolyzed with alkaline protease substrate ratio l 500 （g/g）, enzymatic time 9 h; pepsin substrate ratio of 1 ： 10 000 （g/g）, enzymatic time 3 h. The amount of the oviductus ranaepolypcptide prepared by hydrolyze less than the proportion of 5 kDa was 97.10%.