双酶分步水解制备棉籽多肽工艺条件优化

Optimization of stepwise dual-enzymatic preparation of cottonseed polypeptides

为高效制备棉籽多肽,采用Alcalase酶和Flavourzyme酶对棉籽蛋白进行双酶分步水解。在前期单因素试验的基础上,通过Plackett-Burrman设计对影响其多肽得率的相关因素进行评估并筛选出具有显著效应的3个因素,然后通过三因素三水平的Box-Behnken响应面分析法确定双酶分步水解棉籽蛋白的最优操作条件为:底物浓度(w/v)8.36%,先用Alcalase酶在温度59.9℃、pH值8.0、酶用量25 000 U/g的条件下水解150 min,再用Flavourzyme酶在温度50℃、pH值6.0、酶用量27 430 U/g的条件下水解120 min,多肽得率可达到65.07%。

Cottonseed protein was hydrolyzed by Alacalase anti FlawJurzyme to produce active peptides. Through Plackett-Burrman design, some process factnrs under hydrolysis conditions were evaluated anti three factors had prominent effect on yiehl of cottonseed peplides were picked out, which were substrate concentration（w/v）, temperature of alacalase hydrolysis and amount of Flavorzyme respectively. By Box-Behnken design and response surface analysis, the optimal hydrolysis conditions： substrate concentration 8.36% （w/v）, firstly, Alacalase hydrolyzed for 150 min ureter the conditinns of 59.9 ℃, pH 8.0 and enzyme/suhstrate 25 000 U/g, then Flavorzyme hydrolyzed for 120 min under the conditions of 50 ℃, pH 6.0 anti enzyme/stlhslrale 27 430 U/g, under these condition, cottonseed peplides with the yield of 65.07% were prepared.