Alcalase酶解蛋清粉制备抗凝血肽的工艺优化

Optimization of preparation for anticoagulant peptide from egg white powder by Alcalase

采用Alcalase碱性蛋白酶酶解蛋清粉制备食源性抗凝血肽。结果表明:底物浓度过大会抑制水解度的增长,底物质量分数为1%时蛋白质完全水解后的产物抗凝血活性最佳;高温和低温均对水解度有影响,低温酶解产物的抗凝血活性较好;pH值对水解度的影响与Al-calase的最适pH值有关;pH值为7时,抗凝血活性最好;酶/底物浓度增大,但底物不变的情况下,对水解度的提高不明显,酶/底物浓度过大,导致抗凝血活性降低。考虑交互作用经试验优化设计确定的酶解最佳工艺参数为:底物质量分数为1%,酶解温度为50℃,酶/底物质量分数为5%,酶解pH为8。在该条件下水解2h,凝血抑制率达到68.92%。

Anticoagulant peptide was prepared by hydrolyzing egg white powder with Alcalase.Results show that the degree of hydrolysis（DH） was inhibited by over-high substrate concentration.Anticoagulant activity was optimum when the substrate concentration was 1%.Temperature strongly affects DH that at low temperature the hydrolyzed possesses high anticoagulant activity.The effect of pH on DH was related to the optimum pH value of Alcalase.When pH value was 7 the anticoagulant activity was the best.DH increased slowly and anticoagulant activity decreased with excessive increase in enzyme dose.The optimal technical parameters were： substrate concentration of 1%,temperature of 50 °C,enzyme concentration of 5% and pH value of 8.After 2 h hydrolization the anticoagulant activity reached 68.92%.