具有ACE抑制活性的霞水母酶解肽制备条件优化

Study on Preparation of ACE Inhibitory Enzymatic Peptides of Cyanea

目的:采用胰蛋白酶制备霞水母ACE抑制肽,并以响应面法优化酶解肽制备工艺,通过超滤分离获得具有ACE抑制作用的酶解肽活性部位。方法:单因素实验以ACE抑制率为指标,考察温度、酶解时间、pH、加酶量,进一步通过响应面法优化酶解工艺,以不同截留分子量的纤维膜进行分离。结果:胰蛋白酶水解霞水母制备ACE抑制肽的最佳工艺条件为:酶解温度:49.6℃,加酶量为0.69%,pH为7.8,酶解时间2.0h,酶解产物的平均ACE抑制率为56.79%,与预测值57.20%的相对误差为0.41%,经过超滤分离获得分子量<1k Da的酶解产物,ACE抑制活性最高,其IC50为66.58μg/ml。结论:确定霞水母酶解肽的制备工艺,其中分子量<1kDa的部分为主要效应物质基础。

Objective： Trypsin was used to prepare ACE inhibitory peptides from Cyanea. Response surface method was used to optimize the ACE inhibitory peptides. The ultratihration membrane was subjected to obtain ACE inhibitory segments of Cyanea enzymatic hydrolysate. Methods： According to the ACE inhibitiory rate, temperature, reaction time, pH and enzyme dosage were chosen to optimize the hydrolysis conditions, then response surface design was furtherly used to optimize the enzymatic technology. Results： The best conditions were as follows ： hydrolysis temperature 49.6℃, enzyme dosage 0.69 %, hydrolysis pH 7.8, hydrolysis time 2 hours. The ACE inhibitory rate of hydrolysis peptides at best conditions was 56.79 %, compared with the theoretical value of 57.20 % , the relative error was 0.41%. The mass segment of Cyanea enzymatic hydrolysate obtained by ultratihration that molecular weight less than 1 kDa had the highest ACE inhibitory activity, which IC50 value is 66. 58μg/ml. Conclusion： The preparation of ACE inhibitory enzymatic peptides of Cyanea was determined and its material basis was the mass segment that molecular weight less than 1 kDa.