南瓜籽粕酶法制备血管紧张素转化酶抑制肽工艺优化

Process optimization for preparation of ACE inhibition peptide from pumpkin seed meal by using enzymatic hydrolysis method

用中性蛋白酶水解南瓜籽粕,制备血管紧张素转换酶(angiotensin—converting enzyme,ACE)抑制肽,是南瓜籽粕蛋白深度开发的途径之一。为了探寻南瓜籽粕酶法制备ACE抑制肽的最佳工艺,该文以ACE抑制率为响应值,用响应面分析法研究酶浓度、底物质量浓度和水解时间等因素对酶解产物的ACE抑制活性的影响,优化制备工艺。结果表明,各因素对制备ACE抑制肽的活性具有极显著影响(P<0.001)。获得中性蛋白酶水解南瓜籽粕制备ACE抑制肽的最佳工艺条件为:酶体积分数为4.8%、底物质量浓度为4.0g/100 m L、水解时间为320 min。在此条件下,南瓜籽粕蛋白酶解产物对ACE的理论抑制率最高可达80.0%,验证值为80.56%±0.23%,预测模型可靠性高,可应用于南瓜籽粕ACE抑制肽的酶法制备。通过优化,提高了南瓜籽粕ACE抑制肽的活性。

Angiotensin-converting enzyme (ACE) inhibitory peptides made from hydrolysis of pumpkin seed meal with neutral protease is one of the important ways to utilization pumpkin seed meal. In order to find the optimum craft of ACE inhibitory peptides made from pumpkin seed meal by protease, taking ACE inhibitory rate as the response value, protease content,the effects of substrate content,and hydrolysis time on ACE inhibitory activity of the hydrolysates were analyzed with response surface methodology for optimization of the preparation technology.The results showed that the three factors affecting activity of ACE inhibitory peptides significantly (P<0.001). The optimized parameters for preparation of angiotensin-converting enzyme inhibitory peptides were temperature at 45℃ and pH value of 7.0, the protease content 4.8%, substrate content of 4.0 g/100mL, and hydrolysis time of 320 min. With the optimal parameters, the ACE inhibiting percentage theoretically could be up to 80%. The validation test showed that the actual ACE inhibiting percentage was 80.56%±0.23%, which was closer to 80%. It suggested that the established prediction model is reliable. The results here proposed the viable and reliable optimization parameters for preparation of ACE inhibitory peptides.