[关键词]
[摘要]
以苦杏仁醇溶蛋白(BAKG)为原料,以α-葡萄糖苷酶活性抑制率及水解度为指标,筛选出酶解苦杏仁醇溶蛋白制备α-葡萄糖苷酶抑制酶解物(AGIH)的最佳蛋白酶,以α-葡萄糖苷酶活性抑制率为指标,采用单因素和响应面设计实验选定制备AGIH所需的底物浓度、加酶量、pH值、最佳温度、酶解时间。并对该最佳酶解条件下制备活性酶解物的物理(pH、温度)稳定性和胃肠道稳定性进行研究。结果表明,木瓜蛋白酶为最佳蛋白酶,酶解最优工艺条件为底物浓度4.0% (m/V)、加酶量6000 U/g、pH 7.0、温度55 ℃、酶解时间6 h。根据上述条件下制备的AGIH抑制率为18.10%,IC50为17.66 mg/mL。制得的AGIH在高温、低pH、高pH条件下,或者经胃肠道模拟消化后,都具有较好的稳定性。研究结果为苦杏仁醇溶蛋白功能活性酶解物的开发利用提供了新的思路。
[Key word]
[Abstract]
The optimal protease for preparing bitter almond kernel gliadin-based α-glycosidase inhibitory hydrolysates (AGIH) was determined according to the inhibition rate and degree of hydrolysis. Based on the α-glycosidase inhibition rate, optimal process parameters for AGIH preparation, including the substrate concentration required, amounts of enzymes added, pH, temperature, and duration, were examined in single-factor and response surface experiments. The physical stability (against pH and temperature) and gastrointestinal stability of the resulting AGIH were characterized. Papain was found to be the optimal protease; the optimal conditions were as follows: substrate concentration of 4.0% (m/V), enzyme addition of 6000 U/g, initial pH of 7.0, hydrolysis temperature of 55 ℃, and reaction time of 6 h. AGIH prepared under these conditions showed an α-glucosidase inhibition rate of 18.10% and IC50 value of 17.66 mg/mL, and was stable under relatively high temperature, extreme pH, and gastrointestinal digestion conditions. This study provides a foundation for developing bitter almond kernel gliadin-based bioactive hydrolysates.
[中图分类号]
[基金项目]
国家重点研发计划项目(2019YFD1002300)