具有α-葡萄糖苷酶抑制活性的苦杏仁醇溶蛋白酶解物制备及稳定性表征

Preparation and Stability Characterization of Bitter Almond Kernel Gliadin Hydrolysate with α-Glucosidase Inhibitive Activity

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% (w/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。

Abstract:

Bitter almond kernel gliadin (BAKG)-based α-glycosidase inhibitive hydrolysates (AGIH) was prepared by protease catalyzed hydrolysis of BAKG using the inhibition rate and the degree of hydrolysis as indicators. The optimal protease was determined, the optimal process parameters was established via single-factor and response surface experiments, and the pH, thermal, and gastrointestinal digestive stability of the resultant AGIH was characterized. Papain was determined as the optimal protease, with the pertinent optimum conditions established as: solid-liquid ratio 4.0% (w/v), enzyme addition 6000 U/g, initial pH 7.0, hydrolysis temperature 55 ℃, and reaction time 6 h. The AGIH prepared under optimum conditions showed an α-glucosidase inhibition rate of 18.10% and an IC50 value of 17.66 mg/mL, and exhibited good stability under relatively high temperature, extreme pH and gastrointestinal digestion conditions. This study provides a novel direction for developing BAKG based bioactive hydrolysates.