摘要:
通过响应面法优化裙带菜α-葡萄糖苷酶抑制活性肽的制备工艺,以期得到一种调控餐后血糖的新型有效成分。选择五种蛋白酶酶解裙带菜蛋白筛选最佳水解酶,研究底物浓度、加酶量、pH、酶解温度、酶解时间对产物抑制率和水解度的影响,并根据单因素实验结果运用Box-Behnken设计原理进行三因素三水平的响应面优化试验,测定酶解液的分子量并绘制酶抑制动力学曲线。结果表明,最佳酶解条件为碱性蛋白酶,底物浓度7.11%,pH10.14,温度47 ℃,加酶量10000 U/g,反应时间1 h,在此条件下,酶解液的抑制率为51.17%,与预测值接近;裙带菜酶解液多为小肽,半抑制浓度为46.079 mg/mL,抑制类型为典型的可逆混合型抑制。本研究获得了裙带菜α-葡萄糖苷酶抑制活性肽的最佳制备工艺和理化性质,为开发新型降血糖活性肽提供理论基础和实验依据。
Abstract:
The preparation process of α-glucosidase inhibitory active peptides from sea mustard was optimized by response surface method in order to obtain a new effective ingredient for regulating postprandial blood glucose. Five proteases were selected to hydrolyze sea mustard protein. The effects of substrate concentration,enzyme-substrate ratio,pH,temperature,and hydrolysis time on inhibition rate and degree of hydrolysis were studied. Based on the results of single-factor experiments,Box-Behnken design principle was used to perform a three-factor and three-level response surface optimization test. Then the molecular weight of the hydrolysate was measured and the kinetic curve of enzyme inhibition was drawn. The optimal hydrolysis conditions were:Alcalase,substrate concentration 7.11%,pH10.14,temperature 47 ℃,enzyme concentration 10000 U/g,and reaction time 1 h.Under these conditions,the inhibition rate was 51.17%. The half-inhibitory concentration of the sea mustard hydrolysate was 46.079 mg/mL,and the inhibition type was typical reversible mixed-type inhibition. In this study,the optimal preparation technology and physicochemical properties of α-glucosidase inhibitory active peptides from sea mustard were obtained,which provided theoretical and experimental basis for the development of novel hypoglycemic active peptides.
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