绿豆蛋白降血脂水解物的制备及纯化工艺

以碱提酸沉法制备的绿豆蛋白为研究对象,采用Box-Behnken响应面分析法优化绿豆蛋白水解物的制备工艺。在单因素实验的基础上,选取水解时间、底物浓度、加酶量、水解温度为影响因素,以对脱氧胆酸钠的结合能力为考察指标,采用响应面分析法得到绿豆蛋白水解物的最优制备工艺。对最优条件下制备的绿豆蛋白水解物进行超滤,随后采用Sephacryl S-100 High Resolution凝胶层析柱进行纯化得到降血脂作用较好的水解物。结果表明:在水解时间为146 min,底物浓度为3.1 g/100 mL,加酶量为9861 U/g,水解温度为52.1 ℃条件下得到的水解物与脱氧胆酸钠结合效果最好,结合率为60.46%,与模型预测值61.86%接近,误差较小。在上述最优条件下制备的绿豆蛋白降血脂水解物经超滤后,水解物的截留液呈现出更好的活性。

Preparation and Purification of Hydrolysate of Mung Bean Protein for Reducing Blood Lipids

The mung bean protein prepared by alkali extraction and acid precipitation was used to optimize the preparation process mung bean protein hydrolysate by Box-Behnken response surface methodology. Based on the results of the single factor experiments,hydrolysis time,substrate concentration,enzyme dosage and hydrolysis temperature were selected as influence factor to investigate the binding ability of mung bean protein hydrolysate with sodium deoxycholate by the response surface methodology. Ultrafiltration was carried out on mung bean protein hydrolysate prepared under the optimal conditions. Then the hydrolysate with better hypolipidemic effect was obtained through the purification by Sephacryl S-100 High Resolution gel chromatography column. The results showed that the highest combination effect of the hydrolysate and sodium deoxycholate was obtained under these conditions:Hydrolysis time was 146 min,substrate concentration was 3.1 g/100 mL,enzyme dosage was 9861 U/g,and hydrolysis temperature was 52.1 ℃. And its combination ratio was 60.46%,which was close to the predicted value of 61.86%,with little error. The retentate of the hydrolysate was much more active after ultrafiltering the mung bean protein lipid-lowering hydrolysate,which was prepared in the way mentioned above.