紫薯多酚蛋白复合物酶解及降血糖组分制备工艺优化  

Preparation and Activity Study of Purple Sweet Potato Hypoglycemic Peptide

作　　者：李佳[1] 帅晓艳 李漫 杨宁[1] 李书艺[1] 张瑞[1] 何静仁[1] LI Jia;SHUAI Xiaoyan;LI Man;YANG Ning;LI Shuyi;ZHANG Rui;HE Jingren(Wuhan Polytechnic University,Wuhan 430023,China)

机构地区：[1]武汉轻工大学,湖北武汉430023

出　　处：《食品科技》2020年第9期43-50,共8页Food Science and Technology

基　　金：湖北省技术创新专项重大项目(2019ABA113);湖北省科技支撑计划项目(2015BHE015)。

摘　　要：采用双酶水解法结合超滤膜分离从紫薯多酚蛋白复合物中制备降血糖肽组分。以酶解产物的水解度及α-葡萄糖苷酶抑制率为指标,从木瓜蛋白酶、中性蛋白酶、碱性蛋白酶、胰蛋白酶和胃蛋白酶中筛选出2种蛋白酶,以酶解时间、pH、加酶量和酶解温度分别进行单因素和正交实验优化酶解工艺。酶解工艺最佳条件为:底物浓度0.01 g/mL,木瓜蛋白酶在pH7.0、温度50℃、酶添加量5000 U/g的条件下酶解1 h;灭活冷却后,碱性蛋白酶在pH8.0、温度60℃、酶添加量6000 U/g的条件下酶解5 h。得到酶解产物的水解度为25.72%,α-葡萄糖苷酶抑制率为24.18%。为探究酶解产物中不同分子质量的降血糖活性组分,酶解液分别通过3ku和10ku超滤膜后得到3个组分(组分1:分子质量<3 ku,组分2:分子质量3~10 ku,组分3:分子质量>10ku),通过胰岛素抵抗模型HepG2细胞实验验证3个组分的降血糖活性。结果表明,组分1可增加胰岛素抵抗模型细胞对葡萄糖的消耗量,可作为分离纯化制备紫薯高活性降血糖肽类物质的优选来源。A bi-enzyme hydrolysis method was used to prepare hypoglycemic peptide from the purple sweet potato polyphenol-protein complexes.Two enzymes with higher glucosidase inhibition,were screened from papain,neutral protease,alcalase,trypsin and pepsin.The various factors including the enzymolysis time,pH,the amount of enzyme added and the temperature of the enzymolysis process were studied in order to determine the optimal process conditions through single factor experiment and orthogonal experiment.The optimal conditions were as follows:hydrolysis with papain at pH 7.0,50℃,5000 U/g enzyme dosage for 1 h,followed by hydrolysis with alcalase at pH 8.0,60℃,6000 U/g enzyme dosage for 5 h.Under these conditions,the degree of hydrolysis of the hydrolyzed product was 25.72%,and the inhibition rate ofα-glucosidase was 24.18%.Three groups(group 1:Mw<3 ku,group 2:Mw 3~10 ku,group 3:Mw>10 ku)were further separated from the hydrolyzed product through the 3 ku and 10 ku ultrafiltration membranes.The hypoglycemic activity of the three groups was verified by the HepG2 cell experiment.The results showed that group 1 can effectively increase the glucose consumption of HepG2 cells in the insulin resistance model,which can be used as a promising source for further separation and purification of purple sweet potato hypoglycemic peptides.

关 键 词：紫薯多酚蛋白复合物 双酶水解 超滤膜分离 降血糖组分 工艺优化 

分 类 号：TS201.2[轻工技术与工程—食品科学]