酶解制备苦荞蛋白抗氧化肽及其分离纯化研究  

Preparation and Purification of Antioxidant Peptides from Tartary Buckwheat Protein by Enzymatic Hydrolysis

作　　者：陈金玉 曲金萍 张坤生[1] 闫怡君 任云霞[1] CHEN Jin-yu;QU Jin-ping;ZHANG Kun-sheng;YAN Yi-jun;REN Yun-xia(Tianjin Key Laboratory of Food and Biotechnology,College of Biotechnology and Food Science,Tianjin University of Commerce,Tianjin 300134,China)

机构地区：[1]天津市食品生物技术重点实验室,天津商业大学生物技术与食品科学学院,天津300134

出　　处：《食品研究与开发》2020年第12期14-20,共7页Food Research and Development

基　　金：国家自然科学基金项目(31701612,31671873);“十三五”国家重点研发计划重点专项(2016YFD0401503);国家级大学生创新创业训练计划项目(201910069001)。

摘　　要：以苦荞麦粉为原料,提取苦荞蛋白,分别采用碱性蛋白酶、胃蛋白酶、胰蛋白酶对蛋白进行酶解,采用DPPH法比较不同酶解产物的抗氧化活性,从而筛选水解制备苦荞蛋白抗氧化肽的最适酶。以水解度为指标,利用单因素试验和响应面法优化酶解工艺条件。结果表明,不同蛋白酶酶解产物的抗氧化活性大小为:胃蛋白酶>胰蛋白酶>碱性蛋白酶,其中胃蛋白酶酶解产物的DPPH自由基清除率最高,为68.47%。胃蛋白酶最佳水解工艺条件为:时间2.5 h、温度38℃、pH 2.0,在此条件下苦荞蛋白水解度为32.68%。采用超滤对苦荞蛋白水解物进行分离纯化,结果表明,分子量<3 kDa的水解物具有显著的抗氧化活性;经凝胶过滤色谱进一步分离得到3个峰,小分子量峰组分显示出最强的抗氧化活性。Tartary buckwheat flour was used as raw materials to extract tartary buckwheat protein.The proteins were hydrolyzed by alkaline protease,pepsin and trypsin,and the antioxidant activities of different enzymatic products were compared by DPPH assay,so as to screen the optimal enzyme for preparing antioxidant peptides from tartary buckwheat protein by hydrolysis.The hydrolysis conditions were optimized by single factor test and response surface method.Results showed that the antioxidant activities of different enzymatic products was:pepsin>trypsin>alkaline protease,and the DPPH free radical scavenging rate of pepsin enzymatic products was 68.47%.The optimum hydrolysis conditions of pepsin were:time 2.5 h,temperature 38℃,and pH 2.0.Under these conditions,the degree of hydrolysis reached 32.68%.Ultrafiltration was used to separate and purify tartary buckwheat protein hydrolysates.Results indicated that the hydrolysates with molecular weight<3 kDa had significant antioxidant activity.On this basis,three elution peaks were further isolated by gel filtration chromatography,and the peak components with the smallest molecular weight showed the strongest antioxidant activity.

关 键 词：苦荞蛋白 抗氧化肽 水解度 超滤 凝胶过滤 

分 类 号：TQ936.16[化学工程]