固定化碱性蛋白酶制备麦胚抗氧化肽条件优化  

Optimization on Preparation Conditions of Wheat Germ Antioxidant Peptides with Immobilized Alkaline Protease

作　　者：吴定[1] 孙嘉文[1] 黄卉卉[1] 谢惠惠[1] 路桂红[1] 刘长鹏[1] 

机构地区：[1]南京财经大学食品科学与工程学院/江苏省现代粮食流通与安全协同创新中心/江苏高校粮油质量安全控制及深加工重点实验室,南京210023

出　　处：《中国粮油学报》2016年第7期114-119,共6页Journal of the Chinese Cereals and Oils Association

基　　金：江苏省教育厅高新技术产业(JHB04-003)

摘　　要：利用冻干麦胚蛋白粉为原料,筛选影响固定化碱性蛋白酶制备麦胚抗氧化肽工艺条件的因素,并利用影响因素优化麦胚抗氧化肽制备工艺。采用Plackett-Burman试验设计,筛选出显著影响因素。利用Box-Behnkeny试验设计,显著影响因素进行响应曲面优化试验。结果显示,显著影响因素从大到小依次为麦胚蛋白悬浊液pH值、酶水解温度、底物浓度;响应面分析试验得到麦胚抗氧化肽对自由基清除率的回归方程,方程达到极显著水平(F=35.62,P=0.000 5),拟和很好(R^2_(Pred)=0.964 5,R^2_(Adj)=0.957)。固定化蛋白酶制备麦胚抗氧化肽优化工艺参数:底物质量浓度为4.92 g/100 mL、麦胚蛋白悬浊液的pH为9.2和酶水解温度为58.5℃;在此条件下,抗氧化肽对自由基的清除率达到55.62%。固定化蛋白酶重复使用,第18次时抗氧化肽的相对产率达到91.47%。The effect factor on preparation conditions of wheat germ antioxidant peptides with immobilized alka- line protease was screened based on using freeze - drying wheat germ dried white as raw materials and the preparation process of wheat germ antioxidant peptides was optimized by making use of effect factors. The significant influence factors were screened in the experiment design of Plackett - Burman. The optimization experimentation was done to significant influence factors based on experiment design of Box - Behnken. The results showed that effect of suspen- sion pH value of wheat germ protein was greater than that of the enzyme hydrolysis temperature and the concentration of substrate, and the effect of substrate concentration was the minimum during three influence factors. The regression equation on the clearance rate of wheat germ antioxidant peptides for free radical was obtained by the response surface analysis test, and equation reached extremely significant level （ F= 35.62, P = 0. 000 5 ） with good fitting （ Rpred2 = 0. 964 5, RZai = 0. 957 ）. The optimization process parameters of wheat germ antioxidant peptides with immobilized al- kaline protease were showed as follow ： the concentration of substratc of 4.92% , wheat germ protein suspension pH value of 9.2, and enzyme hydrolysis temperature of 58.5 ℃, on this condition the clearance rate of wheat germ an- tioxidant peptides for free radical reached 55. 62%. The relative yield of wheat germ antioxidant peptides was 91.47% at eighteenth of repeated use of the immobilized alkaline protease.

关 键 词：固定化蛋白酶 麦胚抗氧化肽 因素筛选 工艺优化 

分 类 号：TS210.9[轻工技术与工程—粮食、油脂及植物蛋白工程]