酶解改性对小麦醇溶蛋白致敏性的影响与工艺优化  

Effect of Enzymatic Hydrolysis Modification on the Allergenicity of Wheat Gliadin and Process Optimization

作　　者：孟智鹏 温青玉 张雨 李天齐 张康逸 MENG Zhipeng;WEN Qingyu;ZHANG Yu;LI Tianqi;ZHANG Kangyi(Institute of International Education,Henan University of Technology,Zhengzhou 450007,China;Center of Agricultural Products Processing,Henan Academy of Agricultural Sciences/Henan Province International Joint Laboratory for Whole Grain Wheat Processing/Henan Province Whole Grain Fresh Food Processing Engineering Technology Research Center,Zhengzhou 450002,China;Henan Ankang Food Science and Technology Research Institute,Zhengzhou 450047,China;Henan Ankang Future Food Technology Co.,Ltd.,Zhengzhou 450066,China)

机构地区：[1]河南工业大学国际教育学院,河南郑州450007 [2]河南省农业科学院农副产品加工研究中心/河南省全谷物小麦制品加工国际联合实验室/河南省全谷物鲜食加工工程技术研究中心,河南郑州450002 [3]河南省安康食品科技研究院,河南郑州450047 [4]河南省安康未来食品科技有限公司,河南郑州450066

出　　处：《河南农业科学》2022年第7期163-172,共10页Journal of Henan Agricultural Sciences

基　　金：河南省重大公益专项(201300110300);河南省科技攻关项目(222102110256);河南省农业科学院优秀青年科技基金项目(2022YQ30);河南省农业科学院科技创新团队项目(2022TD24)。

摘　　要：小麦醇溶蛋白(GLI)是小麦中的主要过敏原,为降低其致敏性,研究酶法改性对 GLI 致敏性的影响,并对酶解工艺进行优化。首先,以致敏性特征值OD_(450)为评价指标,筛选出碱性蛋白酶和复合蛋白酶进行同步酶解。然后以 OD_(450)和水解度为指标进行单因素试验,并通过响应面试验优化酶解工艺,得到降低 GLI 致敏性的最佳酶解工艺:酶添加量为 3 080 U/g,底物质量浓度为 32 g/L,酶解温度为 50.5 ℃,酶解 pH 值为 7.48,酶解时间为 4 h。在此条件下,OD_(450)由未处理的 1.013 2 降至 0.363 5。SDS-PAGE 和Tricine-SDS-PAGE 电泳分析显示,双酶酶解比单酶酶解有更高的酶解效率。ELISA 和 Western blot分析显示,酶法改性降低GLI致敏性的效果明显,且最优酶配方(双酶)比单酶降低致敏性的效果更佳。Wheat gliadin(GLI)is the main allergen in wheat.The effect of enzymatic modification on the sensitization of GLI was studied,and the enzymatic hydrolysis process was optimized to reduce its sensitization.First,alkaline protease and complex protease were screened for simultaneous enzymatic hydrolysis with the sensitization characteristic value OD_(450) as the evaluation index.Then a single factor experiment was carried out with the OD_(450) and the degree of hydrolysis as indexes.The enzymatic hydrolysis process was optimized by response surface methodology,and the optimal enzymatic hydrolysis process for reducing the sensitization of GLI was obtained as follows:The enzyme addition amount was3 080 U/g,the substrate mass concentration was 32 g/L,the enzymatic hydrolysis temperature was50.5 ℃,the pH value was 7.48,and the time was 4 h.Under this condition,the OD_(450) value decreased from 1.013 2 to 0.363 5.SDS-PAGE and Tricine-SDS-PAGE analyses showed that double-enzyme hydrolysis had higher enzymatic hydrolysis efficiency than single-enzyme hydrolysis.ELISA and Western blot analyses showed that the effect of enzymatic modification in reducing the sensitization of GLI was good,and the optimal enzyme formula(double enzyme)was better than the single enzyme in reducing sensitization.

关 键 词：小麦 醇溶蛋白 致敏性 酶解 工艺优化 响应面 

分 类 号：S512.1[农业科学—作物学] TS210.4[轻工技术与工程—粮食、油脂及植物蛋白工程]