花生蛋白Ara h1分子结构变化及其对致敏活性的影响

项目名称： 花生蛋白Ara h1分子结构变化及其对致敏活性的影响

项目编号： No.31471614

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 薛文通

作者单位： 中国农业大学

项目金额： 89万元

中文摘要： 花生蛋白是一种理想的食品加工原料，同时也是一种致死率较高的食物过敏原，严重危害人类健康。本课题以花生蛋白中主要的过敏原Ara h1为研究对象，采用热处理、超高压及糖基化等不同改性加工方法对其进行处理，研究改性前后蛋白分子组成及构象的变化、体内体外肥大细胞脱颗粒过程致敏性的变化以及两者之间的构效关系，揭示改性对花生蛋白Ara h1致敏性的影响机制。在分子结构上，拟利用圆二色光谱和核磁共振分析改性前后蛋白结构的变化，模拟胃肠消化分析致敏肽段分子组成；在细胞层面上，利用ELISA法考察各介质分泌性的变化，同位素掺入法和CKK-8法检测细胞增殖能力变化，实时荧光定量PCR法检测转录水平基因表达量变化；在动物层面上，采用电镜等方法观察组织形态和病理学变化。从而在体内、体外以及分子水平综合揭示各种改性加工方法对花生Ara h1致敏性的影响规律，为今后开发安全有效的脱敏方法奠定理论基础。

中文关键词： 花生过敏原Ara；h1；改性加工；致敏活性；蛋白结构；构效关系

英文摘要： Peanut protein is an ideal resource for food processing, however it is also an important food allergen seriously threatening human health, with the highest food allergy lethality rate. This project takes Ara h1, the major peanut allergen in China, as the research object, and treats it with diverse modification processing approaches, including heat treatment, hyper-pressure, and glycosylation. Regulatory mechanisms of modification processing on sensitization of peanut protein Ara h1 will be explored based on measuring the changes of protein compositions and conformations, sensitizations of in vivo and in vitro mastocytes degranulation processes, and the structure-function relationship between the above two factors. On the level of molecular structure, protein conformation changes before and after modifications will be analyzed by circular dichroism and NMR, molecular compositions of the allergic peptides will be analyzed by simulation of gastrointestinal digestion. On the cellular level, changes of medium secretion will be detected by ELISA, changes of cell proliferation capacities will be checked by the methods of isotope- incorporation and CKK-8 method, gene expression changes on transcription level will be calculated by RFFQ- PCR. On the animal level, organization form and pathological changes will be observed by electron microscope. In summary, desensitization mechanisms of diverse approaches will be systematically disclosed on in vitro, in vivo, and molecular levels, so as to provide theoretical basis for the development of desensitivition methods for peanuts in future.

英文关键词： peanut allergen Ara h1;modification;Sensitization;protein structure;structure-function relationship