碧根果致敏原Car i 1的分离纯化及表征鉴定  

Separation,purification,identification and characterization of major allergen Car i 1 from Carya illinoensis

作　　者：罗祺书 唐宇[1,2] 张英 朱伟超[1,2] 赵凯文 罗水忠 吴志华 LUO Qi-Shu;TANG Yu;ZHANG Ying;ZHU Wei-Chao;ZHAO Kai-Wen;LUO Shui-Zhong;WU Zhi-Hua(State Key Laboratory of Food Science and Resources,Nanchang University,Nanchang 330047,China;School of Food Science&Technology,Nanchang University,Nanchang 330031,China;Nanchang Institute of Technology Medical College,Nanchang 330044,China;School of Food and Biological Engineering,Hefei University of Technology,Hefei 230009,China;Sino-German Joint Research Institute,Nanchang University,Nanchang 330047,China)

机构地区：[1]南昌大学食品科学与资源挖掘全国重点实验室,南昌330047 [2]南昌大学食品学院,南昌330031 [3]南昌理工学院医学院,南昌330044 [4]合肥工业大学食品与生物工程学院,合肥230009 [5]南昌大学中德联合研究院,南昌330031

出　　处：《食品安全质量检测学报》2024年第4期13-20,共8页Journal of Food Safety and Quality

基　　金：国家重点研发计划项目(2023YFF1104002);国家自然科学基金项目(32160540);安徽省科技重大专项(202003a06020025);江西省教育厅项目(GJJ212121)。

摘　　要：目的分离纯化碧根果致敏原Car i 1,并对其结构进行表征鉴定。方法以新鲜碧根果果仁为原料,通过粉碎、脱脂、浸提、粗分级、凝胶过滤层析,对碧根果致敏原蛋白Car i 1进行分离纯化。结合十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、液相色谱-串联质谱法和免疫印迹法3种方法对Cari1进行鉴定,并通过圆二色谱仪与紫外分光光度计表征其二、三级结构。结果本方法纯化获得碧根果致敏原Cari1,单轮制备量可达5 mg以上,且纯度大于95%,蛋白质高级结构未被破坏,能够被全部3名碧根果过敏患者的血清准确识别。结论该纯化方法技术路线简单、设备要求低且单次制备量高,总得率可达65%,操作便捷,为碧根果致敏原Car i 1的相关研究奠定了物质基础。Objective To isolate and purify the allergen Car i 1 from Carya illinoinensis,and characterize and identify its structure.Methods Taking fresh Carya illinoensis as raw material,the allergen protein Car i 1 of Carya illinoensis was isolated and purified by crushing,degreasing,extraction,rough grading and gel filtration chromatography.Car i 1 was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis,liquid chromatography-tandem mass spectrometry and immunoblotting,and its secondary and tertiary structures were characterized by circular dichroism and ultraviolet spectrophotometer.Results This method purified the allergen Car i 1 from Carya illinoensis,with a single round preparation amount of over 5 mg and a purity of over 95%.The protein's advanced structure was not damaged and could be accurately recognized by the serum of all 3 Carya illinoensis allergy patients.Conclusion The purification method has the advantages of simple technical route,low equipment requirements,high single preparation quantity,total rate up to 65%,and convenient operation,which lays a material foundation for the related research of allergen Car i 1 from Carya illinoinensis.

关 键 词：碧根果 致敏原 Car i 1 分离纯化 凝胶过滤层析 

分 类 号：TS201.4[轻工技术与工程—食品科学]