巴西坚果过敏原Ber e 1的分离纯化及表征鉴定  

Purification,Identification and Characterization of Brazil Nut Allergen Ber e 1

作　　者：唐宇 徐骥远 张英[1,2] 罗水忠[3] 吴志华[1,4] TANG Yu;XU Jiyuan;ZHANG Ying;LUO Shuizhong;WU Zhihua(State Key Laboratory of Food Science and Technology,Nanchang University,Nanchang 330047,China;College of Food Science&Technology,Nanchang University,Nanchang 330047,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技术国家重点实验室,江西南昌330047 [2]南昌大学食品学院,江西南昌330047 [3]合肥工业大学食品与生物工程学院,安徽合肥230009 [4]南昌大学中德联合研究院,江西南昌330047

出　　处：《食品科学》2022年第18期286-291,共6页Food Science

基　　金：国家自然科学基金地区科学基金项目(32160540);江西省自然科学基金重点项目(20212ACB205013);安徽省科技重大专项(202003a06020025)。

摘　　要：以巴西坚果仁为原料,通过粉碎、脱脂、浸提、阴离子交换层析纯化过敏原蛋白Ber e 1;利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、液相色谱-串联质谱联用、Western blot等方法对其进行鉴定,并通过圆二光谱仪和紫外分光光度计表征其二、三级结构。结果表明:纯化获得的巴西坚果过敏原Ber e 1,单轮制备量可达20 mg以上,且纯度大于95%,其蛋白质高级结构未被破坏,能够被巴西坚果过敏患者的血清准确识别。该纯化技术路线简单、对设备要求低且高效,为巴西坚果过敏原Ber e 1的相关研究奠定了一定的基础。The allergen protein Ber e 1 was purified from Brazil nuts by crushing,defatting,extraction,and anion exchange chromatography,and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE),liquid chromatography-tandem mass spectrometry(LC-MS/MS)and Western blot,and its secondary and tertiary structures were characterized by circular dichroism(CD)and UV spectroscopy.The results showed that the purified target protein was identified as Ber e 1,and the yield of Ber e 1 was more than 20 mg per round,and its purity was higher than 95%.The purified allergen showed undamaged secondary or tertiary structures,and could be recognized by the serum of allergic patients.This purification method is simple and efficient with low instrument requirements,which can provide a basis for Ber e 1 research.

关 键 词：巴西坚果 过敏原 Ber e 1 纯化 阴离子交换层析 表征 鉴定 

分 类 号：TS255.1[轻工技术与工程—农产品加工及贮藏工程]