食品科学 ›› 2022, Vol. 43 ›› Issue (18): 286-291.doi: 10.7506/spkx1002-6630-20210930-362

• 安全检测 • 上一篇    

巴西坚果过敏原Ber e 1的分离纯化及表征鉴定

唐宇,徐骥远,张英,罗水忠,吴志华   

  1. (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)
  • 发布日期:2022-09-28
  • 基金资助:
    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)

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

TANG Yu, XU Jiyuan, ZHANG Ying, LUO Shuizhong, WU Zhihua   

  1. (1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China; 2. College of Food Science & Technology, Nanchang University, Nanchang 330047, China; 3. School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, China; 4. Sino German Joint Research Institute, Nanchang University, Nanchang 330047, China)
  • Published:2022-09-28

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。

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

Abstract: 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.

Key words: Brazil nut; allergen; Ber e 1; purification; anion exchange chromatography; characterization; identification

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