食品科学 ›› 2010, Vol. 31 ›› Issue (15): 76-80.doi: 10.7506/spkx1002-6630-201015017

• 基础研究 • 上一篇    下一篇

花生过敏原Ara h 6的分离纯化及鉴定

罗春萍1,2,3,高金燕4,胡纯秋1,2,陈红兵1,2,*,闫 飞2,3   

  1. 1. 南昌大学 食品科学与技术国家重点实验室 2.南昌大学 中德联合研究院 3.南昌大学环境与化学工程学院 4.南昌大学生命科学与食品工程学院
  • 收稿日期:2010-04-22 出版日期:2010-08-15 发布日期:2010-12-29
  • 通讯作者: 陈红兵1 E-mail:chbgjy@hotmail.com
  • 基金资助:

    南昌大学食品科学与技术国家重点实验室目标导向项目(SKLF-MB-200807);
    江西省主要学科学术和技术带头人培养项目([2004]234 号);教育部新世纪优秀人才支持计划项目(NCET-08-07-04)

Purification and Identification of Peanut Allergen Ara h 6

LUO Chun-ping1,2,3,GAO Jin-yan4,HU Chun-qiu1,2,CHEN Hong-bing1,2,*,YAN Fei2,3   

  1. (1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China ;
    2. Sino-German Joint Research Institute, Nanchang University, Nanchang 330047, China ;
    3. School of Environmental and Chemical Engineering, Nanchang University, Nanchang 330047, China;
    4. School of Life Sciences and Food Engineering, Nanchang University, Nanchang 330047, China)
  • Received:2010-04-22 Online:2010-08-15 Published:2010-12-29
  • Contact: CHEN Hong-bing1 E-mail:chbgjy@hotmail.com

摘要:

为高效分离纯化花生过敏原Ara h 6,通过脱脂、蛋白浸提、阴离子交换层析分离得到目的蛋白,并用十二烷基磺酸钠- 聚丙烯酰胺凝胶电泳(SDS-PAGE)、基质辅助激光解吸/ 电离飞行时间质谱(MALDI-TOF/MS)及免疫印迹技术(Western blotting)对其进行鉴定。结果表明,该蛋白为花生过敏原Ara h 6,其分子质量约为15kD,纯度大于95%,得率为22.5%。该方法简单、高效,可为花生过敏的进一步研究提供实验材料。

关键词: 花生过敏, Ara h 6, 分离纯化

Abstract:

In order to effectively isolate and purify peanut allergen Ara h 6, peanut was subjected to a series of sequential treatments, namely de-fatting, protein extraction and anion-exchange chromatographic. Sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE), matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF/MS) and Western blotting were used for the identification of target protein. The results indicated that the purified target protein was peanut allergen Ara h 6 with a molecular weight of 15 kD. The purity of Ara h 6 was more than 95%, and the recovery rate was 22.5%. Based on these investigations, a simple and efficient approach to purifying Ara h 6 is achieved, which will provide experimental materials for further research on peanut allergy.

Key words: peanut allergy, Ara h 6, isolation and purification

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