Effect of Heat Treatment on the Antigenicity and Conformation of Peanut Allergen Ara h 2
HU Chun-qiu1, 2, GAO Jin-yan1, 3, CHEN Hong-bing1, 2*, LUO Chun-ping1, 2, YAN Fei1, 2
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. Department of Food Science, Nanchang University, Nanchang 330047, China
摘要： 从花生种子中分离纯化花生过敏原Ara h 2，采用酶联免疫吸附试验(ELISA)、圆二色谱(CD)、ANS荧光探针及紫外可见(UV-Vis)光谱等方法，系统研究热加工对Ara h 2抗原性和结构的影响。结果表明：Ara h 2蛋白经55或70 ℃处理后其抗原性略有升高，经85，100或115 ℃处理后，其抗原性显著降低，且随着温度和时间的增加其抗原性均不断降低。CD色谱分析表明，Ara h 2经热处理后其二级结构发生变化;ANS荧光探针光谱显示，不同的热处理均导致Ara h 2表面疏水性增加。紫外光谱显示，不同温度对Ara h 2处理30 min后(除55 ℃外)，其紫外吸收值均升高。Ara h 2经100 ℃处理不同时间后，其紫外吸收值均有增加。由此推断，花生过敏原Ara h 2的构象改变导致了其抗原性的降低。
关键词：Ara h 2;热加工;抗原性;圆二色谱;ANS荧光探针;紫外光谱
Abstract：Peanut allergen Ara h 2 was extracted from peanuts and was identified by SDS-PAGE and MALDI-TOF-MS. Effect of heat treatment on the antigenicity and structure of Ara h 2 was measured by indirect ELISA, CD, fluorescence and UV absorption spectra. The results showed that the antigenicity of Ara h 2 had a slight increase after being heated at 55 or 70 ℃, while above 85 ℃ the antigenicity decreased significantly, and the antigeicity of Ara h 2 decreased with increasing temperature. The CD showed that the secondary structure of Ara h 2 was changed after heat treatment. The ANS fluorescence probe emission spectra analysis demonstrated that the heat treatment induced an increase in surface hydrophobicity of Ara h 2. The UV absorption spectra showed that the absorption maximum wavelength was increased when Ara h 2 was heated except the sample heating at 50 ℃ for 30 min. So the changes in conformation of Ara h 2 lead to the antigenicity degression.
Key words：Ara h 2;Heat treatment;Antigenicity;Circular dichroism;ANS fluorescence probe;Ultraviolet spectrum
胡纯秋1, 2，高金燕1, 3，陈红兵1, 2*，罗春萍1, 2，闫 飞1, 2 . 热加工对花生过敏原Ara h 2抗原性及构象的影响 [J]. 光谱学与光谱分析, 2010, 30(09): 2550-2554.
HU Chun-qiu1, 2, GAO Jin-yan1, 3, CHEN Hong-bing1, 2*, LUO Chun-ping1, 2, YAN Fei1, 2 . Effect of Heat Treatment on the Antigenicity and Conformation of Peanut Allergen Ara h 2 . SPECTROSCOPY AND SPECTRAL ANALYSIS, 2010, 30(09): 2550-2554.
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