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.
[1] Shewry P R, Napier J A, Tatham A S. Plant Cell, 1995, 7(7): 945. [2] Sampson H A. Engl. N. J. Med., 2002, 346(17): 1294. [3] Al-Muhsen S, Clarke A E, Kagan R S. CMAJ, 2003, 168(10): 1279. [4] Stanley J S, King N, Burks A W, et al. Arch. Biochem. Biophys., 1997, 342(2): 244. [5] Palmer G W, Dibbern D A Jr, Burks A W, et al. Clin. Immunol., 2005, 115(3): 302. [6] Schappi G F, Konrad V, Imhof D, et al. Allergy, 2001, 56(12): 1216. [7] Sathe S K, Teuber S S, Roux K H. Biotechnol. Adv., 2005, 23(6): 423. [8] HU Chun-qiu, GAO Jin-yan, CHEN Hong-bing(胡纯秋, 高金燕, 陈红兵) . Food Science(食品科学), 2006, 27(12): 784. [9] Beyer K, Morrow E, Li X M, et al. J. Allergy Clin. Immunol., 2001, 107(6): 1077. [10] Mondoulet L, Paty E, Drumare M F, et al. J. Agric. Food Chem., 2005, 53(11): 4547. [11] Laemmli U K. Nature, 1970, 227: 680. [12] Lowry O H, Rosebrough N J, Farr A L, et al. J. Biol. Chem., 1951, 193: 265. [13] Takagi S, Akashi M, Yasumatsu K. Nippon Shokuhin Kogyo Gakkaishi, 1979, 26: 133. [14] Koppelman S J, Wensing M, Ertmann M, et al. Clin. Exp. Allergy, 2004, 34(4): 583. [15] Bu G H, Lou Y K, Zheng Z, et al. Food Agr. Immunol., 2009, 20(3): 195. [16] DONG Xiao-wei, SONG Xiao-yan , SHI Mei, et al(董小卫,宋晓妍,石 梅, 等). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2010, 30(2): 458. [17] Pasdar N A, Li-Chan E C Y. J. Agric. Food Chem., 2000, 48(2): 328. [18] WU Dan, XU Gui-ying(吴 丹, 徐桂英). Acta Phys-Chim. Sin.(物理化学学报), 2006, 22(2): 254.