食物过敏原结构变化的光谱学研究进展

摘要/Abstract

摘要： 光谱学方法已广泛应用于研究食物过敏原结构的变化。常用的光谱学方法包括圆二色光谱、荧光光谱、傅里叶变换红外光谱、拉曼光谱、紫外吸收光谱等，其中圆二色光谱仅应用于澄清的稀溶液中，常研究过敏原二级、三级结构变化；傅里叶变换红外光谱不受过敏原相对分子质量大小、光散射以及荧光的影响，其酰胺Ⅰ带对于过敏原二级结构的研究最有价值；拉曼光谱采用光子探针，对微克级样品无损伤探测，适用于稀有、珍贵样品；荧光光谱多用于研究过敏原在变性过程中疏水性以及微环境变化；紫外吸收光谱在结构鉴定中需与其他光谱方法密切配合。前3种方法均可以确定过敏原二级结构的类型和相对含量，各方法的具体应用仍然值得进一步探索。

关键词: 蛋白结构, 光谱学, 过敏原, 食物过敏

Abstract: Spectroscopy has been widely used in the investigation of structural change of food allergens. The commonly used spectroscopic techniques include circular dichroism (CD), fluorescence spectroscopy, Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy and UV absorption spectroscopy. CD is often used to define the change in secondary and tertiary structures of allergens, but is restricted to clear low-concentration solutions only. FTIR can override influence of allergen molecular weight, light scattering and fluorescent light. FTIR amide I band is the most valuable for the analysis of secondary structure of allergens. Raman spectroscopy with photon probe is suitable for non-destructive detection of rare and precious microgram-scale samples. Fluorescence spectroscopy is often used to define hydrophobic and microenvironmental changes of allergens in the course of denaturation, and UV absorption spectroscopy needs to be coordinated closely with other spectroscopic methods for structural identification. In addition, CD, fluorescence spectroscopy and FTIR are available for characterizing the type and relative content of secondary structure of food allergens, and their specific applications are still worthy of further exploration.

Key words: protein structure, spectroscopy, allergen, food allergy

中图分类号:

TS201.6

白雨鑫1,2，李欣1,2，赵丽霞3，陈红兵1,4，高金燕1,2,*. 食物过敏原结构变化的光谱学研究进展[J]. 食品科学, 2012, 33(17): 284-289.

参考文献

[1] KAGAN R S. Food allergy: an overview[J]. Environ Health Perspect, 2003, 111(2): 223–225.
[2] FERNANDEZ-RIVAS M, BOLHAAR S, GONZALEZ-MANCEBO E, et al. Apple allergy across europe: how allergen sensitization profiles determine the clinical expression of allergies to plant foods[J]. Journal of Allergy Clinical Immunol, 2006, 118(2): 481–488.
[3] GREENFIELD N, FASMAN G D. Computed circular dichroism spectra the evalution of protein conformation[J]. Biochemistry, 1969, 8(10): 4108-4116.
[4] JOHNSON W C. Analyzing protein circular dichroism spectra for accurate secondary structures[J]. Proteins Structure Function and Bioinformatics, 1999, 35(3): 307-312.
[5] DOCKAL M, CARTER D C, RUKER F. Conformational transitions of the three recombinant domains of human serum albumin depending on pH[J]. J Biol Chem, 2000, 275(5): 3042-3050.
[6] 梁毅. 结构生物学[M]. 北京: 科学出版社, 2010: 261-267.
[7] 沈星灿, 梁宏, 何锡文等. 圆二色光谱分析蛋白质构象的方法及研究进展[J]. 分析化学评述与进展, 2004, 32(3): 388-394.
[8] CRONEY J C, JAMESON D M, LEARMONTH R P. Fluorescence spectroscopy in biochemistry: Teaching basic principles with visual demonstrations[J]. Biochem Mol Biol Ed, 2001, 29(2): 60-65.
[9] CHAKRABORTY A, BASAK S. pH-induced structural transitions of caseins[J]. Journal of Photochemistry and Photobiology B: Biology, 2007, 87(3): 191-199.
[10] 梁毅. 结构生物学[M]. 北京: 科学出版社, 2010: 255-256.
[11] GASYMOV O K, GLASGOW B J. ANS fluorescence: potential to augment the identification of the external binding sites of proteins[J]. Biochimica Biophys Acta, 2007, 1774(3): 403-11.
[12] 翁诗甫. 傅里叶变换红外光谱分析[M]. 北京: 化学工业出版社, 2010: 1-2.
[13] JACKSON M, MANTSCH H H. The use and misuse of FTIR spectroscopy in the determination of protein structure[J]. Crit Rev Biochem Mol Biol, 1995, 30(2): 95-120.
[14] RUCKEBUSCH C, NEDJAR-ARROUME N, MAGAZZENI S, et al. Hydrolysis of haemoglobin surveyed by infrared spectroscopy: I. solvent effect on the secondary structure of haemoglobin[J]. Journal of Molecular Structure, 1999, 478(1-3): 185-191.
[15] 朱自莹, 顾仁敖. 中国拉曼光谱研究十年(1981~1991)[J]. 光谱学与光谱分析, 1993, 13(1): 47-82.
[16] CAREY P R. Biochemical applications of Raman and resonance Raman spectroscopies[M]. New York: Academic Press, 1982: 77.
[17] RUSO J M, GONZALEZ-PEREZ A, PRIETO G, et al. Study of the interactions between lysozyme and a fully-fluorinated surfactant in aqueous solution at different surfactant-protein ratios[J]. Int J Biol Macromol, 2003, 33(1-3): 67-73.
[18] 吴丹, 徐桂英. 光谱法研究蛋白质与表面活性剂的相互作用[J]. 物理化学学报, 2006, 22(2): 254-260.
[19] KIM D A, CORNEC M, NARSIMHAN G. Effect of thermal treatment on interfacial properties of β-lactoglobulin[J]. Journal of Colloid and Interface Science, 2005, 285(1): 100-109.
[20] BOYE J I, ISMAIL A A, ALLI I. Effects of physicochemical factors on the secondary structure of beta-lactoglobulin[J]. J Dairy Res. 1996 63(1): 97-109.
[21] LIU G M, CHENG H, NESBIT J B, et al. Effects of Boiling on the IgE-Binding Properties of Tropomyosin of Shrimp(Litopenaeus vannamei)[J]. Journal of Food Science, 2010, 75(1): T1-T5.
[22] 黄惠华, 梁汉文, 郭乾初. 超声波对大豆胰蛋白酶抑制剂活性及二级结构的影响[J]. 食品科学, 2004, 25(3): 29-33.
[23] 吴序栎, 朱倩倩, 成小娟等. 光谱法研究鸡蛋溶菌酶热变性与免疫原性的关系[J]. 分析化学研究报告, 2011, 39(2): 198-201.
[24] CAIROLI S, IAMETTI S, BONOMI F. Reversible and irreversible modifications of beta-lactoglobulin upon exposure to heat[J]. J Protein Chem, 1994, 13(3): 347-54.
[25] LALIGANT A, DUMAY E, VALENCIA C C, et al. Surface hydrophobicity and aggregation of beta-lactoglobulin heated near neutral pH[J]. J Agric Food Chem., 1991, 39 (12): 2147–2155.
[26] 张国文, 王安萍, 蒋婷等. 荧光光谱法研究橙皮苷与牛血清白蛋白相互作用特征[J]. 分析实验室, 2008, 27(1): 1-4.
[27] 涂宗财, 王辉, 刘光宪等. 动态超高压微射流对卵清蛋白微观结构的影响[J]. 光谱学与光谱分析, 2010, 30(2): 495-498.
[28] DAS K P, KINSELLA J E. Effect of heat denaturation on the adsorption of β-lactoglobulin at the oil/water interface and on coalescence stability of emulsions[J]. Journal of Colloid and Interface Science, 1990, 139(2): 551-560.
[29] CIESLA K; VANSANT E F. Physico-chemical changes taking place in gamma irradiated bovine globulins studied by thermal analysis[J]. J Therm Anal Calorim, 2010, 99(1): 315-324.
[30] 张明琦, 高美须, 支玉香等. 辐照对蟹过敏蛋白生化性质和抗原性的影响[J]. 中国农业科学, 2009, 42(9): 3259-3264.
[31] ROYCHAUDHURI R, SARATH G, ZEECE M, et al. Stability of the allergenic soybean Kunitz trypsin inhibitor[J]. Biochimica et biophysica acta-proteins and proteomics, 2004, 1699(1-2): 207-212.
[32] 陈蓁蓁, 张宁, 张文申等. 蛋白质分子荧光探针研究及其应用新进展[J]. 分析化学, 2006, 34(9): 1341-1347.
[33] SMITH D, GALAZKA V B, WELLNER N, et al. High pressure unfolding of ovalbumin[J]. International Journal of Food Science & Technology, 2000, 35(4): 361-370.
[34] NAVARRA G, LEONE M, MILITELLO V, et al. Influence of metal ions on thermal aggregation of bovine serum albumin: aggregation kinetics and structural changes[J]. J Inorq Biochem, 2009, 103(12): 1729-1738.
[35] MURAYAMA K, TOMIDA M. Heat-induced secondary structure and conformation change of bovine serum albumin investigated by Fourier transform infrared spectroscopy[J]. Biochemistry, 2004, 43(36): 11526-11532.
[36] SOMKUTI J, HOUSKA M, SMELLER L. Pressure and temperature stability of the main apple allergen Mal d 1[J]. Eur Biophys J, 2011, 40(2): 143-151.
[37] MEERSMAN F, HEREMANS K. High pressure induces the formation of aggregation-prone states of protein under reducing conditions[J]. Biophys Chem, 2003, 104(1): 297-304.
[38] ALIX A J P, PEDANDOU G, BERJOT M. Fast determination of the quantitative secondary structure of proteins by using some parameter s of the Raman amide I band[J]. Journal of Molecular Structure, 1988, 174: 159-164.
[39] HERRERO A M, JIMENEZ-COLMENERO F, CARMONA P. Elucidation of structural changes in soy protein isolate upon heating by Raman spectroscopy[J]. International Journal of Food Science and Technology, 2009, 44(4): 711-717.
[40] ASHTON L, BLANCH E W. pH-induced conformational transitions in a-lactalbumin investigated with two-dimensional Raman correlation variance plots and moving windows[J]. Journal of Molecular Structure, 2010, 974(1-3): 132-138.
[41] FERRER E G, GOMEZ A V, ANON M C, et al. Structural changes in gluten protein structure after addition of emulsi?er. A Raman spectroscopy study[J]. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 2011, 79(1): 278-281.
[42] 胡纯秋, 高金燕, 陈红兵等. 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(9): 2550-2554.
[43] LIU X H, SHANG L, JIANG X, et al. Conformational changes of ?2-lactoglobulin induced by anionic phospholipid[J]. Biophsical Chemistry, 2006, 121(3): 218-223.
[44] SU R X, QI W, HE Z M, et al. Multilevel structural nature and interactions of bovine serum albumin during heat-induced aggregation process[J]. Food Hydrocolloids, 2008, 22(6): 995-1005.
[45] RUSO J M, GONZALEZ-PEREZ A, PRIETO G, et al. Study of the interactions between lysozyme and a fully-fluorinated surfactant in aqueous solution at different surfactant-protein ratios[J]. Int J Biol Macromol, 2003, 33(1-3): 67-73.