微波处理条件对蛋清结构和IgG结合能力的影响
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摘要
为探究降低蛋清致敏性的新方法,在不同的微波条件下处理蛋清蛋白,通过SDS-PAGE(十二烷基磺酸钠-聚丙烯酰氨凝胶电泳)分析分子量变化、圆二色光谱分析二级结构变化、紫外光谱分析三级结构变化、外源性荧光光谱分析疏水性变化、间接ELISA(酶联免疫吸附试验)分析IgG(免疫球蛋白G)结合能力的变化,研究微波对蛋清的影响。研究证实,微波处理降低蛋清蛋白的潜在致敏性,其中400 W/30 s处理后的蛋白显示出最低的IgG结合能力,所有条件下处理后的蛋清蛋白的二、三级结构均发生显著改变。80 W的微波功率下,蛋白质的结构先折叠,该条件下处理30 s后,蛋白结构又展开;当微波功率达到640 W时,处理时间较长的蛋白分子产生堆叠。蛋白质分子的结构变化导致了其抗原性的变化。
Aiming to explore a new way to reduce the allergenicity of egg allergens. The influence of different microwave treatments on egg white was demonstrated by SDS-PAGE, circular dichroism spectroscopy,ultraviolet spectroscopy, exogenous fluorescence spectroscopy and indirect ELISA to analyze molecular weight changes,secondary structure,tertiary structure,hydrophobic changes and changes in IgG binding capacity,respectively. Results showed that the microwave treatment reduced the potential allergenicity of egg white proteins,and the IgG binding capacity of egg white was the lowest when it was treated under 400 W for 30 s. The secondary and tertiary structure of protein were obviously changed by microwave treatment. Under 80 W,the structure of egg white proteins was folded,however,when treatment time was more than 30 s,the structure of egg white proteins was unfolded again. When the power of microwave treatment was increased to 640 W,part of egg white proteins was aggregated.Changes in the structure could induce changes in potential allergenicity of egg white proteins,however,the relationship of structure and potential allergenicity of egg white proteins under microwave treatment should be further analyzed.
Aiming to explore a new way to reduce the allergenicity of egg allergens. The influence of different microwave treatments on egg white was demonstrated by SDS-PAGE, circular dichroism spectroscopy,ultraviolet spectroscopy, exogenous fluorescence spectroscopy and indirect ELISA to analyze molecular weight changes,secondary structure,tertiary structure,hydrophobic changes and changes in IgG binding capacity,respectively. Results showed that the microwave treatment reduced the potential allergenicity of egg white proteins,and the IgG binding capacity of egg white was the lowest when it was treated under 400 W for 30 s. The secondary and tertiary structure of protein were obviously changed by microwave treatment. Under 80 W,the structure of egg white proteins was folded,however,when treatment time was more than 30 s,the structure of egg white proteins was unfolded again. When the power of microwave treatment was increased to 640 W,part of egg white proteins was aggregated.Changes in the structure could induce changes in potential allergenicity of egg white proteins,however,the relationship of structure and potential allergenicity of egg white proteins under microwave treatment should be further analyzed.
引文
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[22] ZHANG T,Lü C,YUN S,et al. Effect of high hydrostatic pressure(HHP)on structure and activity of phytoferritin[J].Food Chemistry,2012,130(2):273-278.
[2] SZéPFALUSI Z,EBNER C,PANDJAITAN R,et al. Egg yolkα-livetin(chicken serum albumin)is a cross-reactive allergen in the bird-egg syndrome[J]. J. Allergy Clin. Immunol.,1994,93(5):932-942.
[3] QUIRCE S,MARA?óN F,UMPIéRREZ A,et al. Chicken serum albumin(Gal d 5*)is a partially heat-labile inhalant and food allergen implicated in the bird-egg syndrome[J]. Allergy,2015,56(8):754-762.
[4] POZAR D M. Microwave engineering education:from field theory to circuit theory[C]∥2012 IEEE/MTT-S International Microwave Symposium Digest. IEEE,2012:1-3.
[5] JORDENS K,WILKES G L,JANZEN J,et al. The influence of molecular weight and thermal history on the thermal,rheological,and mechanical properties of metallocene-catalyzed linear polyethylenes[J]. Polymer,2000,41(19):7175-7192.
[6] MUDGETT R E. Microwave food processing[J]. Food Technology,1989,52(1):243-261.
[7] AYDOGDU A,SUMNU G,SAHIN S. Effects of microwave-infrared combination drying on quality of eggplants[J]. Food&Bioprocess Technology,2015,8(6):1198-1210.
[8] XU D,MIN Z,MUJUMDAR A S,et al. Microwave freeze drying of sea cucumber(Stichopus japonicus)[J]. Journal of Food Engineering,2010,96(4):491-497.
[9] HORUZ E,MASKAN M. Hot air and microwave drying of pomegranate(Punica granatum L.)arils[J]. Journal of Food Science&Technology,2015,52(1):285-293.
[10] GUO Q,SUN D W,CHENG J H,et al. Microwave processing techniques and their recent applications in the food industry[J]. Trends in Food Science&Technology,2017,67(1):236-247.
[11] MESSIA M C,FALCO T D,PANFILI G,et al. Rapid determination of collagen in meat-based foods by microwave hydrolysis of proteins and HPAEC-PAD analysis of 4-hydroxyproline[J]. Meat Science,2008,80(2):401-409.
[12] LANDER T A,DADONAITE B,MONRO A K. Microwave drying of plant material for herbarium specimens and genetic analysis[J]. Taxon,2013,62(4):790-797.
[13] ISMAEL R,VITTORIA B,ALEJANDROJ M. Microwave-assisted modification of starch for compatibilizing LLDPE/starch blends[J]. Carbohydrate Polymers,2009,75(2):343-350.
[14]董晓颖,高美须,潘家荣,等.不同处理方法对虾过敏蛋白分子量及抗原性的影响[J].核农学报,2010,24(3):548-554.DONG Xiaoying,GAO Meixu,PAN Jiarong,et al. Effects of different treatments on molecular weight and antigenicity of shrimp allergenic protein[J]. Journal of Nuclear Agricultural Sciences,2010,24(3):548-554.(in Chinese)
[15]刘光明,余惠琳,黄秀秀,等.加工处理方式对蟹类原肌球蛋白的消化稳定性和过敏原性的影响[J].中国食品学报,2011,11(4):14-22.LIU Guangming,YU Huilin,HUANG Xiuxiu,et al. Effect of processing method on digestive stability and allergenicity of crab tropomyosin[J]. Journal of Chinese Institute of Food Science and Technology,2011,11(4):14-22.(in Chinese)
[16]卞蓉霞.微波辅助酶解乳清蛋白的研究[D].无锡:江南大学,2013.BIAN Rongxia. Microwave-assisted proteolysis of whey proteins[D]. Wuxi:Jiangnan University,2013.(in Chinese)
[17] ACHOURI A,BOYE J I. Thermal processing,salt and high pressure treatment effects on molecular structure and antigenicity of sesame protein isolate[J]. Food Research International,2013,53(1):240-251.
[18]邢绍平.微波处理对乳清蛋白凝胶影响的探讨[J].中国乳业,2016(5):69-73.XING Shaoping. The impact of microwave treatment on whey protein gel[J]. China Dairy,2016(5):69-73.(in Chinese)
[19]林克椿.生物物理技术:波谱技术及其在生物学中的应用[M].北京:高等教育出版社,1989.
[20] PELTON J T,MCLEAN L R. Spectroscopic methods for analysis of protein secondary structure[J]. Analytical Biochemistry,2000,277(2):167-176.
[21]肖厚荣,施春华,夏炳乐,等. Cu2+与烟草多酚氧化酶相互作用研究[J].无机化学学报,2003,19(6):589-593.XIAO Hourong,SHI Chunhua,XIA Bingle,et al. Study on interaction between Cu2+and tobacco polyphenol oxidase[J].Chinese Journal of Inorganic Chemistry,2003,19(6):589-593.(in Chinese)
[22] ZHANG T,Lü C,YUN S,et al. Effect of high hydrostatic pressure(HHP)on structure and activity of phytoferritin[J].Food Chemistry,2012,130(2):273-278.