食品与发酵工业 >

2023 , Vol. 49 >Issue 12: 215 - 220

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.032015

研究报告

金鲳鱼小清蛋白的纯化及其结构与性质分析

  • 邱惠 ,
  • 王新 ,
  • 段伟文 ,
  • 魏帅 ,
  • 孙钦秀 ,
  • 夏秋瑜 ,
  • 王泽富 ,
  • 韩宗元 ,
  • 刘书成
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  • 1(广东海洋大学 食品科技学院,广东省水产品加工与安全重点实验室,广东省海洋生物制品工程实验室,广东省海洋食品工程技术研究中心,水产品深加工广东普通高等学校重点实验室,广东 湛江,524088)
    2(南方海洋科学与工程广东省实验室(湛江),广东 湛江,524025)
    3(大连工业大学,海洋食品精深加工关键技术省部共建协同创新中心,辽宁 大连,116034)
硕士研究生(魏帅副教授和刘书成教授为共同通信作者,E-mail:weishuaiws@126.com;Lsc771017@163.com)

收稿日期: 2022-04-17

  修回日期: 2022-05-26

  网络出版日期: 2023-07-13

基金资助

南方海洋科学与工程广东省实验室(湛江)(ZJW-2019-06);广东普通高等学校海洋食品绿色加工技术研究团队(2019KCXTD011);广东海洋大学博士科研启动项目(R20048)

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Purification, structure, and property analysis of parvalbumin from Trachinotus ovatus

  • QIU Hui ,
  • WANG Xin ,
  • DUAN Weiwen ,
  • WEI Shuai ,
  • SUN Qinxiu ,
  • XIA Qiuyu ,
  • WANG Zefu ,
  • HAN Zongyuan ,
  • LIU Shucheng
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  • 1(College of Food Science and Technology, Guangdong Ocean University, Guangdong Provincial Key Laboratory of Aquatic Products Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of seafood, Key Laboratory of Advanced Processing of Aquatic Product of Guangdong Higher Education Institution, Zhanjiang 524088, China)
    2(Southern Marine Science and Engineering Guangdong Laboratory, Zhanjiang 524025, China)
    3(Dalian Polytechnic University, Collaborative Innovation Center of Seafood Deep Processing, Dalian 116034, China)

Received date: 2022-04-17

  Revised date: 2022-05-26

  Online published: 2023-07-13

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摘要

分离纯化金鲳鱼小清蛋白(parvalbumin, PV),并对其进行结构与性质分析。经Tris-HCl粗提、热处理、联合2次柱层析可从金鲳鱼肌肉中得到较高纯度的目的蛋白,经SDS-PAGE分析,其分子质量约10 kDa。蛋白免疫印迹显示,10 kDa处有阳性反应,进一步质谱鉴定确定其为金鲳鱼PV蛋白。PV蛋白结构分析表明,其为典型的α-螺旋结构,在248 nm处有紫外特征吸收峰,在350 nm处有内源荧光最大发射峰。PV蛋白性质分析表明,金鲳鱼PV蛋白热稳定较好,耐弱酸性、耐碱性。消化稳定性试验表明,胃蛋白酶可分解金鲳鱼PV蛋白,而金鲳鱼PV蛋白对胰蛋白酶具有抗性。

关键词： 金鲳鱼; 过敏源; 小清蛋白; 结构; 稳定性

本文引用格式

邱惠 , 王新 , 段伟文 , 魏帅 , 孙钦秀 , 夏秋瑜 , 王泽富 , 韩宗元 , 刘书成 . 金鲳鱼小清蛋白的纯化及其结构与性质分析[J]. 食品与发酵工业, 2023 , 49(12) : 215 -220 . DOI: 10.13995/j.cnki.11-1802/ts.032015

Abstract

This study aimed to isolate and purify parvalbumin (PV) from Trachinotus ovatus.The structure and property of PV were also analyzed.The protein was obtained from the Trachinotus ovatus by Tris-HCl extraction, heat treatment, and twice of column chromatography.The SDS-PAGE result showed the molecular weight of the protein was about 10 kDa.The result of the Western blot showed that positive reaction at 10 kDa, which was further identified as PV by mass spectrometry.The structure analysis of PV showed that it contained typical α-helix.The UV absorption peak at 248 nm was observed and the peak of intrinsic fluorescence was found at 350 nm.The characterization analysis of PV showed that it had thermal stability and was resistant to weak acids and alkalis.The digestion test showed that pepsin could decompose PV, but it was resistant under trypsin.

Key words： Trachinotus ovatus; allergen; parvalbumin; structure; stability

参考文献

[1] 傅玲琳, 王彦波.食物过敏:从致敏机理到控制策略[J].食品科学, 2021, 42(19):1-19.
FU L L,WANG Y B.Food allergy:From sensitization mechanism to control strategy[J].Food Science, 2021, 42(19):1-19.
[2] LIANG J, TAYLOR S L, BAUMERT J, et al.Effects of thermal treatment on the immunoreactivity and quantification of parvalbumin from Southern hemisphere fish species with two anti-parvalbumin antibodies[J].Food Control, 2021, 121:107675.
[3] GALAND C, LEYVA-CASTILLO J M, YOON J, et al.IL-33 promotes food anaphylaxis in epicutaneously sensitized mice by targeting mast cells[J].Journal of Allergy and Clinical Immunology, 2016, 138(5):1 356-1 366.
[4] HEIZMANN C W.Ca2+-Binding Proteins of the EF-Hand Superfamily:Diagnostic and Prognostic Biomarkers and Novel Therapeutic Targets[M].Calcium-Binding Proteins of the EF-Hand Superfamily.New York:Humana Press, 2019:157-186.
[5] SWOBODA I, BALIC N, KLUG C, et al.A general strategy for the generation of hypoallergenic molecules for the immunotherapy of fish allergy[J].The Journal of Allergy and Clinical Immunology, 2013, 132(4):979-981.
[6] SAPTARSHI S R, SHARP M F, KAMATH S D, et al.Antibody reactivity to the major fish allergen parvalbumin is determined by isoforms and impact of thermal processing[J].Food Chemistry, 2014, 148: 321-328.
[7] 魏帅, 刘媛, 刘蒙娜, 等.凡纳滨对虾多酚氧化酶的纯化鉴定与生物信息学分析[J].广东海洋大学学报, 2020, 40(4):100-108.
WEI S, LIU Y, LIU M N, et al.Purification and identification of PPO from Litopenaeus vannamei and its bioinformatics analysis[J].Journal of Guangdong Ocean University, 2020, 40(4):100-108.
[8] ZHANG Z Y, LI X M, XIAO H, et al.Insight into the allergenicity of shrimp tropomyosin glycated by functional oligosaccharides containing advanced glycation end products[J].Food Chemistry, 2020, 302:125348.
[9] 林娜, 马德蓉, 刘志东, 等.南极磷虾主要过敏原的分离纯化及性质研究[J].水产学报, 2022, 46(3):445-456.
LIN N,MA D R,LIU Z D,et al.Isolation,purification and properties of Antarctic krill(Euphausia superba) major allergens[J]Journal of Fisheries of China,2022,46(3):445-456.
[10] 邵虎明, 肖有明, 谢彦海, 等.鲤鱼小清蛋白多克隆抗体的制备及免疫交叉反应的研究[J].食品科技, 2019, 44(7):50-55.
SHAO H M, XIAO Y M, XIE Y H, et al.Polyclonal antibody preparation and immunological cross-reactivity of carp parvalbumin[J].Food Science and Technology, 2019, 44(7):50-55.
[11] YANG R Q, CHEN Y L, CHEN F, et al.Purification, characterization, and crystal structure of parvalbumins, the major allergens in Mustelus griseus[J].Journal of Agricultural and Food Chemistry, 2018, 66(30):8 150-8 159.
[12] LIU G M, WANG N, CAI Q F, et al.Purification and characterization of parvalbumins from silver carp (Hypophthalmichthy molitrix)[J].Journal of the Science of Food and Agriculture, 2010, 90(6):1 034-1 040.
[13] 朱一丹. 鱼糜发酵过程中致敏蛋白的变化及原因剖析[D].无锡:江南大学, 2020.
ZHU Y D.Changes and causes of allergenic proteins in surimi fermentation[D].Wuxi:Jiangnan University, 2020.
[14] 马涛, 王一侠, 刘艳, 等.超声处理对三文鱼小清蛋白构象及致敏活性的影响[J].食品工业, 2017, 38(3):160-163.
MA T, WANG Y X, LIU Y, et al.Effect of ultrasonic treatment on the antigenicity and conformation of salmon parvalbumin[J].The Food Industry, 2017, 38(3):160-163.
[15] 杨汝晴. 鱼类小清蛋白分离纯化、性质分析及磷酸化改性[D].厦门:集美大学, 2018.
YANG R Q.Isolation, purification, property analysis and phosphorylation modification of fish paralbumin[D]. Xiamen:Jimei University, 2018.
[16] 李弓中, 赵英, 王俊彤, 等.超声处理对蛋清蛋白结构性质及蛋清液起泡性的影响[J].食品科学, 2019, 40(9):68-75.
LI G Z, ZHAO Y, WANG J T, et al.Effect of ultrasonic treatment on protein structure and physicochemical and foaming properties of liquid egg white[J].Food Science, 2019, 40(9):68-75.
[17] RAHAMAN T, VASILJEVIC T, RAMCHANDRAN L.Effect of processing on conformational changes of food proteins related to allergenicity[J].Trends in Food Science and Technology, 2016, 49:24-34.
[18] 陈佳奇, 刘天毅, 贾逾泽, 等.水煮鱼微波烹饪过程中鱼肉水分及组织变化对嫩度的影响[J].食品与机械, 2020, 36(4):51-55.
CHEN J Q,LIU T Y,JIA Y Z,et al.Effects of moisture and tissue changes on tenderness of boiledfish during microwave cooking[J].Food and Machinery,2020,36(4):51-55.
[19] 盛利燚,刘书来,吕飞,等.热煮对丁香鱼肌肉特性及质构的影响[J].食品科学,2011,32(11):5-8.
SHENG L Y, LIU S L, LYU F, et al.Effect of hot boiling on muscle characteristics and textural property of young anchovy (Engraulis japonicus)[J].Food Science, 2011, 32(11):5-8.
[20] MOHSEN M,REZA F,KOBRA M, et al.Identification and characterization of main allergic proteins in cooked wolf herring fish[J].Iranian Journal of Allergy, Asthma, and Immunology, 2016, 15(5):363-371.
[21] 于滢. 几种淡水鱼过敏原小清蛋白的纯化鉴定及性质研究[D].杭州:中国计量学院, 2015.
YU Y.Purification, identification and properties of several freshwater fish allergen paralbumin[D].Hangzhou:China University of Metrology, 2015.
[22] LÓPEZ-FANDIÑO R.Role of dietary lipids in food allergy[J].Critical Reviews in Food Science and Nutrition, 2020, 60(11):1 797-1 814.
[23] 陶淼, 梁提松, 肖朝耿, 等.体外模拟淡水鱼小清蛋白对胃肠消化的影响(英文)[J].农业生物技术学报, 2016, 24(12):1 908-1 914.
TAO M, LIANG T S, XIAO C G, et al.Effect of parvalbumins on gastrointestinal digestion in freshwater fish with in vitro simulation[J].Journal of Agricultural Biotechnology, 2016, 24(12):1 908-1 914.
[24] 张敏. 基于糖基化反应的低致敏鳕鱼小清蛋白制备及其结构研究[D].南昌:南昌大学, 2020.
ZHANG M.Preparation and structure of hypoallergenic cod paralbumin based on glycosylation reaction[D].Nanchang:Nanchang University, 2020.
[25] DUPONT D, MANDALARI G, MOLLE D, et al.Comparative resistance of food proteins to adult and infant in vitro digestion models[J].Molecular Nutrition Food Research, 2010, 54(6): 767-780.
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