组氨酸标签蛋白纯化介质的合成及应用

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英文篇名：Synthesis of Purification Media of the Histidine Tag Protein and Its Application 作者：华伟伟 ; 徐维元 ; 黄建颖 ; 杨利军 英文作者：Hua Weiwei;Xu Weiyuan;Huang Jianying;Yang Lijun;School of Food Science and Biotechnology, Zhejiang Gongshang University, Food Processing and Preservation Key Lab; 关键词：亲和性离子液体 ; 螯合 ; 组氨酸标签 ; 蛋白质纯化 英文关键词：affinity ionic liquid;;chelate;;histidine tag protein;;purification of protein 中文刊名：ZGSP 英文刊名：Journal of Chinese Institute of Food Science and Technology 机构：浙江工商大学食品与生物工程学院浙江省果蔬保鲜与加工技术研究重点实验室; 出版日期：2016-12-06 16:19 出版单位：中国食品学报 年：2016 期：v.16 基金：浙江省重大科技专项(2011C12031);; 国家自然科学青年基金项目(21102129) 语种：中文; 页：ZGSP201610011 页数：7 CN：10 ISSN：11-4528/TS 分类号：85-91

摘要

以离子液体为萃取溶剂,将氮杂大环功能性配体与疏水性离子液体链接构建亲和性离子液体,通过氮杂大环螯合金属离子构成复合物;利用螯合的金属离子(Cu2+、Ni2+、Zn2+)对组氨酸标签特异的亲和性,对组氨酸标签蛋白进行特异性亲和纯化,结果表明:AIL螯合金属离子后才会对FITC-(His)6-OH产生亲和性。Cu-AIL亲和性最强,对于His-tag EGFP的选择性最差;Ni-AIL受限于其低的金属离子结合量,获得的目的蛋白最少;Zn-AIL能有效亲和His-tag EGFP,纯化效率相对最佳。结合以上不同亲和介质的特性,开发出利用Zn-AIL与Cu-AIL的二次纯化体系,获得纯度约90%的His-tag EGFP。
        Synthesize affinity ionic liquid(AIL) through linking polyazamacrolyle with hydrophobic ionic liquid based on the designability using ionic liquids as extraction solvents. Affinity ionic liquid(AIL) is used for the purification of histidine tag protein relying on the affinity resulted from the complexes chelated by metal ion(Cu2 +、Ni2 +、Zn2 +) and polyazamacrocyle. The results showed that only when AIL chelated by metal ion, can it attach to the FITC-(His)6-OH.Cu-AIL exhibited the best affinity ability for his-tag EGFP but selective ability was the worst one; Ni-AIL was limited by its low amount of metal ion combination, so the purpose protein is the least; Zin-AIL could attach the his-tag EGFP efficiently, possessing the best purification efficiency. And combined with the above different affinity media features, we developed the secondary purification system with Zn-AIL and Cu-AIL, obtained the purity of about 90% of His-tag EGFP.

引文

[1]王子佳,李红梅,弓爱君,等.蛋白质分离纯化方法研究进展[J].化学与生物工程,2009,26(8):8-11.
    [2]牛瑞,秦胜利.蛋白质分离纯化技术研究进展[J].化工科技市场,2010,33(4):16-18.
    [3]王红颖,王倩,黄瑾.蛋白质分离纯化方法的研究进展[J].现代生物医学进展,2011(s2):5168-5170.
    [4]刘卉,陶国宏,邵元华,等.功能化的离子液体在电化学中的应用[J].化学通报,2004,67(11):795-801.
    [5]杜平,胡维,李胜清,等.离子液体在萃取分离中的应用研究进展[J].分析科学学报,2009,25(5):598-604.
    [6]李琼.固定化金属离子螯合琼脂糖凝胶的制备及其纯化组氨酸标签蛋白的研究[D].南京:南京理工大学,2012.
    [7]Hochuli E,Bannwarth W,Dbeli H,et al.Genetic approach to facilitate purification of recombinant proteins with a novel metal chelate adsorbent[J].Nature Biotechnology,1988,6(11):1321-1325.
    [8]Jain P,Baker GL,Bruening ML.Applications of polymer brushes in protein analysis and purification[J].Annual Review of Analytical Chemistry,2009,2(2):387-408.
    [9]Jiang W,Graham B,Spiccia L,et al.Protein selectivity with immobilized metal ion-tacn sorbents:chromatographic studies with human serum proteins and several other globular proteins[J].Analytical Biochemistry,1998,255(1):47-58.
    [10]Tseng MC,Tseng MJ,Chu YH.Affinity ionic liquid[J].Chemical Communications,2009,48:7503-7505.
    [11]Graham B,Spiccia L,Skelton BW,et al.Imidazole derivatives of binuclear copper(II)and nickel(II)complexes incorporating bis(1,4,7-triazacyclononan-1-yl)ligands[J].Inorganica Chimica Acta,2005,358(13):3974-3982.
    [12]Kytepe S,Erdoggan S,Sekin T.Synthesis of polyimide from 5,5''-bis(bromomethyl)-2,2',6',2''-terpyridine and investigation of the polymer sorption behavior towards some metal ions[J].Journal of Hazardous Materials,2009,162(2):695-702.
    [13]You L,Wu Z,Kim T,et al.Kinetics and thermodynamics of bromophenol blue adsorption by a mesoporous hybrid gel derived from tetraethoxysilane and bis(trimethoxysilyl)hexane[J].Journal of Colloid and Interface Science,2006,300(2):526-535
    [14]邹泽红.BCA法测定蜜蜂变应原中的蛋白含量[J].化学分析计量,2004,13(5):52-53.
    [15]Rahnemaie R,Hiemstra T,van Riemsdijk WH.A new surface structural approach to ion adsorption:tracing the location of electrolyte ions[J].Journal of Colloid and Interface Science,2006,293(2):312-321.