组氨酸标签蛋白纯化介质——EDTA-壳聚糖的制备与应用  

The Purification of Histidine Tag Protein Medium-Preparation of EDTA-Chitosan and Its Application

作　　者：邵明聪 华伟伟[1] 邱金丹 黄建颖[1] Shao Mingcong;Hua Weiwei;Qiu Jindan;Huang Jianying(School of Food Science and Biotechnology,Zhejiang Gongshang University,Hangzhou 310018)

机构地区：[1]浙江工商大学食品与生物工程学院,杭州310018

出　　处：《中国食品学报》2018年第12期144-149,共6页Journal of Chinese Institute Of Food Science and Technology

基　　金：浙江省自然科学基金项目(Y3110204);浙江省重大科技专项(2011C12031);国家自然科学基金项目(21102129)

摘　　要：以离子液体为溶剂,溶解具有较强金属螯合能力的EDTA-壳聚糖,能够保持较强的稳定性。将其用于涂膜处理,螯合两种金属离子(Ni2+,Zn2+),形成M2+-EDTA-壳聚糖(M=Ni,Zn),并对组氨酸标签蛋白进行特异性亲和纯化。结果表明:EDTA-壳聚糖经过涂膜处理后,形成坚韧而富有弹性的膜,用于组氨酸蛋白分离取得良好的效果,相比直接用于EGFP-(His)6-NH2蛋白纯化分离表现很大的优势,得到目的蛋白的纯度约为95%,与涂膜前相比,缩短了10 min的分离时间,且能更好地特异性吸附目的蛋白,在一定程度上节约了时间和成本。Using ionic liquids as the extraction solvents to solve EDTA-Chitosan,which has good ability of metal chelation,it enhances the products'stability.When we take operation of coating film and chelating different metal ions (Ni^2+,Zn^2+),finally it forms the M^2+-EDTA-Chitosan (M=Ni,Zn).And then,M^2+-EDTA-Chitosan (M=Ni,Zn)is used to purify histidine tag protein relying on the affinity resulted from the complexes chelated by different metal ions (Ni^2+,Zn^2+). The results shows that EDTA-Chitosan has formed a steel and elastic film by coating.Furthermore,the process of purifying the His-tagged protein achieved a good effect.Compared with the directly purification of the EGFP-(His)6-NH2,it shows a large advantage,we eventually obtained the purity of about 95% of the objective protein.And compared to the prior of coating,it shorts the separation time about 10 min,absorbs the objective protein .accurately,and saves the time and expense certainly.

关 键 词：离子液体 EDTA-壳聚糖 组氨酸标签 分离纯化 

分 类 号：Q503[生物学—生物化学]