重组Hepcidin融合蛋白的氧化复性及活性鉴定  

Oxidative Refolding and Biological Activity Detemination of Recombinant Hepcidin Fusion Protein

作　　者：王继雯[1,2] 谢宝恩[1,2] 甄静[1] 周伏忠[1] 陈国参[2] 

机构地区：[1]河南省微生物工程重点实验室,郑州450008 [2]河南省科学院生物研究所,郑州450008

出　　处：《生物技术通报》2010年第5期192-195,共4页Biotechnology Bulletin

基　　金：河南省高新领域重点攻关计划项目(082102220002)

摘　　要：将诱导表达的His-hepcidin融合蛋白包涵体通过固定化金属离子配体亲和层析(IMAC)柱分离纯化后,在cys-teine/cystine氧化还原体系中氧化形成二硫键,稀释复性后用肠激酶将融合蛋白的his-tag切除。酶切后所得的Hepcidin经抑菌圈试验检验,对枯草芽孢杆菌等革兰氏阳性菌和部分革兰氏阴性菌具有抗菌活性。After isolation and purification with the IMAC column, the His-hepcidin fusion protein was oxidized and renaturated in cysteine/cystine system to form disulfide bond. The his-tag was excised by enterokinase and the purity Hepcidin protein was up to 90%. Through agar diffusion assay, the recombinant hepciitin displayed obvious antibacterial activity against Gram bacteria such as B. subtilis and some negative bacteria.

关 键 词：HEPCIDIN 纯化 氧化复性 抑菌圈 

分 类 号：Q78[生物学—分子生物学]