啤酒酵母生产的重组水蛭素的纯化及脱色  

Purification and Decolorization of Recombinant Hirudin Produced by Saccharomyces Cerevisiae

作　　者：昝云红[1] 孙茂盛[1] 李智华[1] 徐彤浩[1] 李健峰[1] 姬秋彦[1] 马雁冰[1] 戴长柏[1] 

机构地区：[1]中国医学科学院中国协和医科大学医学生物学研究所,昆明650107

出　　处：《中国生物化学与分子生物学报》1999年第5期779-781,共3页Chinese Journal of Biochemistry and Molecular Biology

基　　金：云南省应用基础研究基金!(青年基金项目 ) ( 98C0 2 3 Q)

摘　　要：对啤酒酵母生产的重组水蛭素变异体1(rHV1)进行多步骤的纯化。首先将分泌到培养上清液中的水蛭素进行硫酸铵沉淀和SephadexG-50凝胶过滤,再用Q-SepharoseHP阴离子交换层析分离,最后用HPLCSP-5PW阳离子交换柱脱色及HPLCC8柱反相层析。真空干燥后得到的水蛭素蛋白经SPS-PAGE、N端氨基酸序列分析、抗凝血酶活力分析鉴定,证明已获得高纯度的重组水蛭素HV1制剂。Recombinant hirudin produced by Saccharomyces cerevisiae was highly purified Hirudin expressed by yeast and secreted to culture supernatant was first precipitated by(NH 4) 2SO 4 and filtrated by Sephadex G 50 Then it was isolated by Q Sepharose HP anion exchange chromatography Last it was decolourized by HPLC cation exchange chromatography of SP 5PW column After reverse phase HPLC of C8 column and vacuum drying,the purely white hirudin protein was obtained.It was proved to be correct recombinant hirudin HV1 by SDS PAGE,amino acid sequence analysis of N terminal and activity identification of antithrombin The results supplied evidence for scale production and clinical application of recombinant hirudin produced by gene engineering method

关 键 词：啤酒酵母 重组水蛭素 纯化 脱色 

分 类 号：Q784[生物学—分子生物学] Q513