毕赤酵母表达重组人源Cu/Zn SOD的分离纯化与稳定性表征  

Purification of Recombinant Human Cu/Zn SOD Expressed in Pichia Yeast and Its Stability Characterization

作　　者：杨志强[1] 张晨[1] 敖伟[1] 蔡泽恩[1] 刘树滔[1] 饶平凡[1] 

机构地区：[1]福州大学生物工程研究所,福州350002

出　　处：《中国食品学报》2011年第5期108-113,共6页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金项目(No.31071497);福建省卫教联合攻关项目(WKJ2008-2-47);福建省高等学校新世纪优秀人才支持计划资助项目(SW2006-15)

摘　　要：通过乙醇沉降、强阳离子交换、强阴离子交换等纯化方法对毕赤酵母表达的重组人源Cu/ZnSOD进行分离,并测定该纯化蛋白的热稳定性和pH稳定性。试验结果表明通过这些方法获得比活为12881U/mg的目标蛋白,该蛋白达到反相色谱纯。纯化后的SOD蛋白无论在低于75℃或在pH3～10条件下放置2h,均能保持85%以上的活力。采用毕赤酵母体系生产SOD,产量大,纯化成本低,蛋白稳定性高,有利于重组人源Cu/ZnSOD的大规模生产及应用。Conventional methods for purification,including alcohol sedimentation,Strong cation exchange and strong anion exchange,were carried out to purify SOD,which were expressed by Pichia yeast system.Then the sensitive to temperature and pH condition of purified SOD were determined.Data showed that large quantity of SOD proteins,which were detected for only one peak by reversed phase chromatography,can be obtained by these methods.The activity of these proteins can reserved more than 85%,while they were incubated in the condition of under 75 ℃ or during pH 3～10.It is obviously that Pichia yeast system can produce large quantities of SOD proteins with low cost and increase the stability of SOD,which are in favour of industrial manufacture and SOD applications.

关 键 词：毕赤酵母 Cu/ZnSOD 纯化 稳定性 

分 类 号：TQ464.7[化学工程—制药化工]