重组人锰超氧化物歧化酶纯化条件研究  

Study on Purification Condition of Recombinant Human Manganese Superoxide Dismutase

作　　者：程贻[1] 陈车生[2] 吴乾一[2] 赵健[1] 袁勤生[1] 

机构地区：[1]华东理工大学生物反应器国家重点实验室,上海200237 [2]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《食品与药品》2007年第07A期1-5,共5页Food and Drug

摘　　要：目的研究重组人锰超氧化物歧化酶(rhMn-SOD)的纯化条件。方法超声法破碎细胞,用热变性方法沉淀蛋白质;优化DEAE离子交换层析的的分离条件,并用葡聚糖凝胶G100分子筛分离纯化蛋白质。结果菌体破碎最佳超声功率为300W,超声70个循环,粗酶液热处理最佳温度为75℃,时间10 min;DEAE上样缓冲液最佳pH值为9.0,洗脱液NaCl含量为0.1 mol/L,G100纯化后,rhMn-SOD比活达到4 411.706 U/mg,纯化倍数是粗酶液的5倍。结论得到了适于分离纯化rhMn-SOD的较简便的工艺。Objective To study the purification method of recombinant human manganese superoxide dismutase （rhMn-SOD）. Methods The cells were fragmented by ultrasonic. The protein was precipitated by the thermal denaturation. The condition of proteins separation by DEAE ion exchange chromatography was optimized, then Sephadex G-100 molecular sieving was used to purify protein. Results The optimization condition of ultrasonic power was 300 W, the cycles of ultrasonic were 70 cycles, the best thermal denaturation was 75 ℃ and time was 10 minutes. The optimal pH value of loading buffer solution of DEAE was 9.0, the elution buffer was 0.1mol/ LNaCl. After G-100, the specific activity of rhMn-SOD wa℃s 4 411.706 U/mg and it was 5-fold higher than crude enzyme extract. Conclusion The convenient method of separation and purification of rhMn-SOD is obtained.

关 键 词：重组人锰超氧化物歧化酶 大肠杆菌 离子交换层析 

分 类 号：Q554[生物学—生物化学]