Q Sepharose^(TM) XL强阴离子交换色谱快速纯化猪心中肌红蛋白和细胞色素C  

Fast Purification of Myoglobin and Cytochrome-C from Pig Heart via Q Sepharose^(TM) XL Strong Anion Exchange Chromatography

作　　者：李琳子[1] 徐朋朋[1] 李任强[1] 

机构地区：[1]暨南大学生物工程学系,广东广州510632

出　　处：《化学与生物工程》2009年第10期56-59,共4页Chemistry & Bioengineering

基　　金：暨南大学自然科学基金资助项目(640073)

摘　　要：以Q SepharoseTM XL强阴离子交换色谱结合Sephadex G-75分子排阻色谱快速提取纯化猪心中肌红蛋白(Myoglobin,Mb)和细胞色素C(Cyt C)。在pH值为7.6~8.5范围内,Cyt C吸附在Q SepharoseTMXL强阴离子交换柱上,而Mb会缓慢通过。所以选择QSepharoseTMXL强阴离子交换柱的上样pH值为7.9,洗脱pH值为7.2。电泳检测证明,经过QSepharoseTMXL柱初步纯化所得Mb和Cyt C的产率分别为89.7%和93.2%;再经过Sephadex G-75分子排阻色谱可得到纯的Mb和Cyt C,其产量分别为77.43mg.(kg猪心)-1和86.42mg.(kg猪心)-1。使用QSepha-roseTMXL强阴离子交换色谱能从猪心中快速制备纯的Mb和Cyt C。A strong anion exchange chromatography Q SepharoseTM XL coupled with Sephadex G-75 sizeexclusion chromatography were adopted to isolate myoglobin （Mb） and cytochrome-C （Cyt C） from pig heart. In determination of the pH value working range of Q Sepharose^TM XL column, Cyt C was found to be captured on Q Sepharose^TM XL column while Mb passed slowly through it at the pH value from 7.6 to 8.5. Thus the protein crude extracts were loaded into Q Sepharose^TM XL column at pH value of 7.9 and the target proteins were eluted at pH value of 7.2. As determined by SDS-PAGF4, the yields of Mb and Cyt C in the extracts, which were obtained in initial purification process by Q Sepharose^TM XL column, were 89.7 % and 93.2% , respectively. The＇pure Mb and Cyt C were then obtained via Sephadex G-75 size-exclusion chromatography. The final productivities of Mb and Cyt C were 77.43 mg·kg^- 1 and 86.42 mg· kg^- 1 , respectively. This study had demonstrated it was available for fast preparation of Mb and Cyt C from pig heart by Q Sepharose^TM XL strong ainon exchange chromatography.

关 键 词：蛋白纯化 Q SepharoseTM XL强阴离子交换色谱 肌红蛋白 细胞色素C 猪心 

分 类 号：TQ464.8[化学工程—制药化工] O657.7[理学—分析化学]