以聚乙二醇为层析伴侣同时制备SOD、过氧化氢酶和血红蛋白  

Simultaneous Purification of Superoxide Dismutase, Catalase and Hemoglobin from Bovine Erythrocyte Lysate with PEG 600 as Chaperon

作　　者：王永权[1] 路秀玲[1] 苏志国[1] 

机构地区：[1]中国科学院过程工程研究所生化工程国家重点实验室,北京100080

出　　处：《生物工程学报》2005年第3期466-472,共7页Chinese Journal of Biotechnology

基　　金：国家自然科学基金重点基金项目 (No.2 0 13 60 2 0 );中国科学院知识创新重点方向项目~~

摘　　要：发展了一条从红细胞裂解液中同时制备超氧化物歧化酶(SOD)、过氧化氢酶和血红蛋白的新工艺。采用0 75 %的聚乙二醇6 0 0作为层析伴侣,使血红蛋白直接流过阴离子交换层析柱,同时吸附SOD和过氧化氢酶。经过梯度洗脱获得SOD和过氧化氢酶组分,再经过疏水性相互作用层析与凝胶过滤层析相串联,使SOD和过氧化氢酶得到纯化。纯化后的SOD和过氧化氢酶的比活力分别达到15 932u mg和6 5 918u mg ,血红蛋白的纯度达到99 9%以上。总回收率为:SOD ,4 7 4 % ;过氧化氢酶,2 9 6 % ;血红蛋白,88 7%。Superoxide dismutase, catalase and hemoglobin were purified simultaneously from the same batch of bovine erythrocyte lysate. The process involves an initial anion exchange chromatography, followed by a hydrophobic interaction chromatography and gel filtration chromatography. 0.75% polyethylene glycol 600 was added as a purification chaperon before the anion exchange chromatography. The hemoglobin fraction passed through the ion exchange column without being retained. The superoxide dismutase and catalase were adsorbed by the column and were eluted separately during elution. The two eluted fractions containing crude superoxide dismutase and catalase were further purified with hydrophobic interaction chromatography and gel filtration chromatography in sequence. The specific activities of superoxide dismutase and catalase were 15932u/mg and 65918u/mg, respectively. SDS-polyacrylamide gel electrophoresis and gel filtration chromatography were used to analyze the purity of the proteins. The purity of superoxide dismutase, catalase and hemoglobin were 77.6%, 81.9% and 99.9%, respectively. The total recoveries for superoxide dismutase, catalase and hemoglobin were 47.4%, 29.6% and 88.7%, respectively.

关 键 词：超氧化物歧化酶 过氧化氢酶 血红蛋白 离子交换层析 疏水性相互作用层析 

分 类 号：TQ463[化学工程—制药化工]