摘要
本文报道了制备超氧化物歧化酶的方法,即首先75℃下加热处理15min,然后用硫酸铵分级盐析,再用少量乙醇、氯仿处理,最后经分子筛层析,可得到高纯度的产品。从1000ml猪红细胞中,可得到46.5mg酶,比活力达每毫克蛋白14200u.1000ml码红细胞,可得到30mg酶,比活力达每毫克蛋白16600u.所得产品经disc-PAGE,SDS-PAGE,N-末端氨基酸分析等检测,均表明已达均一程度。此法可减轻环境污染,操作简便,经济实用,适用于工厂化生产。
Copper/Zinc superoxide dismutase from hog or chicken erythrocytes was isolated and purified by a simple and cheap method: 75℃ heating for 15min,(NH_4)_2SO_4 salting out, organic reagent reacting and Sephadex G-100 chromatographing. The purified enzymes were found to be homogeneous as judged by disc-PAGE, SDS-PAGE and the analysis of N-terminal amino acid. The purified SOD obtained from one liter of porcine red cells contained a total of 46.5mg of enzyme with a specific activity of 14200 units permg of protein. The purified SOD obtained from one liter of chicken red cells contained a total of 30mg of enzyme with a specific activity of 16600 units per mg of protein.
关键词
超氧歧化酶
血红细胞
制备
Superoxide dismutase
Erythrocyte
New preparation method
