【摘要】 以牛血球为材料,经溶血等处理和丙酮沉淀,获得牛血球超氧化物歧化酶粗品。此粗酶可以通过DEAE-Sepharose和CM-Sepharose快速柱层析,获得超氧化物歧化酶纯品。纯化的酶比活可达13500μ/mg,经PAGE、SDS-PAGE和快速蛋白液相色谱(FPLC)检测,结果表明,纯化酶是均一的。Sephadex G-100凝胶过滤测得该酶分子量为31,800,SDS-PAGE测得亚基分子量为15,800;N-末端氨基酸是丙氨酸;氨基酸组成分析及色氨酸含量测定表明,该酶由290个氨基酸组成,不含色氨酸,紫外吸收光谱的最大吸收峰在257nm,对酶稳定性的研究表明,该酶是一种稳定性很高的酶,对温度、pH和某些变性剂表现出很高的耐受性。更多还原

【Abstract】 Superoxide dismutase from bovine erythrocyte can be purified by a procedure of hemolysate, ethanol-chloroform treatment, K2HPO4 and acetone precipitate, DEAE-Sepharose (Fast Flow)chromatograph and CM-Sepharose chromatograph. The purified enzyme is found to be homogeneous by the analysis of SDS-PAGE, PAGE and FPLC. The specific activity of purified enzyme is 13500μ/mg,molecular weight is 31,800 by gel filtration,subunit weight is 15,800 by SDS-PAGE. The N-terminal amino acid is determined to be Ala by DNS method. Analysis of amino acid composition of SOD shows that it is consist of 290 amino acid residues and no Trp. The maximum UV peak of SOD is at 257nm. The enzyme is relatively stable in acid,alkali,heat treatment and some denaturants.更多还原