苏云金杆菌4.0718菌株超氧化物歧化酶的纯化和性质研究

苏云金杆菌4.0718菌株超氧化物歧化酶的纯化和性质研究 Studies on Purification and Characterization of Superoxide Dismutase from Bacillus thuringiensis Strain 4.0718 李伟 LI Wei 1 1 2 3 4 5 first-author 夏立秋 3 1 2 3 4 5 周源 XIA Li-qiu 2 1 2 3 4 5 王先酉 ZHOU Yuan 3 1 2 3 4 5 邹国林 WANG Xian-you 3 1 2 3 4 5 武汉大学生命科学学院 武汉大学生命科学学院 College of Life Sciences, Wuhan University College of Life Sciences, Wuhan University 湖南师范大学生命科学学院 湖南师范大学生命科学学院 College of Life Science, Hunan Normal University College of Life Science, Hunan Normal University 湖南师范大学医学院 湖南师范大学医学院 College of Medicine, Hunan Normal University College of Medicine, Hunan Normal University 武汉大学生命科学学院,湖北,武汉,430072 武汉大学生命科学学院,湖北,武汉,430072 ,湖南,长沙,410082 ,湖南,长沙,410082 经 （NH4 ） 2 SO4 分级沉淀、SephadexG 75凝胶过滤和DEAE 5 2柱层析将苏云金杆菌 4.0 718菌株的超氧化物岐化酶 （SOD）纯化到均一程度 ,酶比活力达 688.0U/mg ,酶得率为 47.9% 该酶经KCN ,H2 O2 ,氯仿 乙醇抑制实验 ,金属元素含量测定和紫外可见吸收光谱测定 ,表明是Fe SOD 经反相高效液相色谱和电喷雾质谱联合分析等方法 ,测得酶分子量为 3 9.4861× 10 6 ,由两个相同亚基组成 ,N 末端氨基酸为丙氨酸 A superoxide dismutase from Bacillus thuringiensis strain 4.0718 has been purified to homogeneity by ammonium sulfate fractionation, Sephadex G-75 gel filtration and DEAE-cellulose-exchange chromatograph. The specific activity of the purified enzyme is 688.0 U/mg protein and the yield of the purified enzyme is 47.9%. The enzyme is an iron superoxide dismutase as assayed by its sensitivity to inhibitor: KCN, H 2O 2, CH 3CH 2OH-CHCl 3 and other methods: metal elements measure and ultraviolet-visible spectroscopy. The enzyme has a molecular weight of 39.486 1×10 6 with conjoint analysis of reversed-phase high-performance liquid chromatography and electrospray ionzation mass spectrometry. The enzyme contains two equally sized subunits.The N-terminal amino acid of the enzyme is alanine as tested by DNS-Cl. 超氧化物歧化酶 苏云金杆菌 纯化 性质 superoxide dismutase Bacillus thuringiensis purification characterization Q554 国家自然科学基金资助项目 （ 30 2 70 0 37） 2003-02-01 2021-04-01 2