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通过对甘蓝细胞溶质SOD的纯化及性质研究 ,开拓制备SOD的廉价原料来源。经硫酸铵沉淀、SephadexG 10 0凝胶过滤和DEAE DE5 2层析 3个步骤 ,将甘蓝细胞溶质SOD纯化到均一程度。从 10 0 0 g叶片中得到 3 2mg酶 ,酶的比活力达 970 0U/mg蛋白。经鉴定该酶是Cu·Zn SOD。测得其Mr 约为 32 2 0 0 ,亚基Mr 约为 160 0 0。该酶的N 末端为丙氨酸 ,其可见光区与紫外光区的最大吸收峰分别在 680nm和 2 65nm。氨基酸组成分析表明甘蓝细胞溶液质SOD由2 66个氨基酸残基组成 ,而且不含色氨酸。本研究对高等植物细胞溶质SOD的纯化建立了一种行之有效的方法 ,为高等植物SOD性质的比较研究提供了实验依据。
Through the purification and characterization of SOD in cabbage cytosol, the source of cheap raw materials for preparing SOD was explored. The cell lysates were purified to homogeneity by ammonium sulfate precipitation, Sephadex G 10 0 gel filtration and DEAE DE 5 2 chromatography. 32 mg of enzyme was obtained from 10 0 g leaves with a specific activity of 970 0 U / mg protein. The enzyme was identified as Cu · Zn SOD. Mr measured about 3220, Mr subunit about 160 0 0. The N-terminal of the enzyme is alanine, and the maximum absorption peak in the visible light region and ultraviolet light region is 680 nm and 265 nm, respectively. Amino acid composition analysis showed that SOD in cabbage cell solution consisted of 266 amino acid residues, and did not contain tryptophan. This study established an effective method for the purification of higher plant cytosolic SOD and provided experimental evidence for the comparative study of the SOD properties of higher plants.