摘要
用乙醇—氯仿处理、丙酮分级沉淀、ＤＥＡＥ—纤维层析等方法，从兔血红细胞中纯化出钢锌超氧化物歧化酶（ＳＯＤ），产率为５１％，比活力为１２４３０ｕ／ｍｇ·该酶经冷冻干燥后呈淡蓝色．天然状态下，该酶的分子量约为３００００，由相同的两个亚基组成．紫外扫描表明纯酶在２６５ｎｍ处有最大光吸收值．经聚丙烯酸胺凝胶电泳后，用蛋白染色及ＮＢＴ活性染色均显示出三条带．结果表明用该纯化方法得到的ＳＯＤ为均一性纯酶．氨基酸组成分析及元素分析表明兔血铜锌与采用其他来源的酶基本相同．化学修饰结果表明兔血铜锌ＳＯＤ没有游离巯基．
Superoxide dismutase (SOD) was purified from rabbit red cells by with McCord and Fridovich method. The Cu-Zn SOD is of a specific acitivity of 12000u/mg. The percentage yield is ca. 51%. The CuZn-SOD is of a pale blue -green color. In its natural form the molecular weight is ca. 30 kDa, consisting of two subunit. The enzyme exhibits an ultraviolet spectrum with a maximum at 265 urn. NBT activity staining and Coomassie Blue staining show the same three bands of the purified enzyme by PAGE. The results show that the CuZn-SOD purified by this method is homogenously pure. The amino acid anlyysis and atom absorption spectrum of the enzyme is very similar to CuZn-SODs reported from other sources. There is no free sulfhydryl group in the enzme.
出处
《复旦学报（自然科学版）》
CAS
CSCD
北大核心
1994年第5期577-583,共7页
Journal of Hehai University：Natural Science
关键词
血液
铜
锌
超氧化物歧化酶
兔血
纯化
CuZn superoxide dismutase
rabbit erythrocyte
purification
