摘要
目的从羊红细胞中分离纯化铜锌超氧化物歧化酶 (Cu ,Zn SOD) ,并对其部分理化性质进行研究。方法采用有机溶剂去除血红蛋白、热变性、超滤浓缩、丙酮沉淀、DE FF柱色谱的方法 ,对羊红细胞Cu ,Zn SOD进行分离纯化。结果羊红细胞 5 2 0g得Cu ,Zn SOD总活力为 4 6 70 0 0u ,比活为 812 6u/mg·pro,纯化倍数为 2 6 .2 ,活性回收率为 6 1%。理化性质分析表明 :该酶的最大紫外吸收波长为 2 5 8nm ,亚基相对分子量为 16 .1kD ,每个亚基含 1个铜原子和 1个锌原子。pH在 6～ 11范围内该酶稳定性很好 ,在 35℃～ 75℃范围内 ,保温 2 0min ,酶活基本没有损失。 2mmol/L的SDS对Cu ,Zn SOD没有明显的抑制作用 ,而 2mmol/L的H2 O2 对该酶表现出较强的抑制作用。结论采用此方法分离纯化的Cu ,Zn SOD达到电泳纯 ,其理化性质与其它动物血液来源的Cu ,Zn SOD一致。
PurposeTo purify Cu,Zn SOD from sheep erythrocyte and to study some properties of purified Cu,Zn SOD.MethodsCu,Zn SOD was purified from sheep erythrocyte by organic solvent to remove hemoglobin, thermal denaturalization, super filtration condensation, acetone precipitation and DE FF chromatography.ResultsThe purified enzyme obtained from 520 gram of sheep erythrocyte possessed a total activity of 467 000 units, a specific activity of 8 126 u/mg protein was reached,corresponding to 26.2 fold purification. The yield was 61%. The investigation of physiochemical properties of the SOD showed that the ultraviolet absorption wavelength of the enzyme was found to be at 258 nm.The molecular weight of the subunit was 16.1 kD. The purified SOD contained 1.0 atom of Cu and 1.0 atom of Zn per subunit. The enzyme was found to have good stability at the pH range 6～11 and displayed good thermal stability at the 35℃～75℃ range over 20 minutes incubation period. The SOD enzyme was not obviously inhibited by 2mmol/L SDS, but obviously inhibited by 2mmol/L H 2O 2.ConclusionThe purity of Cu,Zn SOD obtained by this process was electrophoretically pure. Cu,Zn SOD from sheep erythrocyte had the similar properties to other Cu,Zn SOD from other animal blood.
出处
《中国生化药物杂志》
CAS
CSCD
2003年第1期4-7,共4页
Chinese Journal of Biochemical Pharmaceutics
关键词
羊红细胞
铜锌超氧化物歧化酶
纯化
性质
sheep erythrocyte
copper and zinc superoxide dismutase
purification
properties
