柞蚕蛹超氧化物歧化酶的提取及性质的研究初探

PRELIMINARY REPORT OF EXTRACTION TECHNIQUE OF SUPEROXID DISMUTASE FROM THE PUPAE OF AUTHERAEA PERNYI

从柞蚕蛹中分离铜锌超氧化物歧化酶(Cu,Zn-SOD),并对其部分性质进行研究。用氯仿-乙醇除杂蛋白质、丙酮沉淀、热变性、DE-32柱色谱等方法,对柞蚕蛹Cu,Zn-SOD进行分离纯化。结果得Cu,Zn-SOD干粉9.5mg;比活10116.7U/mgpro.;活力回收55.9%;纯化倍数为360.7。该酶最大吸收波长258nm;pH在5～9范围内基本稳定;在40℃～60℃内保温30min酶活基本不变;H2O2对此酶有明显的抑制作用。柞蚕蛹含有丰富的Cu,Zn-SOD。

To purify cupper / zinc superoxide dismutase （Cu,Zn-SOD） from Autheraea pernyi＇s pupae to study some properties of purified Cu,Zn-SOD. Cu,Zn-SOD has been purified from Autheraea pernyi＇ s pupae by chloroform-ethanol fractionation ,acetone precipitation, thermal denaturalization and DE-32 chromatography. 9.5 mg of dry powder enzyme was obtained ,the purified specific activity of the enzyme was 10 116.7 U/mg protein, the yield was 55.9 % , and the ultraviolet absorption peak of theenzyme was found at 258 nm. Its purification factor was 360.7.This enzyme has two identical subunits.The enzyme was found to have good stability at the pH range 5 to 9 and displayed good thermal stability at the 40 ℃-60 ℃ range over 30 min incubation period. The Cu,Zn-SOD was obviously inhibited by H2O2.