云芝菌丝体超氧化物歧化酶的分离纯化及性质研究

Study on purification and characterization of superoxide dismutase in Trametes versicolor mycelium

摘要: 从云芝菌丝体中分离纯化超氧化物歧化酶（SOD）,并对其性质进行了研究。结果表明,SOD粗酶液经膜过滤、硫酸铵分级盐析、SephadexG-75凝胶柱层析分离后,SOD纯化了12.6倍,总回收率为50.6%,获得SOD酶比活为4.61U/mg蛋白。提纯SOD在257nm处有特征吸收峰,其活性受氯仿-乙醇影响不明显,但受到H2O2明显抑制,酶活在50℃以下较稳定,超过50℃以上,活性随温度升高而降低。说明云芝菌丝体SOD以Cu,Zn离子为辅基,并具有较好的热稳定性。

Abstract: Superoxide dismutase ( SOD) was extracted from Trametes versicolor mycelium with membrane filtration, ammonium sulfate fractionation and SephadexG-75 gel column chromatography. The SOD was purified 12.6-fold with a specific activity of 4.61U/mg protein and a yield of 50.6%. The ultraviolet spectrum of purified SOD showed an absorbance peak at 257nm. The activity of enzyme was stable under 50℃ but decreased with higher temperature. Meanwhile, it was sensitive to chloroform-enthanol but inhibited obviously by H2 O2. These results indicated that the SOD from Trametes versicolor mycelium was Cu, Zn-SOD with a good thermostability.