黑脉羊肚菌SOD的纯化及特性研究

Purification and characteristics of SOD in Morchella angusticeps

采用热击法和硫酸铵分级沉淀法对所提取的黑脉羊肚菌SOD进行纯化，对纯化后酶的热稳定性、pH稳定性、敏感性及同工酶类型等进行研究。结果表明：热击法最佳热击温度为60℃，最佳热击时间为15min；硫酸铵分级沉淀法最佳盐溶条件是采用40％饱和度硫酸铵，最佳盐析条件是采用85％饱和度硫酸铵；所纯化的SOD表现出良好的热稳定性和pH稳定性，在60℃下保存60min，其酶活保留超过50％，pH适应范围为6～9；Mn2＋对羊肚菌SOD有明显的激活作用，Fe2＋对其有明显的抑制作用；该SOD能耐受过氧化氢，但对氟仿-乙醇和SDS较为敏感，黑脉羊肚菌SOD属于Mn-SOD类型。

SOD in Morchella angusticeps was purified by heating and ammonium sulfate graded precipitation.Then, the thermal stability,pH stability,sensitivity and isozyme types of SOD were also studied.Results showed that the best temperature and time of heating method was 60 ℃ and 15 min, respectively.The optimum mass fraction of ammonium sulfate for salt-soluble and salting-out were 40% and 85%, respectively.The purified SOD showed good thermal stability and pH stability. SOD retained more than 50% when save 60 min at 60 ℃ and the accommodation pH was 6-9.Mn2＋ could significantly activate purify SOD,while Fe2＋ remarkably inhibit the activity of SOD.The SOD could tolerate hydrogen peroxide, however, it was sensitive to chloroform-ethanol and SDS. Therefore,this SOD belongs to Mn-SOD type.