离子交换法纯化罗非鱼血超氧化物歧化酶的研究

Research on purification conditions of superoxide dismutase from tilapia blood by ion exchange chromatography

为使罗非鱼血超氧化物歧化酶(SOD)的纯化工艺条件更加合理、简便和高效,对离子交换法(IEX)纯化SOD的相关条件进行了探索和优化。最后确定的最佳工艺条件为:离子交换流动相为pH8.020mmol/L的Tris-HCl缓冲液,分步洗脱的条件为10%B液20min→30%B液15min→100%B液15min,最佳上样量为4mL,最大流速为150cm/h。离子交换后所得酶样比活力为4052U/mg蛋白,回收率为59.7%,纯度提高942倍。优化后的纯化工艺可以为今后SOD的纯化工艺研究和工业化生产提供科学、可靠的技术指导。

To get a reasonable,simple and efficient purification process of superoxide dismutase（SOD）from tilapia blood,the purification conditions of SOD by ion exchange chromatography（IEX）were explored and optimized.At last,the optimized purification conditions of SOD were fixed as follows：the mobile phase was pH8.0 20mmol/L Tris-HCl buffer,the stepwise elution method was 10%B、20min→30%B、15min→100%B、15min,the optimum sample volume was 4mL and the maximum flow rate was 150cm/h.The results showed that the specific enzyme activity of the purified tilapia blood SOD was 4052U/mg protein with a yield of 59.7% and a purification fold of 942 times,respectively.The optimized purification technology of SOD could provide a scientific and reliable technical guidance for future purification research and commercial process of SOD.