猪牛血块超氧化物歧化酶提取工艺研究(2)——热变温度和时间对SOD提取的影响及最佳工艺条件选择

Studies on the Extraction Technique of Superoxide Dismutase from Pig and Bovine Blood Block(2)——The Influence of Thermal Denaturation Temperature and Time on Extraction of SOD,and the Selection of Optimal Technique Condition

目的:重点研究热变温度、热变时间对SOD分离提取的影响,确定最优工艺参数。方法:采用正交试验设计。结果:热变温度:牛血65～70℃,猪血60～65℃;热变时间:牛血15～20min,猪血10～15min。由正交试验得出牛血的最优组合为A2B2C2,即溶膜剂加量15%,热变温度65℃,热变时间15min;猪血最佳组合为A1B2C2,即溶膜剂加量20%,热变温度60℃,热变时间15min。结论:热变是猪、牛血SOD提取的关键,按此工艺纯化的SOD活性在3000U/mg蛋白以上。

Objective:To study the effect of thermal denaturation temperature and thermal denaturation time on extraction of SOD,and to define its optimal technique parameter.Method:Orthogonal experiment design was adopted.Results:From synthetical analysis of the SOD activity and yield productive,the thermal denaturation temperature of bovine blood block is 65～70 ℃,and the thermal denaturation temperature of pig blood block is 60～65 ℃;thermal denaturation time of bovine blood block is 15～20 min,thermal denaturation time of pig blood block is 10～15 min.The optimal combination of orthogonal experiment,bovine blood block is A2B2C2,that is,membranelytic agent addition is 15%,the thermal denaturation temperature is 65 ℃;The thermal denaturation time is 15 min;The optimal combination of pig blood is A1B1C2,that is,membranelytic agent is 20%,the thermal denaturation temperature is 60 ℃,the thermal denaturation time is 15 min.Conclusion:Thermal denaturation is one of the most importance in extracting SOD from pig and bovine blood block,higher purity SOD,whose activity exceeds 3 000 U/mg protein was reached by this technique.