正交试验法优选蚓激酶提取工艺研究

Study on Optimum Preparation Process of Earthworm Fibrinolytic Enzyme by Orthogonal Design

目的:优选蚓激酶提取工艺参数。方法:以赤子爱胜蚓为原料,通过正交试验法,以酶的活性为指标,考察缓冲液p H值、第一次盐析和第二次盐析的饱和度对提取效果的影响,得到蚓激酶粗品经凝胶层析、离子交换层析、透析、超滤等工序制备高活性蚓激酶。结果:在缓冲液p H7.5,第一次盐析硫酸胺饱和度为40%,二次饱和度为80%时,蚓激酶比活达3327 u/mg。结论:该提取工艺稳定性好,酶活性高。

Objective To select the technological parameters for extracting earthworm fibrinolytic enzyme. Methods Conditions for the extraction were studied by orthogonal design, and the specific activity of earthworm fibrinolytic enzyme were measured. The effects of buffer pH, the first and the second salting out saturation on the extraction efficiency were investigated. Extraction of earthworm fi- brinotytie enzyme crude from eisenia foetida, which was purified by gel chromatography, ion exchange chromatography, dialysis, ultra- filtration and other processes to prepare highly active earthworm fibrinolytic enzyme. Results The buffer of pH 7.5, he first saturation of 40% and secondary saturation of 80%, and the highest specific activity of crude was 3327 u/mg. Conclusion The extraction process is stable and the enzyme activity is high.