中草药桃仁纤溶酶的亲和层析分离

Affinity Chromatography of Fibrinolytic Enzyme from Peach Kernel

目的分离出高效、特异、安全、不良反应较小的纤溶酶,为溶栓药物的研究奠定基础。方法利用硫铵沉淀法提取出桃仁纤溶酶,测定其比活力,并比较不同蛋白酶抑制剂对桃仁纤溶酶活性的抑制作用,采用亲和层析法对桃仁纤溶酶进行分离纯化。结果亲和层析法分离得到了纤溶酶活性单一组分,比活力为89.08 U.mg?1,比纯化前提高了7.37倍,该组分属于丝氨酸蛋白酶家族。结论利用大豆胰蛋白酶抑制剂作为配基,采用亲和层析法分离桃仁纤溶酶可以得到单一活性成分,且比活力相对提高。

OBJECTIVE To separate the fibrinolytic enzyme which is highly effective,specific,security and few side effects,to lay the foundation for the thrombolytic drugs.METHODS The crude fibrinolytic enzyme from peach kernel was precipitated with saturated ammonium sulfate and purified by affinity chromatography.The specific activity of the crude fibrinolytic enzyme was detected.Compared the inhibitory action of different proteinase inhibitors,separated and purified the crude fibrinolytic enzyme with the method of affinity chromatography.RESULTS A single active component was isolated from crude fibrinolytic enzyme with the method of affinity chromatography.The specific activity of the component was 89.08 U.mg？1,which was 7.37 times higher than that before purified.This component belongs to the serine proteinase family.CONCLUSION Using the inhibitor of soybean trypsin as a ligand,a single active component could be obtained by affinity chromatography.And the specific activity increased relatively.