低分子质量蚯蚓纤溶酶的获得及其活性研究

Study on Purification and Activity of Earthworm Fibrinolytic Enzyme (EFE) with Lower M_r

分离纯化出两种M_r较小的蚯蚓纤溶酶组分,并进行了部分氨基酸序列测定及初步的药理活性研究。以新鲜赤子爱胜蚓为原料,通过超滤、DEAE-Sepharose fast flow离子交换层析及Sephadex G75凝胶层析,得到两电泳纯组分,M_r分别为20 k与9 k,纤维平板法测得其比活分别为96 IU/mg与10 IU/mg,部分氨基酸序列测定结果显示20 k组分与已知蚯蚓纤溶酶gi|157931564有很强的同源性,9 k组分与水蛭的神经血蓝蛋白具同源性。药理实验表明,两组分均能促进人脐静脉内皮细胞(huVEC)分泌t-PA。抑菌实验表明,两组分及蚯蚓水提液中M_r 4～10 k部分均具有广泛抗菌谱且抑菌效果4～10 k部分>F2>F1。

The two active components F1 and F2 were isolated from earthworm Eisenia foelide by ultrafiltration, ion exchange and gel chromatography. Both of them displayed single band on SDS-PAGE with Mr 20k and 9k, respectively. Analysis of partial amino acid sequence indicated homology with gi1157931564 for F1 and Neurohemerythrin for F2. Their thrombolytic activity in vivo and in vitro was also measured. Fibrin specific activity was 96IU/mg（F1） and 10 IU/mg（F2）. They activated t-PA activity in huVEC cells. They also showed the broad antimicrobial activity against Gram-positive and Gram-negative bacteria.