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目的 :建立一种高效的蚯蚓纤溶酶 (EFE)的分离纯化技术 ,并研究其特性。方法 :蚯蚓经组织匀浆、缓冲液抽提与选择性热变性 ,再经抑制剂 1,6 -己二胺 (HD)偶联的 Sepharose4B亲和层析 ,DEAE-Sepharose离子交换层析。结果及结论 :分离纯化得到具有强烈纤溶活性的酶组分 (EFE- F1 、2 ,EFE- F3)。其中 EFE- F3对合成底物 S- 2 2 88和 Chrom ozym P的 Km分别是 0 .0 1mm ol/L和 0 .0 2 mm ol/L ,HD的 Ki为1mmol/L;该酶热稳定性强 ,能抗 4mol/L 脲
Objective : To establish a highly efficient separation and purification technique of limulus plasmin (EFE) and to study its characteristics. METHODS: Rats were homogenized, buffer extracted and selectively heat denatured, and then treated with 1,6-hexanediamine (HD) coupled with Sepharose4B affinity chromatography and DEAE-Sepharose ion exchange chromatography. Results and conclusions: The isolated and purified enzyme components (EFE-F1,2,EFE-F3) have strong fibrinolytic activity. The Km of EFE-F3 for synthetic substrates S- 2 88 and Chrom ozym P were 0. 0 1 mm ol/L and 0.02 mm ol/L, respectively, and Ki for HD was 1 mmol/L; the enzyme was thermally stable. Strong, resistant to 4mol/L urea