蚯蚓纤溶酶的分离纯化及其动力学性质研究

Purification and kinetic characteristics of fibrinolytic enzymes from earthworm

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

Objective:To achieve an effective method of purifying earthworm fibrinolytic enzymes (EFE), and study the characteristics of EFE as well.Methods:The procedures of purification included buffer extraction, selective thermal denaturation and chromatographies on EFE inhibitor 1.6 hexamethylediamine (HD) Sepharose 4B and DEAE Sepharose. Results and conclusion:Strong fibrinolytic components (EFE F 1、 2, EFE F 3) were purified. For EFE F 3, Km of S 2288 is 0.01mmol/L, Km of Chromozym P is 0.02 mmol/L, Ki of HD is 1mmol/L; it is quite stable to heat treatment and 4mol/L Urea.