重组水蛭素Ⅲ的分离纯化与鉴定

Purification and Identification of Recombinant Hirudin III

目的 :从分泌型重组水蛭素 Ⅲ 基因工程菌发酵液中分离纯化水蛭素 Ⅲ。 方法 :将发酵液经离心、大孔吸附树脂处理后 ,用DEAE- celluloseDE5 2离子交换层析 ,进而用HPLC反相层析。结果 :总收率达 5 0 %以上 ,纯度达 99.9% ,比活力达 90 0 0ATU/mg。结论 :分离纯化系统令人满意 ,回收率较高。

AIM:To isolate and purify recombinant hirudinⅢ,which is secreted into the culture.METHOD:A combined technique of low-temperature centrifugation,macroporous resin adsorption,DEAE-cellulose ion-exchange chromatography and RP-HPLC was developed to extract and purify the recombinant hirudinⅢ.RESULT:The recovery of extraction and purification was over 50% and the purity of hirudin was over 99%. Its antithrombin activity was 9 000 ATU/mg.