发酵液中重组水蛭素的分离纯化

Isolation and Purification of Recombinant Hirudin-2 From Fermentation Medium

摘要: 目的：建立毕赤酵母(pichia)高密度表达的重组水蛭素(recombinant hirudin-2,rHV2)分离纯化工艺。方法：高密度培养重组毕赤酵母并诱导表达水蛭素至胞外，发酵液经超滤、透析、离子交换层析。分离纯化rHV2目的蛋白；采用SDS-PAGE和抗凝血酶活力分别检测目的蛋白纯度和活性。结果：纯化样品经SDS-PAGE电泳检测为单一条带，重组水蛭素比活性为6000ATU／mg，纯化收率达38．6％。结论：超滤、透析和离子交换层析可用于发酵液中大规模分离纯化重组水蛭素。

Abstract: AIM:The purpose is to establish the isolation and purification process of recombinant hirudin-2(rHV 2 ) expressed in pichia. METHOD: Recombinant pichia was cultured with high cell density and induced to express rHV 2 to culture medium,rHV 2 was