| 发酵液中重组水蛭素的分离纯化 |
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| 引用本文： | 发酵液中重组水蛭素的分离纯化[J]. 中国药科大学学报, 2003, (3): 74-77. |
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| 作者姓名： | 邵雷 周长林 范维 赵明龙 |
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| 作者单位： | 1. 中国药科大学微生物学教研室,南京,210009
2. 上海美通生物科技有限公司,上海,200235 |
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| 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。
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| 关 键 词： | 重组水蛭素 凝血酶抑制剂 纯化 |
| 文章编号： | 1000-5048(2003)03-0268-04 |
| 修稿时间： | 2002-08-23 |
Isolation and Purification of Recombinant Hirudin-2 From Fermentation Medium |
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| Isolation and Purification of Recombinant Hirudin-2 From Fermentation Medium[J]. Journal of China Pharmaceutical University, 2003, (3): 74-77. |
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| Authors: | SHAO Lei ZHOU Chang-Lin FAN Wei ZHAO Ming-Long School of Bio-Pharmaceutics China Pharmaceutical University Nanjing China Shanghai General Scientific Biotechnology Co. Shanghai China |
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| Affiliation: | SHAO Lei 1,ZHOU Chang-Lin 1,FAN Wei 1,ZHAO Ming-Long 2 1School of Bio-Pharmaceutics,China Pharmaceutical University,Nanjing 210009,China 2 Shanghai General Scientific Biotechnology Co.,Shanghai 200235,China |
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| 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 purified from fermentation medium by ultrafiltration,dialysis, ion exchange chromatography and analyzeed by SDS-PAGE and antithrombin assay for the detection of purity and activity of rHV 2 . RESULT:SDS-PAGE demonstrated that the purified rHV 2 showed a single band with the specific activity of 6000 ATU/mg and recovery of 38.6% respectively. CONCLUSION:This technology can be applied to the purification of rHV 2 from fermentation medium on large scale. |
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| Keywords: | Recombinant hirudin-2 Thrombin-specific inhibitor Purification |
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