| 利用膨胀床吸附技术单步纯化分子伴侣—GroEL |
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| 引用本文： | 佟晓冬,杨征,董晓燕,孙彦.利用膨胀床吸附技术单步纯化分子伴侣—GroEL[J].中国化学工程学报,2003,11(4):460-463. |
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| 作者姓名： | 佟晓冬 杨征 董晓燕 孙彦 |
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| 作者单位： | DepartmentofBiochemicalEngineering,SchoolofChemicalEngineeringandTechnology,TianjinUniversity,Tian-jin300072,China |
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| 基金项目： | Supported by the National Natural Science Foundation of China (No. 20025617). |
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| 摘 要： | Expanded bed adsorption (EBA) is an integrative downstream processing technique for the purification of biological substances directly from unclarified feedstock. In this study, molecular chaperone GroEL, an important protein folding helper both in vivo and in vitro, was purified by the single-step EBA technique from the unclarified homogenate of recombinant E. coli cells. Compared with packed bed adsorption, the EBA technique provided a single-step approach to yield an electrophoretic purity of GroEL. After the homogenate loading and column washing in the expanded bed mode, the GroEL protein was recovered by stepwise salt-gradient elution in packed-bed or expanded-bed modes, respectively. The expanded-bed elution mode was found as efficient as the packed-bed mode in the purification of GroEL from cell disruptate.
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| 关 键 词： | 膨胀床 吸附技术 纯化 分子伴侣 GroEL 分离 |
| 修稿时间： | |
Single-step Purification of Molecular Chaperone GroEL by Expanded Bed Chromatography |
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| TONG Xiaodong,YANG Zheng,DONG Xiaoyan,SUN Yan.Single-step Purification of Molecular Chaperone GroEL by Expanded Bed Chromatography[J].Chinese Journal of Chemical Engineering,2003,11(4):460-463. |
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| Authors: | TONG Xiaodong YANG Zheng DONG Xiaoyan SUN Yan |
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| Affiliation: | Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tian-jin 300072, China |
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| Abstract: | Expanded bed adsorption (EBA) is an integrative downstream processing technique for the purification of biological substances directly from unclarified feedstock. In this study, molecular chaperone GroEL, an important protein folding helper both in vivo and in vitro, was purified by the single-step EBA technique from the unclarified homogenate of recombinant E. coli cells. Compared with packed bed adsorption, the EBA technique provided a single-step approach to yield an electrophoretic purity of GroEL. After the homogenate loading and column washing in the expanded bed mode, the GroEL protein was recovered by stepwise salt-gradient elution in packed-bed or expanded-bed modes, respectively. The expanded-bed elution mode was found as efficient as the packed-bed mode in the purification of GroEL from cell disruptate. |
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| Keywords: | expanded bed adsorption molecular chaperone GroEL purification |
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