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	Immobilized beta-cyclodextrin polymer coupled to agarose gel properly refolding recombinant Staphylococcus aureus elongation factor-G in combination with detergent micelle
	Li, JJ; Venkataramana, M; Sanyal, S; Janson, JC; Su, ZG
刊名	PROTEIN EXPRESSION AND PURIFICATION
	2006
卷号	45期号:1页码:72-79
关键词	protein refolding chaperone EF-G beta-cyclodextrin polymer immobilization
ISSN号	1046-5928
其他题名	Protein Expr. Purif.
中文摘要	A novel artificial chaperone system using a combination of interactions between the unfolded protein, a detergent and a chromatographic column packed with immobilized P-cyclodextrin (P-CD) polymer coupled to an agarose gel, was introduced to refold recombinant Staphylococcus aureus elongation factor-G (EF-G). Pre-mixing of 10% Triton X-100 and unfolded EF-G at 24 mg/ml followed by a 20-fold dilution into refolding buffer led to successful capturing of EF-G by Triton X-100 resulting in formation of a detergent-protein complex at 1.2 mg/ml of final protein concentration. The complex was subsequently applied to the immobilized beta-CD polymer column resulting in correct refolding of EF-G at a concentration of 530 mu g/ml with 99% mass recovery. Detergent concentrations above critical micelle concentration were required for efficient capturing of EF-G at high protein concentration. Other detergents with hydrophile-lipophile-Balance values similar to that of Triton X-100 (Triton N-101, Noindet P40 (NP40), and Berol 185) also produced similar result. Soluble polymerized P-CD was more efficient than the monomer to remove the detergent from the protein complex in a batch system. Immobilized P-CD polymer column further improved the capability of detergent removal and was able to prevent aggregation that occurred with the addition of soluble P-CD polymer at high protein concentration in the batch system. The mechanism for this system-assisted refolding was tentatively interpreted: the released protein could correctly refold in an enclosed hydrophilic environment provided by the integration of matrix and P-CD polymer, and thus avoided aggregation during detergent removal. (C) 2005 Elsevier Inc. All rights reserved.
英文摘要	A novel artificial chaperone system using a combination of interactions between the unfolded protein, a detergent and a chromatographic column packed with immobilized P-cyclodextrin (P-CD) polymer coupled to an agarose gel, was introduced to refold recombinant Staphylococcus aureus elongation factor-G (EF-G). Pre-mixing of 10% Triton X-100 and unfolded EF-G at 24 mg/ml followed by a 20-fold dilution into refolding buffer led to successful capturing of EF-G by Triton X-100 resulting in formation of a detergent-protein complex at 1.2 mg/ml of final protein concentration. The complex was subsequently applied to the immobilized beta-CD polymer column resulting in correct refolding of EF-G at a concentration of 530 mu g/ml with 99% mass recovery. Detergent concentrations above critical micelle concentration were required for efficient capturing of EF-G at high protein concentration. Other detergents with hydrophile-lipophile-Balance values similar to that of Triton X-100 (Triton N-101, Noindet P40 (NP40), and Berol 185) also produced similar result. Soluble polymerized P-CD was more efficient than the monomer to remove the detergent from the protein complex in a batch system. Immobilized P-CD polymer column further improved the capability of detergent removal and was able to prevent aggregation that occurred with the addition of soluble P-CD polymer at high protein concentration in the batch system. The mechanism for this system-assisted refolding was tentatively interpreted: the released protein could correctly refold in an enclosed hydrophilic environment provided by the integration of matrix and P-CD polymer, and thus avoided aggregation during detergent removal. (C) 2005 Elsevier Inc. All rights reserved.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
研究领域[WOS]	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]	ARTIFICIAL CHAPERONE ; MOLECULAR CHAPERONES ; INCLUSION-BODIES ; PROTEIN ; RENATURATION ; LYSOZYME
收录类别	SCI
原文出处	://WOS:000234521100010
语种	英语
WOS记录号	WOS:000234521100010
公开日期	2013-10-24
内容类型	期刊论文
版本	出版稿
源URL	[http://ir.ipe.ac.cn/handle/122111/3991]
专题	过程工程研究所_研究所（批量导入）
作者单位	1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100080, Peoples R China 2.Uppsala Univ, BMC, Dept Cell & Mol Biol, SE-75124 Uppsala, Sweden 3.Uppsala Univ, BMC, Dept Surface Biotechnol, SE-75123 Uppsala, Sweden
推荐引用方式 GB/T 7714	Li, JJ,Venkataramana, M,Sanyal, S,et al. Immobilized beta-cyclodextrin polymer coupled to agarose gel properly refolding recombinant Staphylococcus aureus elongation factor-G in combination with detergent micelle[J]. PROTEIN EXPRESSION AND PURIFICATION,2006,45(1):72-79.
APA	Li, JJ,Venkataramana, M,Sanyal, S,Janson, JC,&Su, ZG.(2006).Immobilized beta-cyclodextrin polymer coupled to agarose gel properly refolding recombinant Staphylococcus aureus elongation factor-G in combination with detergent micelle.PROTEIN EXPRESSION AND PURIFICATION,45(1),72-79.
MLA	Li, JJ,et al."Immobilized beta-cyclodextrin polymer coupled to agarose gel properly refolding recombinant Staphylococcus aureus elongation factor-G in combination with detergent micelle".PROTEIN EXPRESSION AND PURIFICATION 45.1(2006):72-79.

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