利用膨胀床吸附技术单步纯化分子伴侣—GroEL

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利用膨胀床吸附技术单步纯化分子伴侣—GroEL

佟晓冬; 杨征; 董晓燕; 孙彦

Department of Biochemical Engineering, School of Chemical Engineering and Technology,
Tianjin University, Tian-jin 300072, China

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2003-08-28 发布日期:2003-08-28
通讯作者: 佟晓冬

Single-step Purification of Molecular Chaperone GroEL by Expanded Bed Chromatography

TONG Xiaodong; YANG Zheng; DONG Xiaoyan; SUN Yan

Department of Biochemical Engineering, School of Chemical Engineering and Technology,
Tianjin University, Tian-jin 300072, China

Received:1900-01-01 Revised:1900-01-01 Online:2003-08-28 Published:2003-08-28
Contact: TONG Xiaodong

摘要/Abstract

摘要： Expanded bed adsorption (EBA) is an integrative downstream processing technique for the
purificationof biological substances directly from unclarified feedstock. In this study,
molecular chaperone GroEL, an importantprotein folding helper both in vivo and in vitro,
was purified by the single-step EBA technique from the unclarifiedhomogenate of recombinant
E. coli cells. Compared with packed bed adsorption, the EBA technique provideda single-step
approach to yield an electrophoretic purity of GroEL. After the homogenate loading and
columnwashing 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-bedmode in the purification of GroEL from
cell disruptate.

关键词: expanded bed adsorption;molecular chaperone;GroEL;purification

Abstract: Expanded bed adsorption (EBA) is an integrative downstream processing technique for the
purificationof biological substances directly from unclarified feedstock. In this study,
molecular chaperone GroEL, an importantprotein folding helper both in vivo and in vitro,
was purified by the single-step EBA technique from the unclarifiedhomogenate of recombinant
E. coli cells. Compared with packed bed adsorption, the EBA technique provideda single-step
approach to yield an electrophoretic purity of GroEL. After the homogenate loading and
columnwashing 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-bedmode in the purification of GroEL from
cell disruptate.

Key words: expanded bed adsorption, molecular chaperone, GroEL, purification

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]. CIESC Journal.

TONG Xiaodong, YANG Zheng, DONG Xiaoyan, SUN Yan. Single-step Purification of Molecular Chaperone GroEL by Expanded Bed Chromatography[J]. .

参考文献

[1] Ellis, R.J., van der Vries, S.M. "Molecular chaperones",Annu. Rev. Biochem., 60, 321
(1991).
[2] Fisher, M.T., "Promotion of the in vitro renaturation of dodecametric glutamine
synthetase from Escherichia coli in the presence of GroEL (chaperonin-60) and ATP",
Biochemistry, 31, 3955 (1992).
[3] Lemos, J.A.C., Buene, R.A., Castro, A.C.D., "Molecular cloning, purification and
immunological responses of recombinants GroEL and DnaK from Streptococcus pyogenes", FEMS
Immunology and Medical Microbiology, 28,121 (2000).
[4] Blennow, A., Surin, B.P., Ehring, H., McLennan, N.F.,Spangfort, M. D., "Isolation and
biochemical characterization of highly purified Escherichia coli molecular chaperone Cpn60
(GroEL) by affinity chromatography and ureainduced monomerization", Biochimica et
Biophysica Acta,1252, 69 (1995).
[5] Jenkins, A.J., March, J.B., Oliver, I.R., Masters, M., "DNA fragment containing the
GroE genes can suppress mutations in Escherichia coli dnaA genes", Mol. Gen. Genet.,
202,446 (1986).
[6] Bradford, M.M., "A rapid and sensitive method for the quantitation of microgram
quantities of protein utilizing t he principle of protein-dye", Anal. Biochem., 72, 248
(1976).
[7] Yang, H., "Immobilized chaperonin GroEL facilitates the refolding of denatured
lysozyme", Master thesis, Tianjin University, Tianjin (1999). (in Chinese)
[8] Amersham Pharmacia Biotech., "Expanded bed adsorption-principles and methods",
(Handbook), Amersham Pharmacia Biotech. Uppsala, Sweden (2000).
[9] Hjorth, R., "Expanded bed adsorption: Elution in expanded bed mode", Bioseparation, 8,
1 (1999).
[10] Chase, H.A., Draeger, N.M., "Expanded-bed adsorption of proteins using ion-
exchangers", Separation Science and Technology, 27, 2021 (1992).
[11] Xu, Z., Chen, X.M, Zheng, W.D. Yang, S.L., Zhu,D.X., "The expression, purification and
characterization of GroEL and GroES proteins in E. coli.", Journal of Nanjin University, 33
(1), 67 (1997). (in Chinese)
[12] Hu, H.B., Yao, S.J., Lin, D.Q., Zhu, Z.Q, "Adsorption of protein in the expanded bed",
Chinese J. Chem. Eng., 8,230 (2000).

利用膨胀床吸附技术单步纯化分子伴侣—GroEL

Single-step Purification of Molecular Chaperone GroEL by Expanded Bed Chromatography

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