Abstract
The apical domain of the chaperonin, GroEL, fused to the carbohydrate binding module type II, CBDCex, of Cellulomonas fimi, was expressed in Escherichia coli. The recombinant protein, soluble or from inclusion bodies, was directly purified and immobilized in microcrystalline cellulose particles or cellulose fabric membranes. Assisted refolding of rhodanese by the immobilized mini-chaperone showed a two-fold improvement as compared to a control. Using chromatographic refolding, 35% of rhodanese activity was recovered in only 5 min (mean residence time) as compared to 17% for spontaneous refolding. This mini-chaperone immobilized in cellulose could be a cost-efficient method to refold recombinant proteins expressed as inclusion bodies.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
MM Altamirano R Golbik R Zahn AM Buckle AR Fersht (1997) ArticleTitleRefolding chromatography with immobilized mini-chaperones Proc. Natl. Acad. Sci. USA 94 3576–3578 Occurrence Handle10.1073/pnas.94.8.3576 Occurrence Handle1:CAS:528:DyaK2s**s1amtbY%3D Occurrence Handle9108018
AC Clark E Hugo C Frieden (1996) ArticleTitleDetermination of regions in the dihydrofolate reductase structure that interact with the molecular chaperonin GroEL Biochemistry 35 5893–5901 Occurrence Handle1:CAS:528:DyaK28**sV2jsb8%3D Occurrence Handle8639551
YG Gao YX Guan SJ Yao MG Cho (2003) ArticleTitleOn-column refolding of recombinant human interferon-γ with an immobilized chaperone fragment Biotechnol. Prog. 19 915–920 Occurrence Handle10.1021/bp025775l Occurrence Handle1:CAS:528:DC%2BD3s**sValu7s%3D Occurrence Handle12790657
R Golbik R Zahn SE Harding AR Fersht (1998) ArticleTitleThermodynamic stability and folding of GroEL minichaperones J. Mol. Biol. 276 505–515 Occurrence Handle10.1006/jmbi.1997.1538 Occurrence Handle1:CAS:528:DyaK1cXhs1Sgtrs%3D Occurrence Handle9512719
A Jungbauer W Kaar R Schlegl (2004) ArticleTitleFolding and refolding of proteins in chromatographic beds Curr. Opin. Biotechnol. 15 487–494 Occurrence Handle10.1016/j.copbio.2004.08.009 Occurrence Handle1:CAS:528:DC%2BD2cXotFCrs7w%3D Occurrence Handle15464382
M Li ZG Su JC Janson (2004) ArticleTitleIn vitro protein refolding by chromatographic procedures Protein Exp. Purif. 33 1–10
J Martin FU Hartl (1997) ArticleTitleChaperone-assisted protein folding Curr. Opin. Struct. Biol. 7 41–52 Occurrence Handle10.1016/S0959-440X(97)80006-1 Occurrence Handle1:CAS:528:DyaK2sXhtleqs74%3D Occurrence Handle9032064
JA Mendoza E Rogers GH Lorimer PM Horowitz (1991) ArticleTitleChaperonins facilitate the In vitro folding of monomeric mitochondrial rhodanese J. Biol. Chem. 266 13044–13049 Occurrence Handle1:CAS:528:DyaK3MXlslyis70%3D Occurrence Handle1677004
AR Middelberg (2002) ArticleTitlePreparative protein refolding Trends Biotechnol. 20 437–443 Occurrence Handle10.1016/S0167-7799(02)02047-4 Occurrence Handle1:CAS:528:DC%2BD38XmvVWmtLY%3D Occurrence Handle12220907
E Ong NR Gilkes RC Miller RAJ Warren DG Kilburn (1993) ArticleTitleThe Cellulose-Binding Domain (CBDCex) of an exoglucanase from Cellulomonas fimi: production in Escherichia Coli and characterization of the polypeptide Biotechnol. Bioeng. 42 401–409 Occurrence Handle10.1002/bit.260420402 Occurrence Handle1:CAS:528:DyaK3sXltlOnt74%3D
CH Schein MHM Noteborn (1988) ArticleTitleFormation of soluble recombinant proteins in Escherichia Coli is favored by lower growth temperature Bio/Technology 6 291–294 Occurrence Handle10.1038/nbt0388-291 Occurrence Handle1:CAS:528:DyaL1cXkt1Wmur0%3D
BH Sorbo (1955) Rhodanese SP Colowick NO Kaplan (Eds) Methods in Enzymology: Preparation and Assay of Enzymes Academic Press San Diego 334–337
A Tejeda-Mansir RM Montesinos I Magana-Plaza R Guzman (2003) ArticleTitleBreakthrough performance of stacks of dye-cellulosic fabric in affinity chromatography of lysozyme Bioprocess Biosyst. Eng. 25 235–242 Occurrence Handle1:CAS:528:DC%2BD3s**tVWqsbk%3D Occurrence Handle14505002
P Tomme A Boraston B McLean J Kormos A Creagh K Sturch NR Gilkes CA Haynes RAJ Warren DG Kilburn (1998) ArticleTitleCharacterization and affinity applications of cellulose-binding domains J. Chromatogr. B 715 283–296 Occurrence Handle1:CAS:528:DyaK1cXlvFektLY%3D
M Wood Thomas (1988) Preparation of crystalline, amorphous, and dyed cellulase substrates WA Wood ST Kellogg (Eds) Methods in Enzymology: Biomass Academic Press San Diego 19–25
R Zahn AM Buckle S Perrett CM Johnson FJ Corrales R Golbik AR Fersht (1996) ArticleTitleChaperone activity and structure of monomeric polypeptide binding domains of GroEL Proc. Natl. Acad. Sci. USA 93 15024–15029 Occurrence Handle10.1073/pnas.93.26.15024 Occurrence Handle1:CAS:528:DyaK2sXmslOr Occurrence Handle8986757
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ramón-Luing, L.A., Cruz-Migoni, A., Ruíz-Medrano, R. et al. One-step Purification and Immobilization in Cellulose of the GroEL Apical Domain Fused to a Carbohydrate-binding Module and its use in Protein Refolding. Biotechnol Lett 28, 301–307 (2006). https://doi.org/10.1007/s10529-005-5714-x
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10529-005-5714-x