title: One-step purification and immobilization in cellulose of the GroEL apical domain fused to a carbohydrate-binding module and its use in protein refolding.
abstract: The apical domain of the chaperonin, GroEL, fused to the carbohydrate binding module type II, CBD(Cex), 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.
authors.affiliation: Departamento de Biotecnología y Bioingeniería, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Av. IPN 2508, Col. San Pedro Zacatenco, C.P. 07360, México, D. F., México.
authors.forename: Lucero A
authors.initials: LA
authors.lastname: Ramón-Luing
authors.forename: Abimael
authors.initials: A
authors.lastname: Cruz-Migoni
authors.forename: Roberto
authors.initials: R
authors.lastname: Ruíz-Medrano
authors.forename: Beatriz
authors.initials: B
authors.lastname: Xoconostle-Cázares
authors.forename: Jaime
authors.initials: J
authors.lastname: Ortega-Lopez
mesh_terms: D040143:Cellulomonas / Q000378:metabolism*; D018834:Chaperonin 60 / Q000737:chemistry* / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods*; D011485:Protein Binding; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000737:chemistry*; D013884:Thiosulfate Sulfurtransferase / Q000737:chemistry*
chemical_list: D018834:Chaperonin 60; D011956:Receptors, Cell Surface; C055213:saccharide-binding proteins; D013884:Thiosulfate Sulfurtransferase