A fast and simple method to eliminate Cpn60 from functional recombinant proteins produced by E. coli Arctic Express.

A frequent problem of recombinant protein production is their insolubility. To address this issue, engineered Escherichiacoli strains like Arctic Express that produce an exogenous chaperone facilitating protein folding, have been designed. A drawback is the frequent contamination of the protein by c...

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Bibliographic Details
Published in:Protein Expression and Purification
Main Authors: BELVAL, Lorène, Marquette, Arnaud, Mestre Artigues, Pedro-Felipe, Piron, Marie-Christine, Demangeat, Gerard, Merdinoglu, Didier, Chich, Jean-Francois
Format: Article in Journal/Newspaper
Language:English
Published: 2015
Subjects:
Arctic Express;Cpn60;IMAC;Recombinant protein
Arctic
Online Access:http://prodinra.inra.fr/ft/6BD75AAF-823E-462F-A69C-48EE0E6AA5A7
http://prodinra.inra.fr/record/304861
https://doi.org/10.1016/j.pep.2015.01.009
Description
Summary:A frequent problem of recombinant protein production is their insolubility. To address this issue, engineered Escherichiacoli strains like Arctic Express that produce an exogenous chaperone facilitating protein folding, have been designed. A drawback is the frequent contamination of the protein by chaperones. A simple method, using urea at a sub-denaturing concentration, allows unbinding of Cpn60 from expressed protein. This method was successfully used to purify 2 proteins, an enzyme and a viral protein. The enzyme was fully active. The nature of interaction forces between enzyme and Cpn60 was investigated. The method is likely applicable to purify other proteins.

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