A fast and simple method to eliminate Cpn60 from functional recombinant proteins produced by E. coli Arctic Express.
2015
BELVAL, Lorène | Marquette, Arnaud | Mestre Artigues, Pedro-Felipe | Piron, Marie-Christine | Demangeat, Gerard | Merdinoglu, Didier | Chich, Jean-Francois
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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