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Updated: Apr 17, 2026

Overexpressing and Purifying a Toxic Nuclease from Escherichia coli
Published on: August 29, 2025
A fast and simple method to eliminate Cpn60 from functional recombinant proteins produced by E. coli Arctic Express
Lorène Belval1, Arnaud Marquette2, Pere Mestre1
1INRA, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68021 Colmar, France; Université de Strasbourg, UMR 1131 Santé de la Vigne et Qualité du Vin, F-68000 Colmar, France.
Abstract:
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.

