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

• 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-François
• Other Authors: Santé de la vigne et qualité du vin (SVQV), Institut National de la Recherche Agronomique (INRA)-Université de Strasbourg (UNISTRA), Institut de Chimie de Strasbourg, Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Département SPE
• Format: Article in Journal/Newspaper
• Language: English
• Published: HAL CCSD 2015
• Subjects: Arctic Express; IMAC; Cpn60; Recombinant protein; [SDV]Life Sciences [q-bio]; [SDV.BV]Life Sciences [q-bio]/Vegetal Biology; Arctic
• Online Access: https://hal.inrae.fr/hal-02635260; https://hal.inrae.fr/hal-02635260/document; https://hal.inrae.fr/hal-02635260/file/1-s2.0-S1046592815000108-main_1.pdf; https://doi.org/10.1016/j.pep.2015.01.009

International audience 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.