A fast and simple method to eliminate Cpn60 from functional recombinant proteins produced by E. coli Arctic Express.
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 contaminat...
Published in: | Protein Expression and Purification |
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Main Authors: | , , , , , , |
Other Authors: | , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
HAL CCSD
2015
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Subjects: | |
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 |
Summary: | 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. |
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