Open AccessStrategies to Optimize Protein Expression in E. coli
Author(s) -
Francis Dana M.,
Page Rebecca
Publication year - 2010
Publication title -
current protocols in protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.409
H-Index - 32
eISSN - 1934-3663
pISSN - 1934-3655
DOI - 10.1002/0471140864.ps0524s61
Subject(s) - escherichia coli , heterologous , protein folding , escherichia coli proteins , protein expression , heterologous expression , chaperone (clinical) , recombinant dna , biochemistry , biology , cytosol , protein engineering , protein biosynthesis , bacterial protein , computational biology , chemistry , enzyme , gene , medicine , pathology
Recombinant protein expression in Escherichia coli ( E. coli ) is simple, fast, inexpensive, and robust, with the expressed protein comprising up to 50 percent of the total cellular protein. However, it also has disadvantages. For example, the rapidity of bacterial protein expression often results in unfolded/misfolded proteins, especially for heterologous proteins that require longer times and/or molecular chaperones to fold correctly. In addition, the highly reductive environment of the bacterial cytosol and the inability of E. coli to perform several eukaryotic post‐translational modifications results in the insoluble expression of proteins that require these modifications for folding and activity. Fortunately, multiple, novel reagents and techniques have been developed that allow for the efficient, soluble production of a diverse range of heterologous proteins in E. coli . This overview describes variables at each stage of a protein expression experiment that can influence solubility and offers a summary of strategies used to optimize soluble expression in E. coli . Curr. Protoc. Protein Sci . 61:5.24.1‐5.24.29. © 2010 by John Wiley & Sons, Inc.