Unit

UNIT 5.24 Strategies to Optimize Protein Expression in E. coli

  1. Dana M. Francis,
  2. Rebecca Page

Published Online: 1 AUG 2010

DOI: 10.1002/0471140864.ps0524s61

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Francis, D. M. and Page, R. 2010. Strategies to Optimize Protein Expression in E. coli. Current Protocols in Protein Science. 61:5.24:5.24.1–5.24.29.

Author Information

  1. Brown University, Providence, Rhode Island

Publication History

  1. Published Online: 1 AUG 2010
  2. Published Print: AUG 2010

Abstract

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.

Keywords:

  • protein expression;
  • E. coli;
  • fusion proteins;
  • proteases;
  • heterologous protein;
  • purification tags;
  • expression tags;
  • expression strains and vectors;
  • folded protein;
  • active protein