Tuned Escherichia coli as a host for the expression of disulfide-rich proteins

Biotechnol J. 2011 Jun;6(6):686-99. doi: 10.1002/biot.201000335. Epub 2011 May 13.


Disulfide-bond formation is a major post-translational modification and is essential for protein folding, stability, and function. This is especially true for secreted proteins, many of which possess great potential for biotechnological applications. Focusing on the use of Escherichia coli for the production of this class of proteins, we describe the mechanisms that maintain redox compartmentalization in the cell, with an emphasis on those that promote the formation and isomerization of disulfide bonds in the bacterial periplasm, while presenting parallel pathways in the eukaryotic endoplasmic reticulum. Based on these concepts, we review the use of E. coli as a cell factory for the production of heterologous disulfide-containing proteins using either peri- or cytoplasmic expression and, in particular, how these compartments can be tuned to improve the yield of correctly folded recombinant proteins. Finally, we describe a few examples of the production of small disulfide-rich proteins (protease inhibitors) to illustrate how soluble, active, and fully oxidized recombinants may be successfully obtained upon peri- or cytoplasmic expression in E. coli.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Cysteine / chemistry
  • Cysteine / metabolism
  • Cytoplasm* / genetics
  • Cytoplasm* / metabolism
  • Disulfides / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Oxidoreductases / metabolism
  • Periplasm* / genetics
  • Periplasm* / metabolism
  • Protease Inhibitors / metabolism*
  • Protein Folding
  • Protein Processing, Post-Translational / physiology*
  • Recombinant Proteins* / biosynthesis
  • Recombinant Proteins* / genetics
  • Sulfhydryl Compounds / metabolism


  • Disulfides
  • Protease Inhibitors
  • Recombinant Proteins
  • Sulfhydryl Compounds
  • Oxidoreductases
  • Cysteine