title: Tuned Escherichia coli as a host for the expression of disulfide-rich proteins.
abstract: 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.
authors.affiliation: Cátedra de Inmunología, Facultad de Química-Facultad de Ciencias, Instituto de Higiene, Universidad de la República, Montevideo, Uruguay.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Salinas
authors.forename: Leonardo
authors.initials: L
authors.lastname: Pellizza
authors.forename: Mariana
authors.initials: M
authors.lastname: Margenat
authors.forename: Martín
authors.initials: M
authors.lastname: Fló
authors.forename: Cecilia
authors.initials: C
authors.lastname: Fernández
mesh_terms: D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism; D019897:Periplasm / Q000235:genetics / Q000378:metabolism; D011480:Protease Inhibitors / Q000378:metabolism*; D017510:Protein Folding; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: D004220:Disulfides; D011480:Protease Inhibitors; D011994:Recombinant Proteins; D013438:Sulfhydryl Compounds; D010088:Oxidoreductases; D003545:Cysteine