title: Production of functional inclusion bodies in endotoxin-free Escherichia coli.
abstract: Escherichia coli is the workhorse for gene cloning and production of soluble recombinant proteins in both biotechnological and biomedical industries. The bacterium is also a good producer of several classes of protein-based self-assembling materials such as inclusion bodies (IBs). Apart from being a relatively pure source of protein for in vitro refolding, IBs are under exploration as functional, protein-releasing materials in regenerative medicine and protein replacement therapies. Endotoxin removal is a critical step for downstream applications of therapeutic proteins. The same holds true for IBs as they are often highly contaminated with cell-wall components of the host cells. Here, we have investigated the production of IBs in a recently developed endotoxin-free E. coli strain. The characterization of IBs revealed this mutant as a very useful cell factory for the production of functional endotoxin-free IBs that are suitable for the use at biological interfaces without inducing endotoxic responses in human immune cells.
authors.affiliation: Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Cerdanyola del Vallès, Spain.
authors.forename: Fabián
authors.initials: F
authors.lastname: Rueda
authors.forename: Olivia
authors.initials: O
authors.lastname: Cano-Garrido
authors.forename: Uwe
authors.initials: U
authors.lastname: Mamat
authors.forename: Kathleen
authors.initials: K
authors.lastname: Wilke
authors.forename: Joaquin
authors.initials: J
authors.lastname: Seras-Franzoso
authors.forename: Elena
authors.initials: E
authors.lastname: García-Fruitós
authors.forename: Antonio
authors.initials: A
authors.lastname: Villaverde
mesh_terms: D001688:Biological Products / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods; D004731:Endotoxins / Q000172:deficiency*; D004926:Escherichia coli / Q000378:metabolism*; D002479:Inclusion Bodies / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013678:Technology, Pharmaceutical / Q000379:methods*
chemical_list: D001688:Biological Products; D004731:Endotoxins; D011994:Recombinant Proteins; C044101:endotoxin, Escherichia coli