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- Title
Production of functional inclusion bodies in endotoxin-free Escherichia coli.
- Authors
Rueda, Fabián; Cano-Garrido, Olivia; Mamat, Uwe; Wilke, Kathleen; Seras-Franzoso, Joaquin; García-Fruitós, Elena; Villaverde, Antonio
- 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.
- Subjects
ESCHERICHIA coli; ENDOTOXINS; MOLECULAR cloning; RECOMBINANT proteins manufacturing; BIOTECHNOLOGICAL microorganisms
- Publication
Applied Microbiology & Biotechnology, 2014, Vol 98, Issue 22, p9229
- ISSN
0175-7598
- Publication type
Article
- DOI
10.1007/s00253-014-6008-9