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.
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Acknowledgments
We are indebted to the Protein Production Platform (CIBER-BBN - UAB) for helpful technical assistance and for protein production and purification services (http://www.ciber-bbn.es/en/programas/89-plataforma-de-produccion-de-proteinas-ppp). The authors acknowledge the financial support granted to E.G.F. from INIA, (MINECO, RTA2012-00028-C02-02) Spain, and to A.V. from Agència de Gestió d’Ajuts Universitaris i de Recerca (2014SGR-132) and from the Centro de Investigación Biomédica en Red (CIBER) de Bioingeniería, Biomateriales y Nanomedicina financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. F.R. was supported by a predoctoral fellowship (Beca de Formación Doctoral "Francisco José de Caldas", Convocatoria 512-2010 de Colciencias) and O.C-G received a PhD fellowship from MECD. A.V. has been distinguished with an ICREA ACADEMIA Award.
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Rueda, F., Cano-Garrido, O., Mamat, U. et al. Production of functional inclusion bodies in endotoxin-free Escherichia coli . Appl Microbiol Biotechnol 98, 9229–9238 (2014). https://doi.org/10.1007/s00253-014-6008-9
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DOI: https://doi.org/10.1007/s00253-014-6008-9