Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 98, Issue 22, Pages 9229-9238Publisher
SPRINGER
DOI: 10.1007/s00253-014-6008-9
Keywords
E. coli; Recombinant protein; Inclusion bodies; Endotoxin; LPS; LPS-free
Categories
Funding
- INIA, (MINECO) Spain [RTA2012-00028-C02-02]
- Agencia de Gestio d'Ajuts Universitaris i de Recerca [2014SGR-132]
- Centro de Investigacion Biomedica en Red (CIBER) de Bioingenieria, Biomateriales y Nanomedicina - Instituto de Salud Carlos III
- MECD
- ICREA ACADEMIA Award
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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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