Production of functional inclusion bodies in endotoxin-free Escherichia coli

Fabián Rueda, Olivia Cano-Garrido, Uwe Mamat, Kathleen Wilke, Joaquin Seras-Franzoso, Elena García-Fruitós, Antonio Villaverde

Research output: Contribution to journalArticleResearchpeer-review

33 Citations (Scopus)

Abstract

© 2014, Springer-Verlag Berlin Heidelberg. 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.
Original languageEnglish
Pages (from-to)9229-9238
JournalApplied Microbiology and Biotechnology
Volume98
DOIs
Publication statusPublished - 1 Jan 2014

Keywords

  • E. coli
  • Endotoxin
  • Inclusion bodies
  • LPS
  • LPS-free
  • Recombinant protein

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