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Title
Teaching an old pET new tricks: tuning of inclusion body formation and properties by a mixed feed system in E. coli
AuthorWurm, David J. ; Quehenberger, Julian ; Mildner, Julia ; Eggenreich, Britta ; Slouka, Christoph ; Schwaighofer, Andrea ; Wieland, Karin ; Lendl, Bernhard ; Rajamanickam, Vignesh ; Herwig, Christoph ; Spadiut, Oliver
Published in
Applied Microbiology and Biotechnology, 2018, Vol. 102, Issue 2, page 667-676
PublishedSpringer Nature, 2018
LanguageEnglish
Document typeJournal Article
Keywords (EN)Escherichia coli BL21(DE3) / pET expression system / Lactose / Inclusion body properties / Inclusion body size / Inclusion body purity
ISSN0175-7598
URNurn:nbn:at:at-ubtuw:3-4884 Persistent Identifier (URN)
DOI10.1007/s00253-017-8641-6 
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Teaching an old pET new tricks: tuning of inclusion body formation and properties by a mixed feed system in E. coli [2.8 mb]
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Abstract (English)

Against the outdated belief that inclusion bodies (IBs) in Escherichia coli are only inactive aggregates of misfolded protein, and thus should be avoided during recombinant protein production, numerous biopharmaceutically important proteins are currently produced as IBs. To obtain correctly folded, soluble product, IBs have to be processed, namely, harvested, solubilized, and refolded. Several years ago, it was discovered that, depending on cultivation conditions and protein properties, IBs contain partially correctly folded protein structures, which makes IB processing more efficient. Here, we present a method of tailored induction of recombinant protein production in E. coli by a mixed feed system using glucose and lactose and its impact on IB formation. Our method allows tuning of IB amount, IB size, size distribution, and purity, which does not only facilitate IB processing, but is also crucial for potential direct applications of IBs as nanomaterials and biomaterials in regenerative medicine.

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CC-BY-License (4.0)Creative Commons Attribution 4.0 International License