Title: Teaching an old pET new tricks: tuning of inclusion body formation and properties by a mixed feed system in E. coli
Language: English
Authors: Wurm, David Johannes 
Quehenberger, Julian  
Mildner, Julia 
Eggenreich, Britta 
Slouka, Christoph  
Schwaighofer, Andreas  
Wieland, Karin 
Lendl, Bernhard  
Rajamanickam, Vignesh 
Herwig, Christoph 
Spadiut, Oliver 
Category: Original Research Article
Issue Date: Jan-2018
Journal: Applied Microbiology and Biotechnology 
ISSN: 0175-7598
Abstract: 
Against the outdated belief that inclusion bodies (IBs) in Escherichia coli are only inactive aggregates of misfolded protein, and thus should be avoided during recombinant protein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.
Keywords: Escherichia coli BL21(DE3); pET expression system; Lactose; Inclusion body properties; Inclusion body size; Inclusion body purity
DOI: 10.1007/s00253-017-8641-6
Library ID: AC15321004
URN: urn:nbn:at:at-ubtuw:3-4884
Organisation: E166 - Institut für Verfahrenstechnik, Umwelttechnik und technische Biowissenschaften 
Publication Type: Article
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