Teaching an old pET new tricks: tuning of inclusion body formation and properties by a mixed feed system in E. coli

Wurm, David Johannes; Quehenberger, Julian; Mildner, Julia; Eggenreich, Britta; Slouka, Christoph; Schwaighofer, Andreas; Wieland, Karin; Lendl, Bernhard; Rajamanickam, Vignesh; Herwig, Christoph; Spadiut, Oliver

doi:10.1007/s00253-017-8641-6

Record link:

https://resolver.obvsg.at/urn:nbn:at:at-ubtuw:3-4884
http://hdl.handle.net/20.500.12708/91

Title:

Teaching an old pET new tricks: tuning of inclusion body formation and properties by a mixed feed system in E. coli

Citation:

Wurm, D. J., Quehenberger, J., Mildner, J., Eggenreich, B., Slouka, C., Schwaighofer, A., Wieland, K., Lendl, B., Rajamanickam, V., Herwig, C., & Spadiut, O. (2018). Teaching an old pET new tricks: tuning of inclusion body formation and properties by a mixed feed system in E. coli. Applied Microbiology and Biotechnology, 102(2), 667–676. https://doi.org/10.1007/s00253-017-8641-6

Publisher DOI:

10.1007/s00253-017-8641-6

CatalogPlus:

AC15321004

Publication Type:

Article - Original Research Article

Language:

English

Organisational Unit:

E166 - Institut für Verfahrenstechnik, Umwelttechnik und technische Biowissenschaften

Journal:

Applied Microbiology and Biotechnology

ISSN:

0175-7598

Date (published):

Jan-2018

Number of Pages:

Publisher:

SPRINGER

Peer reviewed:

Yes

Keywords:

Escherichia coli BL21(DE3); pET expression system; Lactose; Inclusion body properties; Inclusion body size; Inclusion body purity

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Against the outdated belief that inclusion bodies (IBs) in Escherichia coli are only inactive aggregates of misfolded protein, and thus should be avoided during recombinant protein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.

License:

CC BY 4.0