Teaching an old pET new tricks: tuning of inclusion body formation and properties by a mixed feed system in E. coli
| Autor: | Julia Mildner, David J. Wurm, Bernhard Lendl, Christoph Herwig, Andreas Schwaighofer, Karin Wieland, Britta Eggenreich, Christoph Slouka, Oliver Spadiut, Julian Quehenberger, Vignesh Rajamanickam |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Protein Folding Inclusion body size Lactose medicine.disease_cause Applied Microbiology and Biotechnology Inclusion bodies law.invention 03 medical and health sciences chemistry.chemical_compound Inclusion body properties Protein structure law Escherichia coli medicine Inclusion body purity Inclusion Bodies Escherichia coli Proteins Mixed feed pET expression system General Medicine Biotechnological Products and Process Engineering Recombinant Proteins Culture Media Glucose 030104 developmental biology Solubility Biochemistry chemistry Recombinant protein production Recombinant DNA Protein folding Escherichia coli BL21(DE3) Biotechnology |
| Zdroj: | Applied Microbiology and Biotechnology |
| ISSN: | 1432-0614 0175-7598 |
| DOI: | 10.1007/s00253-017-8641-6 |
| Popis: | Against the outdated belief that inclusion bodies (IBs) in Escherichia coli are only inactive aggregates of misfolded protein, and thus should be avoided during recombinant protein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. Electronic supplementary material The online version of this article (10.1007/s00253-017-8641-6) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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