Minimizing inclusion body formation during recombinant protein production in Escherichia coli at bench and pilot plant scale

verfasst von

Frank Hoffmann, Joop Van Den Heuvel, Nadine Zidek, Ursula Rinas

Abstract

Many recombinant proteins partially aggregate into inclusion bodies during production in Escherichia coli in batch culture on defined medium. Production on complex medium, however, effectively prevented inclusion body formation of a β-galactosidase-HIVgp41 fusion protein for detection of anti-HIV antibodies which is produced at 42°C under control of a temperature- inducible expression system. Cells pre-conditioned by cultivation on complex medium before induction showed faster growth, higher product concentration and reduced inclusion body formation even when producing on defined medium. In contrast, for human basic fibroblast growth factor (hFGF-2) produced under control of the phage T7-promoter, medium composition could not reduce inclusion body formation even at 30°C. Here, slow production in high-cell density fed-batch mode using a defined medium with limited glucose feeding enabled the accumulation of 50mg product per gram cell dry mass exclusively in the soluble cell fraction, resulting in a volumetric concentration of more than 4g per litre hFGF-2. With the β-galactosidase fusion protein produced in fed-batch, over 100mg of product per gram cell dry mass accumulated in the soluble cell fraction. With a cell density of 100g cell dry mass per litre, this resulted in a volumetric concentration of 10g per litre of soluble β-galactosidase-HIVgp41 fusion protein. Thus, two approaches to balance heterologous protein production and host physiology are presented, which fit the needs of lab bench or pilot plant, respectively.

Externe Organisation(en)

Martin-Luther-Universität Halle-Wittenberg
Helmholtz-Zentrum für Infektionsforschung GmbH (HZI)

Typ

Artikel

Journal

Enzyme and microbial technology

Band

34

Seiten

235-241

Anzahl der Seiten

7

ISSN

0141-0229

Publikationsdatum

03.2004

Publikationsstatus

Veröffentlicht

Peer-reviewed

Ja

ASJC Scopus Sachgebiete

Biotechnologie, Bioengineering, Biochemie, Angewandte Mikrobiologie und Biotechnologie

Ziele für nachhaltige Entwicklung

SDG 3 – Gute Gesundheit und Wohlergehen

Elektronische Version(en)

https://doi.org/10.1016/j.enzmictec.2003.10.011 (Zugang: Geschlossen)