direkt zum Inhalt springen

direkt zum Hauptnavigationsmenü

Sie sind hier

TU Berlin

Inhalt des Dokuments

Es gibt keine deutsche Übersetzung dieser Webseite.


Suche nach Publikationen

Limiting factors in Escherichia coli fed-batch production of recombinant proteins
Zitatschlüssel Sanden.2003
Autor Sandén, Anna Maria and Prytz, Ingela and Tubulekas, Ioannis and Förberg, Cecilia and Le, Ha and Hektor, Andrea and Neubauer, Peter and Pragai, Zoltan and Harwood, Colin and Ward, Alan and Picon, Antonia and de Mattos, Joost Teixeira and Postma, Pieter and Farewell, Anne and Nyström, Thomas and Reeh, Solvejg and Pedersen, Steen and Larsson, Gen
Seiten 158–166
Jahr 2003
ISSN 1097-0290
DOI 10.1002/bit.10457
Journal Biotechnology and bioengineering
Jahrgang 81
Nummer 2
Zusammenfassung Fed-batch production of recombinant beta-galactosidase in E. coli was studied with respect to the specific growth rate at induction. The cultivations were designed to induce protein production by IPTG at a glucose feed rate corresponding to high mu = 0.5 h(-1)) or low (mu = 0.1 h(-1)) specific growth rate. Protein production rate was approximately 100% higher at the higher specific growth rate, resulting in the accumulation of beta-galactosidase up to 30% of the total cell protein. Transcription analysis showed that beta-galactosidase-specific messenger RNA was immediately formed after induction (<5 min), but the amount was the same in both cases and was thus not the initial limiting factor. The content of ribosomes, as represented by rRNA, rapidly decreased with specific growth rate from a relative level of 100%, at the high specific growth rate, to 20% at the low specific growth rate. At high specific growth rate, ribosomes were additionally degraded upon induction due to the high production level. Translation therefore seemed to be the initial limiting factor of the protein synthesis capacity. The alarmone guanosine tetraphosphate increased at both high and low feed level inductions, indicating an induction-forced starvation of charged tRNA and/or glucose. The altered physiological status was also detected by the formation of acetic acid. However, the higher production rate resulted in high-level accumulation of acetic acid, which was absent at low feed rate production. Acetic acid production is thus coupled to the high product formation rate and is proposed to be due either to a precursor drain of Krebs cycle intermediates and a time lag before induction of the glyoxalate shunt, or to single amino acid overflow, since the model product is relatively poor in glycin and alanin. In conclusion, it is proposed that production at high specific growth rate becomes precursor-limited, while production at low specific growth rate is carbon- and/or energy-limited.
Download Bibtex Eintrag

Zusatzinformationen / Extras


Schnellnavigation zur Seite über Nummerneingabe