High cell density fermentation of recombinant Escherichia coli expressing human interferon alpha 1 |
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Authors: | Dieter Riesenberg Klaus Menzel Volkmar Schulz Klaus Schumann Gerhard Veith Georg Zuber Wolfgang A Knorre |
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Institution: | (1) Zentralinstitut für Mikrobiologie und experimentelle Therapie der AdW der DDR, Beutenbergstrasse 11, 6900 Jena, German Democratic Republic |
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Abstract: | Summary A defined medium was developed which, by means of a specific fed-batch mode, allows growth of the recombinant Escherichia coli strain TG1 (pBB210) up to a cell density of 60 g dry weight/l. Apart from glucose and aqueous ammonia fed as carbon and nitrogen sources, it was unnecessary to supply other nutrients or O2-enriched air. Aqueous ammonia also served for pH control. The pO2 level was kept at 20% saturation via closed-loop controls operating the two output variables of stirrer speed and glucose feeding rate. This fed-batch method prevented significant accumulation of acetate and other metabolic by-products. The recombinant E. coli expressed interferon alpha 1 more efficiently at a lower specific growth rate ( Pr 0.15 h–1) than at the maximum specific growth rate ( max = 0.45 h–1). Therefore, fermentation in the batch phase at max was only allowed to continue up to a medium cell density. In the succeeding fed-batch phase, the specific growth rate was reduced to Pr by increasing the stirrer speed according to an empirically developed time scale.
Offprint requests to: D. Riesenberg |
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