Abstract
The key objectives for the optimisation of recombinant fermentation processes are the attainment of high yield together with the required product quality and the acceleration of process development. The overall strategy for process optimisation is designed to exploit the potential of the host cell by means of optimal control of the flux-ratios between biosynthesis of host cell proteins and recombinant proteins. To optimise the rate of recombinant protein expression, methods must be established (1) to monitor the metabolic load of the host cell resulting from recombinant protein overexpression, (2) to monitor and control plasmid copy number and (3) to adjust the expression rate to the metabolic load limits of the host cell. The metabolic load is derived from monitoring key molecules of the hierarchically organised cellular stress response regulatory networks and signal processing machinery. Tuning of recombinant protein expression is achieved by limited feed of inducer, based on the amount of biomass.
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References
Bayer K., Jungbauer A., Kramer W., Lettner H., Schönhofer W., Skias M., Steindl F., Taferner N. and Uhl K. Human rekombinant Superoxiddismutase. BioEngineering. 6 (1990): 24–30.
Breuer S., Marzban G., Cserjan-Puschmann M., Diirrschmid E. and Bayer K. Off-line quantitative monitoring of plasmid copy number in bacterial fermentation by capillary electrophoresis. Electrophoresis. 19 (1998): 2474–2478.
Cserjan-Puschmann M., Kramer W., Duerrschmid E., Striedner G. and Bayer K. Metabolic approaches for the optimisation of recombinant fermentation processes. Appl. Microbiol Biotechnol. 53–1 (1999): 43–50.
Grabherr R., Nilsson E. and Bayer K. Expression vectors with modified Co1E1 origin of replication for control of plasmid copy number, patent submitted.
Ferenci T. Adaptation to life at micromolar nutrient levels: the regulation of Escherichia coli glucose transport by endinduction and cAMP. FEMS Microbiol Rev. 18 (1996): 301–317.
Lengeier J.W., Drews G. and Schlegel H.G. Biology of the Prokaryotes. Georg Thieme Verlag Stuttgart. (1999): 491–522.
Porstmann T., Wietschke R., Schmechta H., Grunow R., Pergande M., Stachat S. and Baehr R. Rapid and sensitive enzyme immuno assay for Cu/Zn superoxide dismutase with polyclonal and monoclonal antibodies. Clin. Chim. Acta. 171 (1988): 1–10.
Studier F.W. and Moffat B.A. Use of the bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J. Mol Biol. 189(1986): 113–130.
Wróbel B. and Wegrzyn G. Replication regulation of Co1E1-like plasmids in amino acid-starved Escherichia coll Plasmid. 139 (1998): 48–62.
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© 2001 Springer Science+Business Media Dordrecht
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Striedner, G., Cserjan-Puschmann, M., Grabherr, R., Clementschitsch, F., Nilsson, E., Bayer, K. (2001). Metabolic Approaches for the Optimisation of Recombinant Fermentation Processes. In: Merten, OW., et al. Recombinant Protein Production with Prokaryotic and Eukaryotic Cells. A Comparative View on Host Physiology. Springer, Dordrecht. https://doi.org/10.1007/978-94-015-9749-4_15
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DOI: https://doi.org/10.1007/978-94-015-9749-4_15
Publisher Name: Springer, Dordrecht
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