Segregation to non-dividing cells in recombinant Escherichia coli fed-batch fermentation processes |
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| Authors: | Heléne Sundström Fredrik Wållberg Erika Ledung Bo Norrman Christopher J Hewitt and Sven-Olof Enfors |
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| Institution: | (1) Department of Biochemical Engineering, University College London, Torrington Place, London, WC1E 7JE, UK;(2) Department of Chemical and Biochemical Engineering, Center for BioProcess Engineering, Technical University of Denmark, 2800 Lyngby, Denmark;(3) Bioprocess R and D, Merck Research Laboratories, Rahway, NJ 07062-0900, USA;(4) Department of Chemical Engineering, Interdisciplinary Centre for Biological Engineering, Loughborough University, Leicestershire, LE11 3TU, UK; |
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| Abstract: | In Escherichia coli fermentation processes, a drastic drop in viable cell count as measured by the number of colony forming units per ml (c.f.u. ml–1) is often observed. This phenomenon was investigated in a process for the production of the recombinant fusion protein, promegapoietin (PMP). After induction, the number of c.f.u. ml–1 dropped to  10% of its maximum though the biomass concentration continued to increase. Flow cytometric analysis of viability and intracellular concentration of PMP showed that almost all cells were alive and contributed to the production. Thus, the drop in the number of c.f.u. ml–1 probably reflects a loss of cell division capability rather than cell death. |
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| Keywords: | bacterial cell division Escherichia coli fed-batch fermentation flow cytometry viability |
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