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Influence of the hydromechanical stress and temperature on growth and antibody fragment production with Bacillus megaterium

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Abstract

Bacillus megaterium was used for production of the lysozyme-specific recombinant scFv D1.3 antibody fragment. Key process parameters like the temperature and the hydromechanical stress play a very important role for significant product formation during process development or scale-up. In this study, the influence of these two variables on growth and recombinant antibody fragment production in a 2-L lab-scale bioreactor system was investigated using a central composite design. Especially a significant influence of the hydromechanical stress on antibody fragment production was detected in batch cultivations. While volumetric power inputs of about 0.5 kW/m3 (agitation rates around 500 min−1) are usually employed in batch cultivations, in this work maximal product concentration was found at a volumetric power input of about 0.06 kW/m3 (agitation rate around 250 min−1) and at a high cultivation temperature of 41 °C. The influence of the two process variables at single-cell level was estimated using flow cytometry too. The characterization was done by estimating the membrane potential giving a hint on bioprocess productivity and secretion capability: the best production was obtained through big cells with low specific membrane potential, which grew at low volumetric power inputs and high cultivation temperatures.

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Acknowledgments

This work was carried out as part of the DFG-Sonderforschungsbereich 578 and the BMBF-Nachwuchgruppe “Bioprozesstechnik”. The authors gratefully acknowledge financial support granted by the Deutsche Forschungsgemeinschaft (DFG) and the German Ministry of Research and Education (BMBF).

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The authors declare that they have no conflict of interest.

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  1. Eva Jordan

    Present address: Department of Biochemistry, Institute for Biochemistry and Biotechnology, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany

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  1. Institute of Biochemical Engineering, Technische Universität Braunschweig, Gausstrasse 17, 38106, Braunschweig, Germany

    Svenja Lüders, Florian David & Ezequiel Franco-Lara

  2. Department of Biotechnology, Institute for Biochemistry and Biotechnology, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany

    Miriam Steinwand, Eva Jordan, Michael Hust & Stefan Dübel

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  1. Svenja Lüders
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Correspondence to Ezequiel Franco-Lara.

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Lüders, S., David, F., Steinwand, M. et al. Influence of the hydromechanical stress and temperature on growth and antibody fragment production with Bacillus megaterium . Appl Microbiol Biotechnol 91, 81–90 (2011). https://doi.org/10.1007/s00253-011-3193-7

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