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Assessment of robustness against dissolved oxygen/substrate oscillations for C. glutamicum DM1933 in two-compartment bioreactor

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Abstract

Corynebacterium glutamicum is an important organism for industrial biotechnology; particularly, in amino acid production (e.g. l-lysine). Production scales often reach reactor working volumes of several hundred cubic meters, which triggers inhomogeneous distribution of substrates and dissolved gasses due to increasing mixing times. Individual cells which follow the flow profile through the reactor are experiencing oscillating microenvironments. Oscillations can have an influence on the process performance, which is a subject of scale-down experiments. In this work, l-lysine-producing C. glutamicum DM1933 was assessed for its robustness against continuous dissolved oxygen and substrate supply oscillation in two-compartment scale-down bioreactors. Aerobic, substrate-limited stirred tank and non-aerated, substrate-excess plug flow compartments were applied for oscillation. Inhomogeneity of substrate and oxygen supply was observed to cause rapid side product turnover, redistribution of oxygen uptake from oxygen limited into fully aerobic zones, and intermediate medium acidification. However, process inhomogeneity did not impair productivity or growth at plug flow residence times of several minutes. In a focused analysis of proteome, metabolome, transcriptome, and other physiological parameters, no changes were identified in response to process inhomogeneity. In conclusion, fed-batch processes with C. glutamicum DM1933 possess remarkable robustness against oxygen and substrate supply oscillation, which is a unique property in the field of published scale-down studies. Microbial physiology of C. glutamicum appears to be ideally adapted to both homogeneous and inhomogeneous conditions. This ensures exceptional suitability for cultivation at increased mixing times, which is suggested to constitute an important basis for the long-lasting success in large scale bioprocess application.

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Acknowledgments

The authors thank the Bundesministerium für Bildung und Forschung (BMBF) for funding in the cluster project “Corynebacterium: Improving flexibility and fitness for industrial production” (Grant No. 0315589A), and the fruitful cooperation with industrial project partner Evonik Industries. The authors also thank Nicole Paczia, Max von Haugwitz, Petra Geilenkirchen, and Matthias Moch for guidance and conduction of metabolome analysis procedures, and express special thanks to Raphael Voges for the conduction of targeted proteome analysis by a recently established protocol.

Conflict of interest

The authors have declared that no competing interest exists.

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Authors and Affiliations

  1. Institute of Bio- and Geosciences, IBG-1: Biotechnology, Systems Biotechnology, Forschungszentrum Jülich GmbH, 52425, Jülich, Germany

    Friedrich Käß, Ioanna Hariskos, Andrea Michel, Hans-Jürgen Brandt, Wolfgang Wiechert & Marco Oldiges

  2. Chair of Bioprocess Engineering, Department of Biotechnology, Technische Universität Berlin, Ackerstrasse 71-76, 13355, Berlin, Germany

    Robert Spann, Stefan Junne & Peter Neubauer

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  1. Friedrich Käß
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  2. Ioanna Hariskos
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Correspondence to Marco Oldiges.

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Käß, F., Hariskos, I., Michel, A. et al. Assessment of robustness against dissolved oxygen/substrate oscillations for C. glutamicum DM1933 in two-compartment bioreactor. Bioprocess Biosyst Eng 37, 1151–1162 (2014). https://doi.org/10.1007/s00449-013-1086-0

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