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Comparative study on central metabolic fluxes of Bacillus megaterium strains in continuous culture using 13C labelled substrates

Abstract

Fluxes of central carbon metabolism [glycolysis, pentose phosphate pathway (PPP), tricarboxylic acid cycle (TCA cycle), biomass formation] were determined for several Bacillus megaterium strains (DSM319, WH320, WH323, MS941) in C- and N-limited chemostat cultures by 13C labelling experiments. The labelling patterns of proteinogenic amino acids were analysed by GC/MS and therefrom flux ratios at important nodes within the metabolic network could be calculated. On the basis of a stoichiometric metabolic model flux distributions were estimated for the different B. megaterium strains used at various cultivation conditions. Generally all strains exhibited similar metabolic flux distributions, however, several significant changes were found in (1) the glucose flux entering the PPP via the oxidative branch, (2) the reversibilities within the PPP, (3) the relative fluxes of pyruvate and acetyl-CoA fed to the TCA cycle, (4) the fluxes around the pyruvate node involving a futile cycle.

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Acknowledgments

This work was carried out as part of the DFG-Sonderforschungsbereich 578 and the authors gratefully acknowledge financial support granted by the Deutsche Forschungsgemeinschaft. Thanks are also due to the Structure Biology Department of the Helmholtz Zentrum für Infektionsforschung (HZI), especially to Dr Manfred Nimtz and Claudia Hanko for their support in GC/MS analysis.

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Correspondence to Tobias Fürch.

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Fürch, T., Hollmann, R., Wittmann, C. et al. Comparative study on central metabolic fluxes of Bacillus megaterium strains in continuous culture using 13C labelled substrates. Bioprocess Biosyst Eng 30, 47–59 (2007). https://doi.org/10.1007/s00449-006-0095-7

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Keywords

  • Bacillus megaterium
  • Metabolic flux analysis
  • Central carbon pathways
  • Futile cycle