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Reaction engineering studies for the production of 2-hydroxyisobutyric acid with recombinant Cupriavidus necator H 16

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Abstract

Recombinant Cupriavidus necator H 16 with a novel metabolic pathway using a cobalamin-dependent mutase was exploited to produce 2-hydroxyisobutyric acid (2-HIBA) from renewable resources through microbial fermentation. 2-HIBA production capacities of different strains of C. necator H 16 deficient in the PHB synthase gene and genetically engineered to enable the production of 2-HIBA from the intracellular PHB precursor (R)-3-hydroxybutyryl-CoA were evaluated in 48 parallel milliliter-scale stirred tank bioreactors (V = 11 mL). The effects of media composition, limitations, pH, and feed rate were studied with respect to the overall process performances of the different recombinant strains. 2-HIBA production was at a maximum at nitrogen limiting conditions and if the pH was controlled between 6.8 and 7.2 under fed-batch operating conditions (intermittent fructose addition). The final concentration of 2-HIBA was 7.4 g L−1 on a milliliter scale. Best reaction conditions identified on the milliliter scale were transferred to a laboratory-scale fed-batch process in a stirred tank bioreactor (V = 2 L). Two different process modes for the production of 2-HIBA, a single-phase and a dual-phase fermentation procedure, were evaluated and compared on a liter scale. The final concentration of 2-HIBA was 6.4 g L−1 on a liter scale after 2 days of cultivation.

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Acknowledgments

This research was financed and supported by Evonik Degussa GmbH and funded by the Bundesministerium für Ernährung, Landwirtschaft und Verbraucherschutz (BMELV). Consulting and assistance from A. Marx (Evonik Degussa GmbH), A. Thiessenhusen (Evonik Degussa GmbH), F. Erhardt (Evonik Degussa GmbH), and G. Faust are gratefully appreciated. Furthermore, this work was carried out under mentoring of the TUM Graduate School.

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Authors and Affiliations

  1. Lehrstuhl für Bioverfahrenstechnik, Technische Universität München, Boltzmannstr. 15, 85748, Garching, Germany

    Torben Hoefel & Dirk Weuster-Botz

  2. Evonik Degussa GmbH, Paul-Baumann-Str. 1, 45764, Marl, Germany

    Eva Wittmann & Liv Reinecke

Authors
  1. Torben Hoefel
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  2. Eva Wittmann
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  3. Liv Reinecke
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  4. Dirk Weuster-Botz
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Correspondence to Dirk Weuster-Botz.

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Hoefel, T., Wittmann, E., Reinecke, L. et al. Reaction engineering studies for the production of 2-hydroxyisobutyric acid with recombinant Cupriavidus necator H 16. Appl Microbiol Biotechnol 88, 477–484 (2010). https://doi.org/10.1007/s00253-010-2739-4

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  • DOI: https://doi.org/10.1007/s00253-010-2739-4

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