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High yield production of four-carbon dicarboxylic acids by metabolically engineered Escherichia coli

  • Metabolic Engineering and Synthetic Biology - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Several metabolic engineered Escherichia coli strains were constructed and evaluated for four-carbon dicarboxylic acid production. Fumarase A, fumarase B and fumarase C single, double and triple mutants were constructed in a ldhA adhE mutant background overexpressing the pyruvate carboxylase from Lactococcus lactis. All the mutants produced succinate as the main four-carbon (C4) dicarboxylic acid product when glucose was used as carbon source with the exception of the fumAC and the triple fumB fumAC deletion strains, where malate was the main C4-product with a yield of 0.61–0.67 mol (mole glucose)−1. Additionally, a mdh mutant strain and a previously engineered high-succinate-producing strain (SBS550MG-Cms pHL413-Km) were investigated for aerobic malate production from succinate. These strains produced 40.38 mM (5.41 g/L) and 50.34 mM (6.75 g/L) malate with a molar yield of 0.53 and 0.55 mol (mole succinate)−1, respectively. Finally, by exploiting the high-succinate production capability, the strain SBS550MG-Cms243 pHL413-Km showed significant malate production in a two-stage process from glucose. This strain produced 133 mM (17.83 g/L) malate in 47 h, with a high yield of 1.3 mol (mole glucose)−1 and productivity of 0.38 g L−1 h−1.

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Acknowledgements

The authors acknowledge funding from grants from the National Institutes of Health (NIH GM090152) and NSF (CBET-0828516). The authors would like to thank Auritra Mallick for performing preliminary experiments.

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Authors and Affiliations

  1. Department of Bioengineering, Rice University, 6500 Main Street, Houston, TX, 77030, USA

    Irene Martinez, Haijun Gao & Ka-Yiu San

  2. Escuela de Ingeniería Bioquímica, Pontificia Universidad Católica de Valparaíso, Valparaiso, Chile

    Irene Martinez

  3. Department of BioSciences, Rice University, Houston, TX, USA

    George N. Bennett

  4. Department of Chemical and Biomolecular Engineering, Rice University, Houston, TX, USA

    George N. Bennett & Ka-Yiu San

Authors
  1. Irene Martinez
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  2. Haijun Gao
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  3. George N. Bennett
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  4. Ka-Yiu San
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Correspondence to Ka-Yiu San.

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Martinez, I., Gao, H., Bennett, G.N. et al. High yield production of four-carbon dicarboxylic acids by metabolically engineered Escherichia coli . J Ind Microbiol Biotechnol 45, 53–60 (2018). https://doi.org/10.1007/s10295-017-1991-3

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  • DOI: https://doi.org/10.1007/s10295-017-1991-3

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