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Improved succinate production in Corynebacterium glutamicum by engineering glyoxylate pathway and succinate export system

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Abstract

A dual route for anaerobic succinate production was engineered into Corynebacterium glutamicum. The glyoxylate pathway was reconstructed by overexpressing isocitrate lyase, malate synthase and citrate synthase. The engineered strain produced succinate with a yield of 1.34 mol (mol glucose)−1. Further overexpression of succinate exporter, SucE, increased succinate yield to 1.43 mol (mol glucose)−1. Metabolic flux analysis revealed that the glyoxylate pathway was further activated by engineering succinate export system. Using an anaerobic fed-batch fermentation process, the final strain produced 926 mM succinate (= 109 g l−1) with an overall volumetric productivity of 9.4 mM h−1 and an average yield of 1.32 mol (mol glucose)−1.

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References

  • Beauprez JJ, De Mey M, Soetaert WK (2010) Microbial succinic acid production: natural versus metabolic engineered producers. Proc Biochem 45:1103–1114

    Article  CAS  Google Scholar 

  • Cok B, Tsiropoulos I, Roes AL, Patel MK (2013) Succinic acid production derived from carbohydrates: an energy and greenhouse gas assessment of a platform chemical toward a bio-based economy. Biofuel Bioprod Bioref. doi:10.1002/bbb.1427

    Google Scholar 

  • Eggeling L, Bott M (2005) Handbook of Corynebacterium glutamicum. CRC Press, Boca Raton

    Google Scholar 

  • Eikmanns BJ, Thum-Schmitz N, Eggeling L, Ludtke KU, Sahm H (1994) Nucleotide sequence, expression and transcriptional analysis of the Corynebacterium glutamicum gltA gene encoding citrate synthase. Microbiology 140:1817–1828

    Article  PubMed  CAS  Google Scholar 

  • Fukui K, Koseki C, Yamamoto Y, Nakamura J, Sasahara A, Yuji R, Hashiguchi K, Usuda Y, Matsui K, Kojima H, Abe K (2011) Identification of succinate exporter in Corynebacterium glutamicum and its physiological roles under anaerobic conditions. J Biotechnol 154:25–34

    Article  PubMed  CAS  Google Scholar 

  • Gerstmeir R, Cramer A, Dangel P, Schaffer S, Eikmanns BJ (2004) RamB, a novel transcriptional regulator of genes involved in acetate metabolism of Corynebacterium glutamicum. J Bacteriol 186:2798–2809

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  • Huhn S, Jolkver E, Krämer R, Marin K (2011) Identification of the membrane protein SucE and its role in succinate transport in Corynebacterium glutamicum. Appl Microbiol Biotechnol 89:327–335

    Article  PubMed  CAS  Google Scholar 

  • Kalinowski J, Bathe B, Bartels D, Bischoff N et al (2003) The complete Corynebacterium glutamicum ATCC 13032 genome sequence and its impact on the production of L-aspartate-derived amino acids and vitamins. J Biotechnol 104:5–25

    Article  PubMed  CAS  Google Scholar 

  • Kirchner O, Tauch A (2003) Tools for genetic engineering in the amino acid-producing bacterium Corynebacterium glutamicum. J Biotechnol 104:287–299

    Article  PubMed  CAS  Google Scholar 

  • Litsanov B, Brocker M, Bott M (2012) Towards homosuccinate fermentation: metabolic engineering of Corynebacterium glutamicum for anaerobic succinate production from glucose and formate. Appl Environ Microbiol 78:3325–3337

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  • Okino S, Noburyu R, Suda M, Jojima T, Inui M, Yukawa H (2008) An efficient succinic acid production process in a metabolically engineered Corynebacterium glutamicum strain. Appl Microbiol Biotechnol 81:459–464

    Article  PubMed  CAS  Google Scholar 

  • Reinscheid DJ, Eikmanns BJ, Sahm H (1994) Characterization of the isocitrate lyase gene from Corynebacterium glutamicum and biochemical analysis of the enzyme. J Bacteriol 176:3474–3483

    PubMed Central  PubMed  CAS  Google Scholar 

  • Sanchez AM, Bennett GN, San KY (2006) Batch culture characterization and metabolic flux analysis of succinate-producing Escherichia coli strains. Metab Eng 8:209–226

    Article  PubMed  CAS  Google Scholar 

  • Vemuri GN, Eiteman MA, Altman E (2002) Effects of growth mode and pyruvate carboxylase on succinic acid production by metabolically engineered strains of Escherichia coli. Appl Environ Microbiol 68:1715–1727

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  • Wang ZW, Chen T, Ma XH, Shen Z, Zhao XM (2011) Enhancement of riboflavin production with Bacillus subtilis by expression and site-directed mutagenesis of zwf and gnd gene from Corynebacterium glutamicum. Bioresour Technol 102:3934–3940

    Article  PubMed  CAS  Google Scholar 

  • Yukawa H, Omumasaba CA, Nonaka H, Kos P, Okai N, Suzuki N, Suda M, Tsuge Y, Watanabe J, Ikeda Y, Vertes AA, Inui M (2007) Comparative analysis of the Corynebacterium glutamicum group and complete genome sequence of strain R. Microbiology 153:1042–1058

    Article  PubMed  CAS  Google Scholar 

  • Zhu N, Xia H, Wang Z, Zhao X, Chen T (2013) Engineering of acetate recycling and citrate synthase to improve aerobic succinate production in Corynebacterium glutamicum. PLoS ONE 8:e60659

    Article  PubMed Central  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by National Program on Key Basic Research Project (2011CBA00804, 2012CB725203), National Natural Science Foundation of China (NSFC-21176182) and National High-tech R&D Program of China (2012AA02A702, 2012AA022103).

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Correspondence to Tao Chen.

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Zhu, N., Xia, H., Yang, J. et al. Improved succinate production in Corynebacterium glutamicum by engineering glyoxylate pathway and succinate export system. Biotechnol Lett 36, 553–560 (2014). https://doi.org/10.1007/s10529-013-1376-2

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  • DOI: https://doi.org/10.1007/s10529-013-1376-2

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