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An efficient succinic acid production process in a metabolically engineered Corynebacterium glutamicum strain

Applied Microbiology and Biotechnology volume 81, pages 459–464 (2008)Cite this article

Abstract

A Corynebacterium glutamicum strain (ΔldhA-pCRA717) that overexpresses the pyc gene encoding pyruvate carboxylase while simultaneously exhibiting a disrupted ldhA gene encoding l-lactate dehydrogenase was investigated in detail for succinic acid production. Succinic acid was shown to be efficiently produced at high-cell density under oxygen deprivation with intermittent addition of sodium bicarbonate and glucose. Succinic acid concentration reached 1.24 M (146 g l−1) within 46 h. The yields of succinic acid and acetic acid from glucose were 1.40 mol mol−1 (0.92 g g−1) and 0.29 mol mol−1 (0.10 g g−1), respectively. The succinic acid production rate and yield depended on medium bicarbonate concentration rather than glucose concentration. Consumption of bicarbonate accompanied with succinic acid production implied that added bicarbonate was used for succinic acid synthesis.

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References

  • Chattrjee R, Millard CS, Champion K, Clark DP, Donnelly MI (2001) Mutation of the ptsG gene results in increased production of succinate in fermentation of glucose by Escherichia coli. Appl Environ Microbiol 67:148–154

    Article  Google Scholar 

  • Donnelly MI, Millard CS, Stols L (1998) Mutant E. coli strain with increased succinic acid production. United States Patent 5,770,435

  • Glassner DA, Datta R (1992) Process for the production and purification of succinic acid. United States Patent 5,143,834

  • Guettler MV, Jain MK, Rumler D (1996) Method for making succinic acid, bacterial variants for use in the process, and methods for obtaining variants. United States Patent 5,573,931

  • Gutknecht J, Bisson MA, Tosteson FC (1977) Diffusion of carbon dioxide through lipid bilayer membranes: effects of carbonic anhydrase, bicarbonate, and unstirred layers. J Gen Physiol 69:779–794

    CAS  Article  Google Scholar 

  • Ikeda M, Nakagawa S (2003) The Corynebacterium glutamicum genome: features and impacts on biotechnological processes. Appl Microbiol Biotechnol 62:99–109

    CAS  Article  Google Scholar 

  • Inui M, Murakami S, Okino S, Kawaguchi H, Vertès AA, Yukawa H (2004) Metabolic analysis of Corynebacterium glutamicum during lactate and succinate productions under oxygen deprivation conditions. J Mol Microbiol Biotechnol 7:182–196

    CAS  Article  Google Scholar 

  • Kalinowski J, Bathe B, Bartels D, Bischoff N, Bott M, Burkovski A, Dusch N, Eggeling L, Eikmanns BJ, Gaigalat L, Goesmann A, Hartmann M, Huthmacher K, Kramer R, Linke B, McHardy AC, Meyer F, Mockel B, Pfefferle W, Puhler A, Rey DA, Ruckert C, Rupp O, Sahm H, Wendisch VF, Wiegrabe I, Tauch A (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

    CAS  Article  Google Scholar 

  • Kawaguchi H, Vertès AA, Okino S, Inui M, Yukawa H (2006) Engineering of a xylose metabolic pathway in Corynebacterium glutamicum. Appl Environ Microbiol 72:3418–3428

    CAS  Article  Google Scholar 

  • Kawaguchi H, Sasaki M, Vertès AA, Inui M, Yukawa H (2008) Engineering of an L-arabinose metabolic pathway in Corynebacterium glutamicum. Appl Microbiol Biotechnol 77:1053–1062

    CAS  Article  Google Scholar 

  • Lee PC, Lee WG, Kwon S, Lee SY, Chang HN (2000) Batch and continuous cultivation of Anaerobiospirillum succiniciproducens for the production of succinic acid from whey. Appl Microbiol Biotechnol 54:23–27

    CAS  Article  Google Scholar 

  • Lee PC, Lee SY, Hong SH, Chang HN, Park SC (2003) Biological conversion of wood hydrolysate to succinic acid by Anaerobiospirillum succiniciproducens. Biotechnol Lett 25:111–114

    CAS  Article  Google Scholar 

  • McKinlay JB, Vieille C, Zeikus JG (2007) Prospects for a bio-based succinate industry. Appl Microbiol Biotechnol 76:727–740

    CAS  Article  Google Scholar 

  • Merlin C, Masters M, McAteer S, Coulson A (2003) Why is carbonic anhydrase essential to Escherichia coli? J Bacteriol 185:6415–6424

    CAS  Article  Google Scholar 

  • Meynial-Salles I, Dorotyn S, Soucaille P (2008) A new process for the continuous production of succinic acid from glucose at high yield, titer, and productivity. Biotechnol Bioeng 99:129–135

    CAS  Article  Google Scholar 

  • Okino S, Inui M, Yukawa H (2005) Production of organic acids by Corynebacterium glutamicum under oxygen deprivation. Appl Microbiol Biotechnol 68:475–480

    CAS  Article  Google Scholar 

  • Samuelov NS, Datta R, Jain MK, Zeikus JG (1999) Whey fermentation by Anaerobiospirillum succiniciproducens for production of a succinate-based animal feed additive. Appl Environ Microbiol 65:2260–2263

    CAS  Article  Google Scholar 

  • Sahm H, Eggeling L, de Graaf AA (2000) Pathway analysis and metabolic engineering in Corynebacterium glutamicum. Biol. Chem. 381:899–910

    CAS  Article  Google Scholar 

  • Sauer M, Porro D, Mattanovich D, Branduardi P (2008) Microbial production of organic acids: expanding the markets. Trends Biotechnol 26:100–108

    CAS  Article  Google Scholar 

  • Schreiner ME, Fiur D, Holatko J, Patek M, Eikmanns BJ (2005) E1 enzyme of the pyruvate dehydrogenase complex in Corynebacterium glutamicum: molecular analysis of the gene and phylogenetic aspects. J Bacteriol 187:6005–6018

    CAS  Article  Google Scholar 

  • Vemuri GN, Eiteman MA, Altman E (2002) Succinate production in dual-phase Escherichia coli fermentations depends on the time of transition from aerobic to anaerobic conditions. J Ind Microbiol Biotechnol 28:325–332

    CAS  Article  Google Scholar 

  • Vertès AA, Inui M, Yukawa H (2005) Manipulating corynebacteria, from individual genes to chromosomes. Appl Environ Microbiol 71:7633–7642

    Article  Google Scholar 

  • Werpy T, Petersen G (2004) Top Value-Added Chemicals from Biomass, Volume I: Results of Screening for Potential Candidates from Sugars and Synthesis Gas, http://www.eere.energy.gov/biomass/pdfs/35523.pdf. Cited August 2004

  • Wu H, Li ZM, Zhou L, Ye Q (2007) Improved succinic acid production in the anaerobic culture of an Escherichia coli pflB ldhA double mutant as a result of enhanced anaplerotic activities in the preceding aerobic culture. Appl Environ Microbiol 73:7837–7843

    CAS  Article  Google Scholar 

  • Yasuda K, Jojima T, Suda M, Okino S, Inui M, Yukawa H (2007) Analyses of the acetate-producing pathways in Corynebacterium glutamicum under oxygen-deprived conditions. Appl Microbiol Biotechnol 77:853–860

    CAS  Article  Google Scholar 

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

    CAS  Article  Google Scholar 

  • Zeikus JG, Jain MK, Elankovan P (1999) Biotechnology of succinic acid production and markets for derived industrial products. Appl Microbiol Biotechnol 51:542–552

    Article  Google Scholar 

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Affiliations

  1. Research Institute of Innovative Technology for the Earth, 9-2 Kizugawadai, Kizugawa, Kyoto, 619-0292, Japan

    Shohei Okino, Ryoji Noburyu, Masako Suda, Toru Jojima, Masayuki Inui & Hideaki Yukawa

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  1. Shohei Okino
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  2. Ryoji Noburyu
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  3. Masako Suda
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  4. Toru Jojima
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  5. Masayuki Inui
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  6. Hideaki Yukawa
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Correspondence to Hideaki Yukawa.

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Okino, S., Noburyu, R., Suda, M. et al. An efficient succinic acid production process in a metabolically engineered Corynebacterium glutamicum strain. Appl Microbiol Biotechnol 81, 459–464 (2008). https://doi.org/10.1007/s00253-008-1668-y

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  • DOI: https://doi.org/10.1007/s00253-008-1668-y

Keywords

  • Succinic acid
  • Corynebacterium glutamicum
  • Pyruvate carboxylase