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Glucose consumption rate critically depends on redox state in Corynebacterium glutamicum under oxygen deprivation

  • Applied genetics and molecular biotechnology
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Abstract

Rapid sugar consumption is important for the microbial production of chemicals and fuels. Here, we show that overexpression of the NADH dehydrogenase gene (ndh) increased glucose consumption rate in Corynebacterium glutamicum under oxygen-deprived conditions through investigating the relationship between the glucose consumption rate and intracellular NADH/NAD+ ratio in various mutant strains. The NADH/NAD+ ratio was strongly repressed under oxygen deprivation when glucose consumption was accelerated by the addition of pyruvate or sodium hydrogen carbonate. Overexpression of the ndh gene in the wild-type strain under oxygen deprivation decreased the NADH/NAD+ ratio from 0.32 to 0.13, whereas the glucose consumption rate increased by 27 %. Similarly, in phosphoenolpyruvate carboxylase gene (ppc)- or malate dehydrogenase gene (mdh)-deficient strains, overexpression of the ndh gene decreased the NADH/NAD+ ratio from 1.66 to 0.37 and 2.20 to 0.57, respectively, whereas the glucose consumption rate increased by 57 and 330 %, respectively. However, in a lactate dehydrogenase gene (L-ldhA)-deficient strain, although the NADH/NAD+ ratio decreased from 5.62 to 1.13, the glucose consumption rate was not markedly altered. In a tailored d-lactate-producing strain, which lacked ppc and L-ldhA genes, but expressed D-ldhA from Lactobacillus delbrueckii, overexpression of the ndh gene decreased the NADH/NAD+ ratio from 1.77 to 0.56, and increased the glucose consumption rate by 50 %. Overall, the glucose consumption rate was found to be inversely proportional to the NADH/NAD+ ratio in C. glutamicum cultured under oxygen deprivation. These findings could provide an option to increase the productivity of chemicals and fuels under oxygen deprivation.

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References

  • Bartek T, Blombach B, Lang S, Eikmanns BJ, Wiechert W, Oldiges M, Nöh K, Noack S (2011) Comparative 13C metabolic flux analysis of pyruvate dehydrogenase complex-deficient, l-valine-producing Corynebacterium glutamicum. Appl Environ Microbiol 77:6644–6652

  • Blombach B, Riester T, Wieschalka S, Ziert C, Youn JW, Wendisch VF, Eikmanns BJ (2011) Corynebacterium glutamicum tailored for efficient isobutanol production. Appl Environ Microbiol 77:3300–3310

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Danshina PV, Schmalhausen EV, Avetisyan AV, Muronetz VI (2001) Mildly oxidized glyceraldehyde-3-phosphate dehydroge-nase as a possible regulator of glycolysis. IUBMB Life 51:309–314

    Article  CAS  PubMed  Google Scholar 

  • Dominguez H, Rollin C, Guyonvarch A, Guerquin-Kern JL, Cocaign-Bousquet M, Lindley ND (1998) Carbon-flux distribution in the central metabolic pathways of Corynebacterium glutamicum during growth on fructose. Eur J Biochem 254:96–102

    Article  CAS  PubMed  Google Scholar 

  • Duggleby RG, Dennis DT (1974) Nicotin-amide adenine dinucleotide-specific glyceral-dehyde 3-phosphate dehydrogenase from Pisum sativum. Assay and steady state kinetics. J Biol Chem 249:167–174

  • Garrigues C, Loubiere P, Lindley ND, Cocaign-Bousquet M (1997) Control of the shift from homolactic acid to mixed-acid fermentation in Lactococcus lactis: predominant role of the NADH/NAD+ ratio. J Bacteriol 179:5282–5287

    CAS  PubMed Central  PubMed  Google Scholar 

  • Hasegawa S, Uematsu K, Natsuma Y, Suda M, Hiraga K, Jojima T, Inui M, Yukawa H (2012) Improvement of the redox balance increases l-valine production by Corynebacterium glutamicum under oxygen deprivation conditions. Appl Environ Microbiol 78:865–875

  • Hasegawa S, Suda M, Uematsu K, Natsuma Y, Hiraga K, Jojima T, Inui M, Yukawa H (2013) Engineering of Corynebacterium glutamicum for high-yield l-valine production under oxygen deprivation conditions. Appl Environ Microbiol 79:1250–1257

  • Heux S, Cachon R, Dequin S (2006) Cofactor engineering in Saccharomyces cerevisiae: Expression of a H2O-forming NADH oxidase and impact on redox metabolism. Metab Eng 8:303–314

    Article  CAS  PubMed  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

    Article  CAS  PubMed  Google Scholar 

  • Inui M, Suda M, Okino S, Nonaka H, Puskas LG, Vertès AA, Yukawa H (2007) Transcriptional profiling of Corynebacterium glutamicum metabolism during organic acid production under oxygen deprivation conditions. Microbiology 153:2491–2504

    Article  CAS  PubMed  Google Scholar 

  • Jaworowski A, Mayo G, Shaw DC, Campbell HD, Young IG (1981) Characterization of the respiratory NADH dehydrogenase of Escherichia coli and reconstitution of NADH oxidase in ndh mutant membrane vesicles. Biochemistry 20:3621–3628

    Article  CAS  PubMed  Google Scholar 

  • Kinoshita S (1985) Glutamic acid bacteria. In: Demain AL, Solomon NA (eds) Biology of industrial microorganisms. Benjamin Cummings, London, pp 115–146

    Google Scholar 

  • Koebmann BJ, Solem C, Pedersen MB, Nilsson D, Jensen PR (2002) Expression of genes encoding F1-ATPase results in uncoupling of glycolysis from biomass production in Lactococcus lactis. Appl Environ Microbiol 68:4274–4282

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Leif H, Sled VD, Ohnishi T, Weiss H, Friedrich T (1995) Isolation and characterization of the proton-translocating NADH: ubiquinone oxidoreductase from Escherichia coli. Eur J Biochem 230:538–548

    Article  CAS  PubMed  Google Scholar 

  • Liang L, Liu R, Wang G, Gou D, Ma J, Chen K, Jiang M, Wei P, Ouyang P (2012) Regulation of NAD(H) pool and NADH/NAD(+) ratio by overexpression of nicotinic acid phosphoribosyltransferase for succinic acid production in Escherichia coli NZN111. Enzyme Microb Technol 51:286–293

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Lopez de Felipe F, Kleerebezem M, de Vos WM, Hugenholtz J (1998) Cofactor engineering: a novel approach to metabolic engineering in Lactococcus lactis by controlled expression of NADH oxidase. J Bacteriol 180:3804–3808

    CAS  PubMed Central  PubMed  Google Scholar 

  • Matsushita K, Ohnishi T, Kaback HR (1987) NADH-ubiquinone oxidoreductases of the Escherichia coli aerobic respiratory chain. Biochemistry 26:7732–7737

    Article  CAS  PubMed  Google Scholar 

  • Matsushita K, Otofuji A, Iwahashi M, Toyama H, Adachi O (2001) NADH dehydrogenase of Corynebacterium glutamicum. Purification of an NADH dehydrogenase II homolog able to oxidize NADPH. FEMS Microbiol Lett 204:271–276

    Article  CAS  PubMed  Google Scholar 

  • Molenaar D, van der Rest ME, Drysch A, Yucel R (2000) Functions of the membrane-associated and cytoplasmic malate dehydrogenases in the citric acid cycle of Corynebacterium glutamicum. J Bacteriol 182:6884–6891

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Nakayama K, Kitada S, Kinoshita S (1961) Studies on lysine fermentation I. The control mechanism on lysine accumulation by homoserine and threonine. J Gen Appl Microbiol 7:145–154

    Article  CAS  Google Scholar 

  • Nantapong N, Kugimiya Y, Toyama H, Adachi O, Matsushita K (2004) Effect of NADH dehydrogenase-disruption and over-expression on respiration-related metabolism in Corynebacterium glutamicum KY9714. Appl Microbiol Biotechnol 66:187–193

    Article  CAS  PubMed  Google Scholar 

  • Nantapong N, Otofuji A, Migita CT, Adachi O, Toyama H, Matsushita K (2005) Electron transfer ability from NADH to menaquinone and from NADPH to oxygen of type II NADH dehydrogenase of Corynebacterium glutamicum. Biosci Biotechnol Biochem 69:149–159

    Article  CAS  PubMed  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

    Article  CAS  PubMed  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  CAS  PubMed  Google Scholar 

  • Palmfeldt J, Paese M, Hahn-Hagerdal B, Van Niel EW (2004) The pool of ADP and ATP regulates anaerobic product formation in resting cells of Lactococcus lactis. Appl Environ Microbiol 70:5477–5484

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Radoš D, Turner DL, Fonseca LL, Carvalho AL, Blombach B, Eikmanns BJ, Neves AR, Santos H (2014) Carbon flux analysis by 13C nuclear magnetic resonance to determine the effect of CO2 on anaerobic succinate production by Corynebacterium glutamicum. Appl Environ Microbiol 80:3015–3024

    Article  PubMed Central  PubMed  Google Scholar 

  • Sambrook J, Fritsh E, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Tsuchida Y, Kimura S, Suzuki N, Inui M, Yukawa H (2009) Characterization of a 24-kb plasmid pCGR2 newly isolated from Corynebacterium glutamicum. Appl Microbiol Biotechnol 87:1855–1866

    Article  Google Scholar 

  • Tsuge Y, Yamamoto S, Suda M, Inui M, Yukawa H (2013) Reactions upstream of glycerate-1,3-bisphosphate drive Corynebacterium glutamicum d-lactate productivity under oxygen deprivation. Appl Microbiol Biotechnol 97:6693–6703

    Article  CAS  PubMed  Google Scholar 

  • Yamamoto S, Gunji W, Suzuki H, Toda H, Suda M, Jojima T, Inui M, Yukawa H (2012) Overexpression of genes encoding glycolytic enzymes in Corynebacterium glutamicum enhances glucose metabolism and alanine production under oxygen deprivation conditions. Appl Environ Microbiol 78:4447–4457

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yamamoto S, Suda M, Niimi S, Inui M, Yukawa H (2013) Strain optimization for efficient isobutanol production using Corynebacterium glutamicum under oxygen deprivation. Biotechnol Bioeng 110:2938–2948

    Article  CAS  PubMed  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

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a grant from the New Energy and Industrial Technology Development Organization (NEDO). This study was also partially supported by a Grant-in-Aid for Young Scientists (B) to YT from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.

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We declare that we have no conflict of interest.

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Authors and Affiliations

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

    Yota Tsuge, Kimio Uematsu, Shogo Yamamoto, Masako Suda, Hideaki Yukawa & Masayuki Inui

  2. Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5, Takayama, Ikoma, Nara, 630-0101, Japan

    Masayuki Inui

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  1. Yota Tsuge
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  2. Kimio Uematsu
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  3. Shogo Yamamoto
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  4. Masako Suda
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  5. Hideaki Yukawa
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  6. Masayuki Inui
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Correspondence to Masayuki Inui.

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Tsuge, Y., Uematsu, K., Yamamoto, S. et al. Glucose consumption rate critically depends on redox state in Corynebacterium glutamicum under oxygen deprivation. Appl Microbiol Biotechnol 99, 5573–5582 (2015). https://doi.org/10.1007/s00253-015-6540-2

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  • DOI: https://doi.org/10.1007/s00253-015-6540-2

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