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Microbial production of ethanol from acetate by engineered Ralstonia eutropha

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Abstract

This study was performed to produce ethanol from acetate using a genetically engineered Ralstonia eutropha. In order to genetically modify R. eutropha H16, phaCAB operon encoding metabolic pathway genes from acetyl-CoA to polyhydroxybutyrate (PHB) was deleted and adhE encoding an alcohol dehydrogenase from Escherichia coli was overexpressed for conversion of acetyl-CoA to ethanol. The resulting strain produced ethanol up to 170 mg/L when cultivated in minimal media supplemented with 5 g/L of acetate as a sole carbon source. Growth and ethanol production were optimized by adjusting nitrogen source (NH4Cl) content and repetitive feeding of acetate into the bacterial culture, by which the ethanol production was reached to approximately 350 mg/L for 84 h.

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References

  1. Lee, S. K., H. Chou, T. S. Ham, T. S. Lee, and J. D. Keasling (2008) Metabolic engineering of microorganisms for biofuels production: From bugs to synthetic biology to fuels. Curr. Opin. Biotechnol. 19: 556–563.

    Article  CAS  Google Scholar 

  2. Balat, M. and H. Balat (2009) Recent trends in global production and utilization of bio-ethanol fuel. Appl. Energy 86: 2273–2282.

    Article  CAS  Google Scholar 

  3. Hira, A. (2011) Sugar rush: Prospects for a global ethanol market. Energy Policy 39: 6925–6935.

    Article  CAS  Google Scholar 

  4. Choi, Y. I., B. G. Jung, N. C. Sung, and Y. R. Han (2015) A study on the drying characteristics from mixture of food waste and sawdust by using microwave/inner-cycle thermal-air drying process. J. Mater Cycles Waste Manag. 17: 359–368.

    Article  CAS  Google Scholar 

  5. Kim, W., B. G. Ryu, S. Kim, S. W. Heo, D. Kim, J. Kim, H. Jo, J. H. Kwon, and J. W. Yang (2014) Quantitative analysis of microbial community structure in two-phase anaerobic digesters treating food wastewater. Kor. J. Chem. Eng. 31: 381–385.

    Article  CAS  Google Scholar 

  6. Pohlmann, A., W. F. Fricke, F. Reinecke, B. Kusian, H. Liesegang, R. Cramm, T. Eitinger, C. Ewering, M. Pötter, E. Schwartz, A. Strittmatter, I. Voss, G. Gottschalk, A. Steinbüchell, B. Friedrich, and B. Bowien (2006) Genome sequence of the bioplastic-producing “Knallgas” bacterium Ralstonia eutropha H16. Nat. Biotechnol. 24: 1257–1262.

    Article  Google Scholar 

  7. Jeon, J. M., C. J. Brigham, Y. H. Kim, H. J. Kim, D. H. Yi, H. Kim, C. Rha, A. J. Sinskey, and Y. H. Yang (2014) Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-co-HHx)) from butyrate using engineered Ralstonia eutropha. Appl. Microbiol. Biotechnol. 98: 5461–5469.

    Article  CAS  Google Scholar 

  8. Yang, Y. H., C. J. Brigham, C. F. Budde, P. Boccazzi, L. B. Willis, M. A. Hassan, Z. A. Yusof, C. Rha, and A. J. Sinskey (2010) Optimization of growth media components for polyhydroxyalkanoate (PHA) production from organic acids by Ralstonia eutropha. Appl. Microbiol. Biotechnol. 87: 2037–2045.

    Article  CAS  Google Scholar 

  9. Grousseau, E., J. Lu, N. Gorret, S. E. Guillouet, and A. J. Sinskey (2014) Isopropanol production with engineered Cupriavidus necator as bioproduction platform. Appl. Microbiol. Biotechnol. 98: 4277–4290.

    Article  CAS  Google Scholar 

  10. Lu, J., C. J. Brigham, C. S. Gai, and A. J. Sinskey (2012) Studies on the production of branched-chain alcohols in engineered Ralstonia eutropha. Appl. Microbiol. Biotechnol. 96: 283–297.

    Article  CAS  Google Scholar 

  11. Muller, J., D. MacEachran, H. Burd, N. Sathitsuksanoh, C. Bi, Y. C. Yeh, T. S. Lee, N. J. Hillson, S. R. Chhabra, S.W, Singer, and H. R. Beller (2013) Engineering of Ralstonia eutropha H16 for autotrophic and heterotrophic production of methyl ketones. Appl. Microbiol. Biotechnol. 79: 4433–4439.

    CAS  Google Scholar 

  12. York, G. M., J. Stubbe, and A. J. Sinskey (2001) New insight into the role of the PhaP phasin of Ralstonia eutropha in promoting synthesis of polyhydroxybutyrate. J. Bacteriol. 183: 2394–2397.

    Article  CAS  Google Scholar 

  13. Friedrich, B., C. Hogrefe, and H. G. Schlegel (1981) Naturallyoccurring genetic transfer of hydrogen-oxidizing ability between strains of Alcaligenes eutrophus. J. Bacteriol. 147: 198–205.

    CAS  Google Scholar 

  14. Gibson, D. G., L. Young, R. Y. Chuang, J. C. Venter, C. A. Hutchison, and H. O. Smith (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat. Meth. 6: 343–U341.

    Article  CAS  Google Scholar 

  15. Hong, W. K., C. H. Kim, S. Y. Heo, L. Luo, B. R. Oh, and J. W. Seo (2010) Enhanced production of ethanol from glycerol by engineered Hansenula polymorpha expressing pyruvate decarboxylase and aldehyde dehydrogenase genes from Zymomonas mobilis. Biotechnol. Lett. 32: 1077–1082.

    Article  CAS  Google Scholar 

  16. Wang, Z., M. Chen, Y. Xu, S. Li, W. Lu, S. Ping, W. Zhang, and M. Lin (2008) An ethanol-tolerant recombinant Escherichia coli expressing Zymomonas mobilis pdc and adhB genes for enhanced ethanol production from xylose. Biotechnol. Lett. 30: 657–663.

    Article  CAS  Google Scholar 

  17. Gunasekaran, P. and K. C. Raj (1999) Ethanol fermentation technology - Zymomonas mobilis. Curr. Sci. 77: 56–68.

    CAS  Google Scholar 

  18. Wang, J. and J. Yu (2001) Kinetic analysis on formation of poly(3-hydroxybutyrate) from acetic acid by Ralstonia eutropha under chemically defined conditions. J. Ind. Microbiol. Biotechnol. 26: 121–126.

    Article  CAS  Google Scholar 

  19. Axe, D. D. and J. E. Bailey (1995) Transport of lactate and acetate through the energized cytoplasmic membrane of Escherichia coli. Biotechnol. Bioeng. 47: 8–19.

    Article  CAS  Google Scholar 

  20. Trcek, J., N. P. Mira, and L. R. Jarboe (2015) Adaptation and tolerance of bacteria against acetic acid. Appl. Microbiol. Biotechnol. 99: 6215–6229.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Chemical and Biological Engineering, Korea University, Seoul, 136-713, Korea

    Hye-Mi Lee, Bo-Young Jeon & Min-Kyu Oh

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  1. Hye-Mi Lee
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  2. Bo-Young Jeon
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  3. Min-Kyu Oh
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Correspondence to Min-Kyu Oh.

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Lee, HM., Jeon, BY. & Oh, MK. Microbial production of ethanol from acetate by engineered Ralstonia eutropha . Biotechnol Bioproc E 21, 402–407 (2016). https://doi.org/10.1007/s12257-016-0197-2

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  • DOI: https://doi.org/10.1007/s12257-016-0197-2

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