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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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.
Balat, M. and H. Balat (2009) Recent trends in global production and utilization of bio-ethanol fuel. Appl. Energy 86: 2273–2282.
Hira, A. (2011) Sugar rush: Prospects for a global ethanol market. Energy Policy 39: 6925–6935.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Gunasekaran, P. and K. C. Raj (1999) Ethanol fermentation technology - Zymomonas mobilis. Curr. Sci. 77: 56–68.
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.
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.
Trcek, J., N. P. Mira, and L. R. Jarboe (2015) Adaptation and tolerance of bacteria against acetic acid. Appl. Microbiol. Biotechnol. 99: 6215–6229.
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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
- metabolic engineering
- Ralstonia eutropha