Engineering Ralstonia eutropha for Production of Isobutanol from CO2, H2, and O2

  • Christopher J. Brigham
  • Claudia S. Gai
  • Jingnan Lu
  • Daan R. Speth
  • R. Mark Worden
  • Anthony J. Sinskey


Isobutanol (IBT) can be used as a 100% replacement for gasoline in existing automobile engines, has >90% of the energy density of gasoline and is compatible with established fuel distribution infrastructure. The facultatively autotrophic bacterium Ralstonia eutropha can utilize H2 for energy and CO2 for carbon and is also employed in industrial processes that produce biodegradable plastics. Using a carefully designed production pathway, R. eutropha, a genetically tractable organism, can be modified to produce biofuels from autotrophic growth. Microbial production of IBT can be achieved by directing the flow of carbon through a ­synthetic production pathway involving the branched-chain amino acid biosynthesis pathway, a heterologously expressed ketoisovalerate decarboxylase, and a broad substrate specificity alcohol dehydrogenase. We discuss the motivations and the methods used to engineer R. eutropha to produce the liquid transportation fuel IBT from CO2, H2, and O2.


Carbonic Anhydrase Carbonic Anhydrase Gene Intracellular NADPH Soluble Hydrogenase Eutropha Strain 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank John W. Quimby for critical review of this manuscript. D.S. is ­supported by the following foundations: Nijmeegs Universiteitsfonds (SNUF), Fundatie van de Vrijvrouwe van Renswoude te’s-Gravenhage, and Dr. Hendrik Muller’s Vaderlandsch Fonds. Other authors are supported, fully or in part, by the Advanced Research Projects Agency—Energy (ARPA-E) Electrofuels project. We wish to thank the ARPA-E directors and staff for their support.


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Christopher J. Brigham
    • 1
  • Claudia S. Gai
    • 1
  • Jingnan Lu
    • 2
  • Daan R. Speth
    • 1
    • 3
  • R. Mark Worden
    • 4
  • Anthony J. Sinskey
    • 1
    • 5
    • 6
  1. 1.Department of BiologyMassachusetts Institute of TechnologyCambridgeUSA
  2. 2.Department of ChemistryMassachusetts Institute of TechnologyCambridgeUSA
  3. 3.Department of Microbiology, IWWRRadboud University NijmegenNijmegenThe Netherlands
  4. 4.Department of Chemical Engineering and Materials ScienceMichigan State UniversityEast LansingUSA
  5. 5.Engineering Systems DivisionMassachusetts Institute of TechnologyCambridgeUSA
  6. 6.Health Sciences Technology DivisionMassachusetts Institute of TechnologyCambridgeUSA

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