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Title: Heterologous production of an energy-conserving carbon monoxide dehydrogenase complex in the hyperthermophile Pyrococcus furiosus

Abstract

In this study, carbon monoxide (CO) is an important intermediate in anaerobic carbon fixation pathways in acetogenesis and methanogenesis. In addition, some anaerobes can utilize CO as an energy source. In the hyperthermophilic archaeon Thermococcus onnurineus, which grows optimally at 80°C, CO oxidation and energy conservation is accomplished by a respiratory complex encoded by a 16-gene cluster containing a CO dehydrogenase, a membrane-bound [NiFe]-hydrogenase and a Na +/H + antiporter module. This complex oxidizes CO, evolves CO 2 and H 2, and generates a Na+ motive force that is used to conserve energy by a Na+-dependent ATP synthase. Herein we used a bacterial artificial chromosome to insert the 13.2 kb gene cluster encoding the CO-oxidizing respiratory complex of T. onnurineus into the genome of the heterotrophic archaeon, Pyrococcus furiosus, which grows optimally at 100° C. P. furiosus is normally unable to utilize CO, however, the recombinant strain readily oxidized CO and generated H 2 at 80° C. Moreover, CO also served as an energy source and allowed the P. furiosus strain to grow with a limiting concentration of sugar or with peptides as the carbon source. Moreover, CO oxidation by P. furiosus was also coupled to the re-utilization, presumably formore » biosynthesis, of acetate generated by fermentation. The functional transfer of CO utilization between Thermococcus and Pyrococcus species demonstrated herein is representative of the horizontal gene transfer of an environmentally relevant metabolic capability. The transfer of CO utilizing, hydrogen-producing genetic modules also has applications for biohydrogen production and a CO-based industrial platform for various thermophilic organisms.« less

Authors:
 [1];  [1];  [1];  [2];  [1]
  1. Univ. of Georgia, Athens, GA (United States)
  2. North Carolina State Univ., Raleigh, NC (United States)
Publication Date:
Research Org.:
Montana State Univ., Bozeman, MT (United States); Energy Frontier Research Centers (EFRC) (United States). Center for Biological Electron Transfer and Catalysis (BETCy)
Sponsoring Org.:
USDOE
OSTI Identifier:
1242375
Grant/Contract Number:  
SC0012518
Resource Type:
Accepted Manuscript
Journal Name:
Frontiers in Microbiology
Additional Journal Information:
Journal Volume: 7; Journal ID: ISSN 1664-302X
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; hyperthermophile; archaea; hydrogen; carbon monoxide; energy; anaerobic respiration; Thermococcales

Citation Formats

Schut, Gerrit J., Lipscomb, Gina L., Nguyen, Diep M. N., Kelly, Robert M., and Adams, Michael W. W. Heterologous production of an energy-conserving carbon monoxide dehydrogenase complex in the hyperthermophile Pyrococcus furiosus. United States: N. p., 2016. Web. doi:10.3389/fmicb.2016.00029.
Schut, Gerrit J., Lipscomb, Gina L., Nguyen, Diep M. N., Kelly, Robert M., & Adams, Michael W. W. Heterologous production of an energy-conserving carbon monoxide dehydrogenase complex in the hyperthermophile Pyrococcus furiosus. United States. doi:10.3389/fmicb.2016.00029.
Schut, Gerrit J., Lipscomb, Gina L., Nguyen, Diep M. N., Kelly, Robert M., and Adams, Michael W. W. Fri . "Heterologous production of an energy-conserving carbon monoxide dehydrogenase complex in the hyperthermophile Pyrococcus furiosus". United States. doi:10.3389/fmicb.2016.00029. https://www.osti.gov/servlets/purl/1242375.
@article{osti_1242375,
title = {Heterologous production of an energy-conserving carbon monoxide dehydrogenase complex in the hyperthermophile Pyrococcus furiosus},
author = {Schut, Gerrit J. and Lipscomb, Gina L. and Nguyen, Diep M. N. and Kelly, Robert M. and Adams, Michael W. W.},
abstractNote = {In this study, carbon monoxide (CO) is an important intermediate in anaerobic carbon fixation pathways in acetogenesis and methanogenesis. In addition, some anaerobes can utilize CO as an energy source. In the hyperthermophilic archaeon Thermococcus onnurineus, which grows optimally at 80°C, CO oxidation and energy conservation is accomplished by a respiratory complex encoded by a 16-gene cluster containing a CO dehydrogenase, a membrane-bound [NiFe]-hydrogenase and a Na+/H+ antiporter module. This complex oxidizes CO, evolves CO2 and H2, and generates a Na+ motive force that is used to conserve energy by a Na+-dependent ATP synthase. Herein we used a bacterial artificial chromosome to insert the 13.2 kb gene cluster encoding the CO-oxidizing respiratory complex of T. onnurineus into the genome of the heterotrophic archaeon, Pyrococcus furiosus, which grows optimally at 100° C. P. furiosus is normally unable to utilize CO, however, the recombinant strain readily oxidized CO and generated H2 at 80° C. Moreover, CO also served as an energy source and allowed the P. furiosus strain to grow with a limiting concentration of sugar or with peptides as the carbon source. Moreover, CO oxidation by P. furiosus was also coupled to the re-utilization, presumably for biosynthesis, of acetate generated by fermentation. The functional transfer of CO utilization between Thermococcus and Pyrococcus species demonstrated herein is representative of the horizontal gene transfer of an environmentally relevant metabolic capability. The transfer of CO utilizing, hydrogen-producing genetic modules also has applications for biohydrogen production and a CO-based industrial platform for various thermophilic organisms.},
doi = {10.3389/fmicb.2016.00029},
journal = {Frontiers in Microbiology},
number = ,
volume = 7,
place = {United States},
year = {2016},
month = {1}
}

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Works referenced in this record:

Key Role for Sulfur in Peptide Metabolism and in Regulation of Three Hydrogenases in the Hyperthermophilic Archaeon Pyrococcus furiosus
journal, January 2001


H2 production from CO, formate or starch using the hyperthermophilic archaeon, Thermococcus onnurineus
journal, September 2011


Single gene insertion drives bioalcohol production by a thermophilic archaeon
journal, November 2014

  • Basen, Mirko; Schut, Gerrit J.; Nguyen, Diep M.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 49
  • DOI: 10.1073/pnas.1413789111

Bacterial synthesis gas (syngas) fermentation
journal, July 2013


A comparison of volcanic with other fluxes of atmospheric trace gas constituents
journal, January 1980


Geochemistry of high H2 and CH4 vent fluids issuing from ultramafic rocks at the Rainbow hydrothermal field (36°14′N, MAR)
journal, November 2002


Metabolic versatility in methanogens
journal, October 2014


Crystal Structure of a Carbon Monoxide Dehydrogenase Reveals a [Ni-4Fe-5S] Cluster
journal, August 2001


Physiology of the thermophilic acetogen Moorella thermoacetica
journal, December 2004


Pyrococcus furiosus sp. nov. represents a novel genus of marine heterotrophic archaebacteria growing optimally at 100°C
journal, June 1986

  • Fiala, Gerhard; Stetter, Karl O.
  • Archives of Microbiology, Vol. 145, Issue 1, p. 56-61
  • DOI: 10.1007/BF00413027

Biological conversion of carbon dioxide and hydrogen into liquid fuels and industrial chemicals
journal, June 2013

  • Hawkins, Aaron S.; McTernan, Patrick M.; Lian, Hong
  • Current Opinion in Biotechnology, Vol. 24, Issue 3, p. 376-384
  • DOI: 10.1016/j.copbio.2013.02.017

Deep Conversion of Carbon Monoxide to Hydrogen and Formation of Acetate by the Anaerobic Thermophile Carboxydothermus hydrogenoformans
journal, January 2011

  • Henstra, Anne M.; Stams, Alfons J. M.
  • International Journal of Microbiology, Vol. 2011
  • DOI: 10.1155/2011/641582

Exploiting microbial hyperthermophilicity to produce an industrial chemical, using hydrogen and carbon dioxide
journal, March 2013

  • Keller, M. W.; Schut, G. J.; Lipscomb, G. L.
  • Proceedings of the National Academy of Sciences, Vol. 110, Issue 15, p. 5840-5845
  • DOI: 10.1073/pnas.1222607110

CO-Dependent H 2 Production by Genetically Engineered Thermococcus onnurineus NA1
journal, January 2013

  • Kim, Min-Sik; Bae, Seung Seob; Kim, Yun Jae
  • Applied and Environmental Microbiology, Vol. 79, Issue 6
  • DOI: 10.1128/AEM.03298-12

Formate-driven growth coupled with H2 production
journal, September 2010

  • Kim, Yun Jae; Lee, Hyun Sook; Kim, Eun Sook
  • Nature, Vol. 467, Issue 7313
  • DOI: 10.1038/nature09375

Crystallographic Snapshots of Cyanide- and Water-Bound C-Clusters from Bifunctional Carbon Monoxide Dehydrogenase/Acetyl-CoA Synthase ,
journal, August 2009

  • Kung, Yan; Doukov, Tzanko I.; Seravalli, Javier
  • Biochemistry, Vol. 48, Issue 31
  • DOI: 10.1021/bi900574h

Thermodynamics of Formate-Oxidizing Metabolism and Implications for H 2 Production
journal, August 2012

  • Lim, Jae Kyu; Bae, Seung Seob; Kim, Tae Wan
  • Applied and Environmental Microbiology, Vol. 78, Issue 20
  • DOI: 10.1128/AEM.01316-12

Identification of a Novel Class of Membrane-Bound [NiFe]-Hydrogenases in Thermococcus onnurineus NA1 by In Silico Analysis
journal, July 2010

  • Lim, J. K.; Kang, S. G.; Lebedinsky, A. V.
  • Applied and Environmental Microbiology, Vol. 76, Issue 18
  • DOI: 10.1128/AEM.00123-10

Energy conservation by oxidation of formate to carbon dioxide and hydrogen via a sodium ion current in a hyperthermophilic archaeon
journal, July 2014

  • Lim, J. K.; Mayer, F.; Kang, S. G.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 31
  • DOI: 10.1073/pnas.1407056111

Engineering Hydrogen Gas Production from Formate in a Hyperthermophile by Heterologous Production of an 18-Subunit Membrane-bound Complex
journal, December 2013

  • Lipscomb, Gina L.; Schut, Gerrit J.; Thorgersen, Michael P.
  • Journal of Biological Chemistry, Vol. 289, Issue 5
  • DOI: 10.1074/jbc.M113.530725

Natural Competence in the Hyperthermophilic Archaeon Pyrococcus furiosus Facilitates Genetic Manipulation: Construction of Markerless Deletions of Genes Encoding the Two Cytoplasmic Hydrogenases
journal, February 2011

  • Lipscomb, Gina L.; Stirrett, Karen; Schut, Gerrit J.
  • Applied and Environmental Microbiology, Vol. 77, Issue 7, p. 2232-2238
  • DOI: 10.1128/AEM.02624-10

Purification and Properties of Acetyl-CoA Synthetase (ADP-forming), an Archaeal Enzyme of Acetate Formation and ATP Synthesis, from the Hyperthermophile Pyrococcus furiosus
journal, March 1997


Intact Functional Fourteen-subunit Respiratory Membrane-bound [NiFe]-Hydrogenase Complex of the Hyperthermophilic Archaeon Pyrococcus furiosus
journal, May 2014

  • McTernan, Patrick M.; Chandrayan, Sanjeev K.; Wu, Chang-Hao
  • Journal of Biological Chemistry, Vol. 289, Issue 28
  • DOI: 10.1074/jbc.M114.567255

Purification and catalytic properties of Ech hydrogenase from Methanosarcina barkeri
journal, October 1999


Carbon monoxide-dependent energy metabolism in anaerobic bacteria and archaea
journal, June 2008


One-carbon substrate-based biohydrogen production: Microbes, mechanism, and productivity
journal, January 2015


A simple energy-conserving system: Proton reduction coupled to proton translocation
journal, June 2003

  • Sapra, R.; Bagramyan, K.; Adams, M. W. W.
  • Proceedings of the National Academy of Sciences, Vol. 100, Issue 13, p. 7545-7550
  • DOI: 10.1073/pnas.1331436100

Autotrophy at the thermodynamic limit of life: a model for energy conservation in acetogenic bacteria
journal, November 2014

  • Schuchmann, Kai; Müller, Volker
  • Nature Reviews Microbiology, Vol. 12, Issue 12
  • DOI: 10.1038/nrmicro3365

Characterization of Ten Heterotetrameric NDP-Dependent Acyl-CoA Synthetases of the Hyperthermophilic Archaeon Pyrococcus furiosus
journal, January 2014

  • Scott, Joseph W.; Poole, Farris L.; Adams, Michael W. W.
  • Archaea, Vol. 2014
  • DOI: 10.1155/2014/176863

CO-dependent H2 evolution by Rhodospirillum rubrum: Role of CODH:CooF complex
journal, December 2006

  • Singer, Steven W.; Hirst, Marissa B.; Ludden, Paul W.
  • Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1757, Issue 12
  • DOI: 10.1016/j.bbabio.2006.10.003

Purification and catalytic properties of a CO-oxidizing:H2-evolving enzyme complex from Carboxydothermus hydrogenoformans: CO-oxidizing:H2-evolving enzyme complex
journal, October 2002


Diversity and ecophysiological features of thermophilic carboxydotrophic anaerobes: Thermophilic carboxydotrophic anaerobes
journal, May 2009


The first evidence of anaerobic CO oxidation coupled with H2 production by a hyperthermophilic archaeon isolated from a deep-sea hydrothermal vent
journal, May 2004

  • Sokolova, Tatyana G.; Jeanthon, Christian; Kostrikina, Nadezhda A.
  • Extremophiles, Vol. 8, Issue 4
  • DOI: 10.1007/s00792-004-0389-0

Utilization of energy for growth and maintenance in continuous and batch cultures of microorganisms
journal, February 1973

  • Stouthamer, A. H.; Bettenhaussen, Corry
  • Biochimica et Biophysica Acta (BBA) - Reviews on Bioenergetics, Vol. 301, Issue 1
  • DOI: 10.1016/0304-4173(73)90012-8

Carboxydothermus hydrogenoformans gen. nov., sp. nov., a CO-utilizing Thermophilic Anaerobic Bacterium from Hydrothermal Environments of Kunashir Island
journal, July 1991


A functional Ni-Ni-[4Fe-4S] cluster in the monomeric acetyl-CoA synthase from Carboxydothermus hydrogenoformans
journal, December 2003

  • Svetlitchnyi, V.; Dobbek, H.; Meyer-Klaucke, W.
  • Proceedings of the National Academy of Sciences, Vol. 101, Issue 2
  • DOI: 10.1073/pnas.0304262101

Evidence for Horizontal Gene Transfer of Anaerobic Carbon Monoxide Dehydrogenases
journal, January 2012

  • Techtmann, Stephen M.; Lebedinsky, Alexander V.; Colman, Albert S.
  • Frontiers in Microbiology, Vol. 3
  • DOI: 10.3389/fmicb.2012.00132

Energy metabolism of methanogenic bacteria
journal, July 1990


Anaerobic growth of a Rhodopseudomonas species in the dark with carbon monoxide as sole carbon and energy substrate.
journal, September 1976


Life with CO or CO2 and H2 as a source of carbon and energy.
journal, February 1991


Life in Hot Carbon Monoxide: The Complete Genome Sequence of Carboxydothermus hydrogenoformans Z-2901
journal, January 2005


Parallel evolution of transcriptome architecture during genome reorganization
journal, July 2011

  • Yoon, S. H.; Reiss, D. J.; Bare, J. C.
  • Genome Research, Vol. 21, Issue 11, p. 1892-1904
  • DOI: 10.1101/gr.122218.111

Proteome analysis of Thermococcus onnurineus NA1 reveals the expression of hydrogen gene cluster under carboxydotrophic growth
journal, September 2011