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Title: Light-driven carbon dioxide reduction to methane by nitrogenase in a photosynthetic bacterium

Nitrogenase is an ATP-requiring enzyme capable of carrying out multielectron reductions of inert molecules. A purified remodeled nitrogenase containing two amino acid substitutions near the site of its FeMo cofactor was recently described as having the capacity to reduce carbon dioxide (CO 2) to methane (CH 4). Here, we developed the anoxygenic phototroph, Rhodopseudomonas palustris, as a biocatalyst capable of light-driven CO 2 reduction to CH 4 in vivo using this remodeled nitrogenase. Conversion of CO 2 to CH 4 by R. palustris required constitutive expression of nitrogenase, which was achieved by using a variant of the transcription factor NifA that is able to activate expression of nitrogenase under all growth conditions. Also, light was required for generation of ATP by cyclic photophosphorylation. CH 4 production by R. palustris could be controlled by manipulating the distribution of electrons and energy available to nitrogenase. Furthermore, this work shows the feasibility of using microbes to generate hydrocarbons from CO 2 in one enzymatic step using light energy.
Authors:
 [1] ;  [1] ;  [2] ;  [2] ;  [2] ;  [3] ;  [2] ; ORCiD logo [1]
  1. Univ. of Washington, Seattle, WA (United States)
  2. Utah State Univ., Logan, UT (United States)
  3. Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States)
Publication Date:
Grant/Contract Number:
SC0012518
Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 113; Journal Issue: 36; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Research Org:
Utah State Univ., Logan, UT (United States)
Sponsoring Org:
USDOE
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; nitrogenase; Rhodopseudomonas; bioenergy; methane; engineered bacterium
OSTI Identifier:
1319953
Alternate Identifier(s):
OSTI ID: 1436487

Fixen, Kathryn R., Zheng, Yanning, Harris, Derek F., Shaw, Sudipta, Yang, Zhi -Yong, Dean, Dennis R., Seefeldt, Lance C., and Harwood, Caroline S.. Light-driven carbon dioxide reduction to methane by nitrogenase in a photosynthetic bacterium. United States: N. p., Web. doi:10.1073/pnas.1611043113.
Fixen, Kathryn R., Zheng, Yanning, Harris, Derek F., Shaw, Sudipta, Yang, Zhi -Yong, Dean, Dennis R., Seefeldt, Lance C., & Harwood, Caroline S.. Light-driven carbon dioxide reduction to methane by nitrogenase in a photosynthetic bacterium. United States. doi:10.1073/pnas.1611043113.
Fixen, Kathryn R., Zheng, Yanning, Harris, Derek F., Shaw, Sudipta, Yang, Zhi -Yong, Dean, Dennis R., Seefeldt, Lance C., and Harwood, Caroline S.. 2016. "Light-driven carbon dioxide reduction to methane by nitrogenase in a photosynthetic bacterium". United States. doi:10.1073/pnas.1611043113.
@article{osti_1319953,
title = {Light-driven carbon dioxide reduction to methane by nitrogenase in a photosynthetic bacterium},
author = {Fixen, Kathryn R. and Zheng, Yanning and Harris, Derek F. and Shaw, Sudipta and Yang, Zhi -Yong and Dean, Dennis R. and Seefeldt, Lance C. and Harwood, Caroline S.},
abstractNote = {Nitrogenase is an ATP-requiring enzyme capable of carrying out multielectron reductions of inert molecules. A purified remodeled nitrogenase containing two amino acid substitutions near the site of its FeMo cofactor was recently described as having the capacity to reduce carbon dioxide (CO2) to methane (CH4). Here, we developed the anoxygenic phototroph, Rhodopseudomonas palustris, as a biocatalyst capable of light-driven CO2 reduction to CH4 in vivo using this remodeled nitrogenase. Conversion of CO2 to CH4 by R. palustris required constitutive expression of nitrogenase, which was achieved by using a variant of the transcription factor NifA that is able to activate expression of nitrogenase under all growth conditions. Also, light was required for generation of ATP by cyclic photophosphorylation. CH4 production by R. palustris could be controlled by manipulating the distribution of electrons and energy available to nitrogenase. Furthermore, this work shows the feasibility of using microbes to generate hydrocarbons from CO2 in one enzymatic step using light energy.},
doi = {10.1073/pnas.1611043113},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 36,
volume = 113,
place = {United States},
year = {2016},
month = {8}
}

Works referenced in this record:

A Broad Host Range Mobilization System for In Vivo Genetic Engineering Transposon Mutagenesis in Gram Negative Bacteria
journal, November 1983
  • Simon, R.; Priefer, U.; Pühler, A.
  • Bio/Technology, Vol. 1, Issue 9, p. 784-791
  • DOI: 10.1038/nbt1183-784