Enhanced Hydrogen Production by Co-cultures of Hydrogenase and Nitrogenase in Escherichia coli


Rhodobacter sphaeroides is a bacterium that can produce hydrogen by interaction with hydrogenase and nitrogenase. We report a hydrogen production system using co-cultivation of hydrogenase in liquid medium and immobilized nitrogenase in Escherichia coli. The recombinant plasmid has been constructed to analyze the effect of hydrogen production on the expression of hupSL hydrogenase and nifHDK nitrogenase isolated from R. sphaeroides. All recombinant E. coli strains were cultured anaerobically, and cells for nitrogenase were immobilized in agar gel, whereas cells for hydrogenase were supplemented on the nitrogenase agar gel. The hupSL hydrogenase has been observed to enhance hydrogen production and hydrogenase activity under co-culture with nifHDK nitrogenase. The maximum hydrogen production has been obtained at an agar gel concentration and a cell concentration for co-culture of 2 % and 6.4 × 108 CFU. Thus, co-culture of hupSL hydrogenase and nifHDK nitrogenase provides a promising route for enhancing the hydrogen production and hydrogenase activity.

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This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project title: Development of Target-specific Antimicrobial and Neutralizing Agents for Livestock Biological Hazardous Factors, Project No: PJ01052701)” Rural Development Administration, Republic of Korea.

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Correspondence to Yang-Hoon Kim or Jiho Min.

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Hyun Jeong Lee and Simranjeet Singh Sekhon have contributed equally to this work.

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Lee, H.J., Sekhon, S.S., Kim, Y.S. et al. Enhanced Hydrogen Production by Co-cultures of Hydrogenase and Nitrogenase in Escherichia coli . Curr Microbiol 72, 242–247 (2016). https://doi.org/10.1007/s00284-015-0941-4

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  • Hydrogen Production
  • Hydrogenase Activity
  • Dark Fermentation
  • MoFe Protein
  • Fermentative Hydrogen Production