The [NiFe]-Hydrogenase of Pyrococcus furiosus Exhibits a New Type of Oxygen Tolerance

Patrick Kwan, Chelsea L. McIntosh, David P. Jennings, R. Chris Hopkins, Sanjeev K. Chandrayan, Chang Hao Wu, Michael W W Adams, Anne Jones

Research output: Contribution to journalArticle

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Abstract

We report the first direct electrochemical characterization of the impact of oxygen on the hydrogen oxidation activity of an oxygen-tolerant, group 3, soluble [NiFe]-hydrogenase: hydrogenase I from Pyrococcus furiosus (PfSHI), which grows optimally near 100 °C. Chronoamperometric experiments were used to probe the sensitivity of PfSHI hydrogen oxidation activity to both brief and prolonged exposure to oxygen. For experiments between 15 and 80 °C, following short (2 under oxidizing conditions, PfSHI always maintains some fraction of its initial hydrogen oxidation activity; i.e., it is oxygen-tolerant. Reactivation experiments show that two inactive states are formed by interaction with oxygen and both can be quickly (2/ 99% H2 exposure, the H2oxidation activity drops nearly to zero. However, at 80 °C, up to 32% of the enzymes oxidation activity is retained. Reactivation of PfSHI following sustained exposure to oxygen occurs on a much longer time scale (tens of minutes), suggesting that a third inactive species predominates under these conditions. These results stand in contrast to the properties of oxygen-tolerant, group 1 [NiFe]-hydrogenases, which form a single state upon reaction with oxygen, and we propose that this new type of hydrogenase should be referred to as oxygen-resilient. Furthermore, PfSHI, like other group 3 [NiFe]-hydrogenases, does not possess the proximal [4Fe3S] cluster associated with the oxygen tolerance of some group 1 enzymes. Thus, a new mechanism is necessary to explain the observed oxygen tolerance in soluble, group 3 [NiFe]-hydrogenases, and we present a model integrating both electrochemical and spectroscopic results to define the relationships of these inactive states.

Original languageEnglish (US)
Pages (from-to)13556-13565
Number of pages10
JournalJournal of the American Chemical Society
Volume137
Issue number42
DOIs
StatePublished - Oct 28 2015

Fingerprint

Pyrococcus furiosus
Oxygen
Hydrogenase
Hydrogen
Oxidation
nickel-iron hydrogenase
Enzymes
Experiments

ASJC Scopus subject areas

  • Chemistry(all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Kwan, P., McIntosh, C. L., Jennings, D. P., Hopkins, R. C., Chandrayan, S. K., Wu, C. H., ... Jones, A. (2015). The [NiFe]-Hydrogenase of Pyrococcus furiosus Exhibits a New Type of Oxygen Tolerance. Journal of the American Chemical Society, 137(42), 13556-13565. https://doi.org/10.1021/jacs.5b07680

The [NiFe]-Hydrogenase of Pyrococcus furiosus Exhibits a New Type of Oxygen Tolerance. / Kwan, Patrick; McIntosh, Chelsea L.; Jennings, David P.; Hopkins, R. Chris; Chandrayan, Sanjeev K.; Wu, Chang Hao; Adams, Michael W W; Jones, Anne.

In: Journal of the American Chemical Society, Vol. 137, No. 42, 28.10.2015, p. 13556-13565.

Research output: Contribution to journalArticle

Kwan, P, McIntosh, CL, Jennings, DP, Hopkins, RC, Chandrayan, SK, Wu, CH, Adams, MWW & Jones, A 2015, 'The [NiFe]-Hydrogenase of Pyrococcus furiosus Exhibits a New Type of Oxygen Tolerance', Journal of the American Chemical Society, vol. 137, no. 42, pp. 13556-13565. https://doi.org/10.1021/jacs.5b07680
Kwan P, McIntosh CL, Jennings DP, Hopkins RC, Chandrayan SK, Wu CH et al. The [NiFe]-Hydrogenase of Pyrococcus furiosus Exhibits a New Type of Oxygen Tolerance. Journal of the American Chemical Society. 2015 Oct 28;137(42):13556-13565. https://doi.org/10.1021/jacs.5b07680
Kwan, Patrick ; McIntosh, Chelsea L. ; Jennings, David P. ; Hopkins, R. Chris ; Chandrayan, Sanjeev K. ; Wu, Chang Hao ; Adams, Michael W W ; Jones, Anne. / The [NiFe]-Hydrogenase of Pyrococcus furiosus Exhibits a New Type of Oxygen Tolerance. In: Journal of the American Chemical Society. 2015 ; Vol. 137, No. 42. pp. 13556-13565.
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AB - We report the first direct electrochemical characterization of the impact of oxygen on the hydrogen oxidation activity of an oxygen-tolerant, group 3, soluble [NiFe]-hydrogenase: hydrogenase I from Pyrococcus furiosus (PfSHI), which grows optimally near 100 °C. Chronoamperometric experiments were used to probe the sensitivity of PfSHI hydrogen oxidation activity to both brief and prolonged exposure to oxygen. For experiments between 15 and 80 °C, following short (2 under oxidizing conditions, PfSHI always maintains some fraction of its initial hydrogen oxidation activity; i.e., it is oxygen-tolerant. Reactivation experiments show that two inactive states are formed by interaction with oxygen and both can be quickly (2/ 99% H2 exposure, the H2oxidation activity drops nearly to zero. However, at 80 °C, up to 32% of the enzymes oxidation activity is retained. Reactivation of PfSHI following sustained exposure to oxygen occurs on a much longer time scale (tens of minutes), suggesting that a third inactive species predominates under these conditions. These results stand in contrast to the properties of oxygen-tolerant, group 1 [NiFe]-hydrogenases, which form a single state upon reaction with oxygen, and we propose that this new type of hydrogenase should be referred to as oxygen-resilient. Furthermore, PfSHI, like other group 3 [NiFe]-hydrogenases, does not possess the proximal [4Fe3S] cluster associated with the oxygen tolerance of some group 1 enzymes. Thus, a new mechanism is necessary to explain the observed oxygen tolerance in soluble, group 3 [NiFe]-hydrogenases, and we present a model integrating both electrochemical and spectroscopic results to define the relationships of these inactive states.

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