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A catalytically active [Mn]-hydrogenase incorporating a non-native metal cofactor


Nature carefully selects specific metal ions for incorporation into the enzymes that catalyse the chemical reactions necessary for life. Hydrogenases, enzymes that activate molecular H2, exclusively utilize Ni and Fe in [NiFe]-, [FeFe]- and [Fe]-hydrogeanses. However, other transition metals are known to activate or catalyse the production of hydrogen in synthetic systems. Here, we report the development of a biomimetic model complex of [Fe]-hydrogenase that incorporates a Mn, as opposed to a Fe, metal centre. This Mn complex is able to heterolytically cleave H2 as well as catalyse hydrogenation reactions. The incorporation of the model into an apoenzyme of [Fe]-hydrogenase results in a [Mn]-hydrogenase with an enhanced occupancy-normalized activity over an analogous semi-synthetic [Fe]-hydrogenase. These findings demonstrate a non-native metal hydrogenase that shows catalytic functionality and that hydrogenases based on a manganese active site are viable.

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Fig. 1: Active site of [Fe]-hydrogenase and synthesis, structure and catalytic activity of its Mn models.
Fig. 2: Computational study of the mechanism of hydrogenation.
Fig. 3: Activity of the semi-synthetic [Mn]-hydrogenase.

Data availability

The authors declare that the data supporting the findings of this study are available from the corresponding authors upon reasonable request. CCDC-1856616 and CCDC-1856615 contain the supplementary crystallographic data for 3 and 4(18-crown-6), respectively. These data can be obtained free of charge from The Cambridge Crystallographic Data Centre via


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This work was supported by the Swiss National Science Foundation (to X.L.H.), European Union Marie Sklodowska-Curie Individual Fellowships (794000 to H.-J.P.), Max Planck Society (to S.S.) and Deutsche Forschungsgemeinschaft (SH 87/1-1, to S.S.). M.D.W. acknowledges C. Corminboeuf (EPFL)for financial support and the Laboratory for Computational Molecular Design (EPFL) for providing computing resources. G.H. was supported by a fellowship from the China Scholarship Council (CSC).

Author information




S.S. and X.L.H. directed the research. H.-J.P. conducted the experiments of the Mn complexes; G.F.H. conducted experiments of the semi-synthetic enzymes; M.D.W. did the computations; F.F.T. did the crystallographic study of Mn complexes; K.A. performed infrared spectroscopy of the enzymes; All authors discussed the data. X.L.H. wrote the manuscript with contributions from H.-J.P and S.S.

Corresponding authors

Correspondence to Seigo Shima or Xile Hu.

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The authors declare no competing interests.

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Supplementary Information

Supplementary Information

Supplementary experimental procedures, experimental data, optimization data, compounds characterization data, and computational methods

Cif file for complex 3

Crystallographic file for complex 3; CCDC 1856616

Cif file for complex 4

Crystallographic file for complex 4(18-crown-6); CCDC 1856615

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Pan, HJ., Huang, G., Wodrich, M.D. et al. A catalytically active [Mn]-hydrogenase incorporating a non-native metal cofactor. Nat. Chem. 11, 669–675 (2019).

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