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Oxidation of methane by a biological dicopper centre

Nature volume 465, pages 115–119 (2010)Cite this article

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Abstract

Vast world reserves of methane gas are underutilized as a feedstock for the production of liquid fuels and chemicals owing to the lack of economical and sustainable strategies for the selective oxidation of methane to methanol1. Current processes to activate the strong C–H bond (104 kcal mol-1) in methane require high temperatures, are costly and inefficient, and produce waste2. In nature, methanotrophic bacteria perform this reaction under ambient conditions using metalloenzymes called methane monooxygenases (MMOs). MMOs thus provide the optimal model for an efficient, environmentally sound catalyst3. There are two types of MMO. Soluble MMO (sMMO) is expressed by several strains of methanotroph under copper-limited conditions and oxidizes methane with a well-characterized catalytic di-iron centre4. Particulate MMO (pMMO) is an integral membrane metalloenzyme produced by all methanotrophs and is composed of three subunits, pmoA, pmoB and pmoC, arranged in a trimeric α3β3γ3 complex5. Despite 20 years of research and the availability of two crystal structures, the metal composition and location of the pMMO metal active site are not known. Here we show that pMMO activity is dependent on copper, not iron, and that the copper active site is located in the soluble domains of the pmoB subunit rather than within the membrane. Recombinant soluble fragments of pmoB (spmoB) bind copper and have propylene and methane oxidation activities. Disruption of each copper centre in spmoB by mutagenesis indicates that the active site is a dicopper centre. These findings help resolve the pMMO controversy and provide a promising new approach to developing environmentally friendly C–H oxidation catalysts.

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Figure 1: Structure of the M. capsulatus (Bath) pMMO protomer.
Figure 2: Metal analysis.
Figure 3: Restoration of activity to apo-pMMO by the addition of copper.
Figure 4: Copper EXAFS data and simulations for pMMO and spmoB variants.
Figure 5: Catalytic activity of spmoB proteins.

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Acknowledgements

This work was supported by US National Institutes of Health (NIH) grants GM070473 (A.C.R.) and DK068139 (T.L.S.). Portions of this research were carried out at the Stanford Synchrotron Radiation Lightsource (SSRL). The SSRL is a national user facility operated by Stanford University on behalf of the US Department of Energy, Office of Basic Energy Sciences. The SSRL Structural Molecular Biology Program is supported by the Department of Energy, Office of Biological and Environmental Research, and by the NIH, National Center for Research Resources, Biomedical Technology Program.

Author Contributions R.B., S.M.S., S.R. and L.A.Y. performed experiments. R.B., S.M.S., S.R., T.L.S. and A.C.R. contributed to experimental design, data analysis and manuscript preparation.

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Author notes

  1. Ramakrishnan Balasubramanian and Stephen M. Smith: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry, Molecular Biology and Cell Biology,

    Ramakrishnan Balasubramanian, Stephen M. Smith, Liliya A. Yatsunyk & Amy C. Rosenzweig

  2. Department of Chemistry, Northwestern University, Evanston, Illinois 60208, USA,

    Ramakrishnan Balasubramanian, Stephen M. Smith, Liliya A. Yatsunyk & Amy C. Rosenzweig

  3. Department of Biochemistry and Molecular Biology, Wayne State University, School of Medicine, Detroit, Michigan 48201, USA,

    Swati Rawat & Timothy L. Stemmler

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  1. Ramakrishnan Balasubramanian
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Correspondence to Timothy L. Stemmler or Amy C. Rosenzweig.

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Balasubramanian, R., Smith, S., Rawat, S. et al. Oxidation of methane by a biological dicopper centre. Nature 465, 115–119 (2010). https://doi.org/10.1038/nature08992

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