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Engineering Soluble Methane Monooxygenase for Biocatalysis

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Methane Biocatalysis: Paving the Way to Sustainability

Abstract

Soluble methane monooxygenase (sMMO) has more than 100 known substrates in addition to its natural substrate methane. It is one of the most versatile and powerful biological oxidation catalysts, although regioselectivity and enantioselectivity with the wild-type enzyme are generally low. Protein engineering of sMMO has presented a major challenge because attempts to express the active site-containing hydroxylase component of the enzyme in Escherichia coli have to date been unsuccessful. Use of a homologous expression system, in which the enzyme is expressed in a methane-oxidising bacterium where the chromosomal copy of the sMMO genes is deleted, has allowed construction, expression and purification of active mutant enzymes. This work has given the first indications for the roles of specific amino acids in the hydroxylase component of sMMO in substrate oxidation and control of regioselectivity. Most recently, an enzyme with significantly improved activity and regioselectivity with a diaromatic substrate has been prepared. It is hoped that future work will produce recombinant sMMO derivatives developed for the production of high-value fine and bulk chemicals.

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Acknowledgements

TJS gratefully acknowledges funding for work on expression and mutagenesis of sMMO from the Biotechnology and Biological Sciences Research Council and the Biomolecular Sciences Research Centre at Sheffield Hallam University.

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Authors and Affiliations

  1. Biomolecular Sciences Research Centre, Sheffield Hallam University, Sheffield, UK

    Thomas J. Smith & Tim Nichol

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  1. Thomas J. Smith
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  2. Tim Nichol
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Correspondence to Thomas J. Smith .

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Editors and Affiliations

  1. Department of Biology, San Diego State University, San Diego, California, USA

    Marina G. Kalyuzhnaya

  2. Department of Chemical Engineering, Tsinghua University, Beijing, China

    Xin-Hui Xing

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Smith, T.J., Nichol, T. (2018). Engineering Soluble Methane Monooxygenase for Biocatalysis. In: Kalyuzhnaya, M., Xing, XH. (eds) Methane Biocatalysis: Paving the Way to Sustainability. Springer, Cham. https://doi.org/10.1007/978-3-319-74866-5_10

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