Effects of Active‐Site Modification and Quaternary Structure on the Regioselectivity of Catechol‐O‐Methyltransferase |
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| Authors: | Dr Brian J C Law Dr Matthew R Bennett Dr Mark L Thompson Dr Colin Levy Dr Sarah A Shepherd Prof David Leys Prof Jason Micklefield |
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| Institution: | School of Chemistry & Manchester Institute of Biotechnology, University of Manchester, Manchester, UK |
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| Abstract: | Catechol‐O‐methyltransferase (COMT), an important therapeutic target in the treatment of Parkinson's disease, is also being developed for biocatalytic processes, including vanillin production, although lack of regioselectivity has precluded its more widespread application. By using structural and mechanistic information, regiocomplementary COMT variants were engineered that deliver either meta‐ or para‐methylated catechols. X‐ray crystallography further revealed how the active‐site residues and quaternary structure govern regioselectivity. Finally, analogues of AdoMet are accepted by the regiocomplementary COMT mutants and can be used to prepare alkylated catechols, including ethyl vanillin. |
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| Keywords: | biocatalysis enzyme structure methyltransferases protein engineering regioselectivity |
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