Grogan, Gideon James orcid.org/0000-0003-1383-7056, Cuetos, Anibal, Danesh-Azari, Hamid-Reza et al. (4 more authors) (2020) Mutational Analysis of Linalool Dehydratase-Isomerase (LinD) Suggests Alcohol and Alkene Transformations are Catalyzed Using Non-Covalent Mechanisms. ACS Catalysis. pp. 1-11. ISSN 2155-5435
Abstract
The interconversion of non-activated alkenes and alcohols, catalyzed by (de)hydratases, has great potential in biotechnology for the generation of fine and bulk chemicals. LinD is a cofactor-independent enzyme that catalyzes the reversible (de)hydration of the tertiary alcohol (S)-linalool to the triene beta-myrcene, and also its isomerization to the primary alcohol geraniol. Structure-informed mutagenesis of LinD, followed by activity studies, confirmed essential roles for residues C171, C180 and H129 in water activation for the hydration of beta-myrcene to linalool. However, no evidence of covalent thioterpene intermediates was found using either X-ray crystallography, mass spectrometry, or QM/MM nudged elastic band simula-tions. Labelling and NMR experiments confirmed a role for residue D39 in (de)protonation of the linalool carbon C10 in the isomerization of linalool to geraniol and also the intermediacy of beta-myrcene in this isomerization reaction. X-ray, molecular dynamics and activity studies also suggested a significant role in catalysis for a mobile methionine residue M125, which exists in substantially altered orientations in different mutant structures.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2020 American Chemical Society. This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details. |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Chemistry (York) |
Funding Information: | Funder Grant number BBSRC (BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL) BB/T017805/1 |
Depositing User: | Pure (York) |
Date Deposited: | 16 Sep 2020 12:00 |
Last Modified: | 07 Feb 2025 00:29 |
Published Version: | https://doi.org/10.1021/acscatal.0c02958 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.1021/acscatal.0c02958 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:165637 |