Rational Design of Resveratrol O-methyltransferase for the Production of Pinostilbene
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Title: | Rational Design of Resveratrol O-methyltransferase for the Production of Pinostilbene |
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Authors: | Herrera, Daniela P. | Chánique, Andrea M. | Martínez Márquez, Ascensión | Bru-Martinez, Roque | Kourist, Robert | Parra, Loreto P. | Schüller, Andreas |
Research Group/s: | Proteómica y Genómica Funcional de Plantas |
Center, Department or Service: | Universidad de Alicante. Departamento de Agroquímica y Bioquímica |
Keywords: | Enzyme engineering | O-methyltransferases | Pinostilbene | Protein models | Substrate selectivity | Stilbenes |
Knowledge Area: | Bioquímica y Biología Molecular |
Issue Date: | 21-Apr-2021 |
Publisher: | MDPI |
Citation: | Herrera DP, Chánique AM, Martínez-Márquez A, Bru-Martínez R, Kourist R, Parra LP, Schüller A. Rational Design of Resveratrol O-methyltransferase for the Production of Pinostilbene. International Journal of Molecular Sciences. 2021; 22(9):4345. https://doi.org/10.3390/ijms22094345 |
Abstract: | Pinostilbene is a monomethyl ether analog of the well-known nutraceutical resveratrol. Both compounds have health-promoting properties, but the latter undergoes rapid metabolization and has low bioavailability. O-methylation improves the stability and bioavailability of resveratrol. In plants, these reactions are performed by O-methyltransferases (OMTs). Few efficient OMTs that monomethylate resveratrol to yield pinostilbene have been described so far. Here, we report the engineering of a resveratrol OMT from Vitis vinifera (VvROMT), which has the highest catalytic efficiency in di-methylating resveratrol to yield pterostilbene. In the absence of a crystal structure, we constructed a three-dimensional protein model of VvROMT and identified four critical binding site residues by applying different in silico approaches. We performed point mutations in these positions generating W20A, F24A, F311A, and F318A variants, which greatly reduced resveratrol’s enzymatic conversion. Then, we rationally designed eight variants through comparison of the binding site residues with other stilbene OMTs. We successfully modified the native substrate selectivity of VvROMT. Variant L117F/F311W showed the highest conversion to pinostilbene, and variant L117F presented an overall increase in enzymatic activity. Our results suggest that VvROMT has potential for the tailor-made production of stilbenes. |
Sponsor: | This research was funded by PROYECTO INTERDISCIPLINA-VRI-UC-II160020, number 3514-913, Pontificia Universidad Católica de Chile (to L.P.P. and A.S.) and BECA DE DOCTORADO NACIONAL 2016, number 21161084, National Agency for Research and Development (ANID), Chile (to D.P.H.), for which we are grateful. The APC was funded by Pontificia Universidad Católica de Chile. |
URI: | http://hdl.handle.net/10045/114634 |
ISSN: | 1661-6596 (Print) | 1422-0067 (Online) |
DOI: | 10.3390/ijms22094345 |
Language: | eng |
Type: | info:eu-repo/semantics/article |
Rights: | © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
Peer Review: | si |
Publisher version: | https://doi.org/10.3390/ijms22094345 |
Appears in Collections: | INV - Proteómica y Genómica Funcional de Plantas - Artículos de Revistas |
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Herrera_etal_2021_IntJMolSci.pdf | 2,59 MB | Adobe PDF | Open Preview | |
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