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Lookup NU author(s): Professor Paul RaceORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© 2023 The Authors. ChemBioChem published by Wiley-VCH GmbH.Stereoselective carbon-carbon bond forming reactions are quintessential transformations in organic synthesis. One example is the Diels-Alder reaction, a [4+2] cycloaddition between a conjugated diene and a dienophile to form cyclohexenes. The development of biocatalysts for this reaction is paramount for unlocking sustainable routes to a plethora of important molecules. To obtain a comprehensive understanding of naturally evolved [4+2] cyclases, and to identify hitherto uncharacterised biocatalysts for this reaction, we constructed a library comprising forty-five enzymes with reported or predicted [4+2] cycloaddition activity. Thirty-one library members were successfully produced in recombinant form. In vitro assays employing a synthetic substrate incorporating a diene and a dienophile revealed broad-ranging cycloaddition activity amongst these polypeptides. The hypothetical protein Cyc15 was found to catalyse an intramolecular cycloaddition to generate a novel spirotetronate. The crystal structure of this enzyme, along with docking studies, establishes the basis for stereoselectivity in Cyc15, as compared to other spirotetronate cyclases.
Author(s): Zorn K, Back CR, Barringer R, Chadimova V, Manzo-Ruiz M, Mbatha SZ, Mobarec J-C, Williams SE, van der Kamp MW, Race PR, Willis CL, Hayes MA
Publication type: Article
Publication status: Published
Journal: ChemBioChem
Year: 2023
Volume: 24
Issue: 14
Print publication date: 17/07/2023
Online publication date: 12/06/2023
Acceptance date: 12/06/2023
Date deposited: 19/12/2023
ISSN (print): 1439-4227
ISSN (electronic): 1439-7633
Publisher: John Wiley and Sons Inc
URL: https://doi.org/10.1002/cbic.202300382
DOI: 10.1002/cbic.202300382
PubMed id: 37305956
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