ePrints

Interrogation of an Enzyme Library Reveals the Catalytic Plasticity of Naturally Evolved [4+2] Cyclases

Lookup NU author(s): Professor Paul RaceORCiD

Downloads

• Published version [.pdf]

Licence

This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

© 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.

Publication metadata

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

Data Access Statement: The data that support the findings of this study are available from the corresponding author upon reasonable request.

PubMed id: 37305956

Altmetrics

Funding

Share