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Rational engineering of a thermostable α-oxoamine synthase biocatalyst expands the substrate scope and synthetic applicability

Lookup NU author(s): Dr Arnaud Basle, Dr Jon Marles-WrightORCiD

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

Abstract

© The Author(s) 2025.Carbon-carbon bond formation is one of the key pillars of organic synthesis. Green, selective and efficient biocatalytic methods for such are therefore highly desirable. The α-oxoamine synthases (AOSs) are a class of pyridoxal 5’-phosphate (PLP)-dependent, irreversible, carbon-carbon bond-forming enzymes, which have been limited previously by their narrow substrate specificity and requirement of acyl-CoA thioester substrates. We recently characterized a thermophilic enzyme from Thermus thermophilus (ThAOS) with a much broader substrate scope and described its use in a chemo-biocatalytic cascade process to generate pyrroles in good yields and timescales. Herein, we report the structure-guided engineering of ThAOS to arrive at variants able to use a greatly expanded range of amino acid and simplified N-acetylcysteamine (SNAc) acyl-thioester substrates. The crystal structure of the improved ThAOS V79A variant with a bound PLP:l-penicillamine external aldimine ligand, provides insight into the properties of the engineered biocatalyst.

Publication metadata

Author(s): Ashley B, Mathew S, Sajjad M, Zhu Y, Novikovs N, Basle A, Marles-Wright J, Campopiano DJ

Publication type: Article

Publication status: Published

Journal: Communications Chemistry

Year: 2025

Volume: 8

Issue: 1

Online publication date: 13/03/2025

Acceptance date: 05/02/2025

Date deposited: 08/04/2025

ISSN (electronic): 2399-3669

Publisher: Nature Research

URL: https://doi.org/10.1038/s42004-025-01448-8

DOI: 10.1038/s42004-025-01448-8

Data Access Statement: Protein structure raw data files (MTZ and PDB for 8S1Y) are available from the author. Raw data files for HPLC, NMR, kinetic and UV-vis data are also available upon request.

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