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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. Angewandte Chemie International Edition published by Wiley-VCH GmbH.Mupirocin is a clinically important antibiotic produced by a trans-AT Type I polyketide synthase (PKS) in Pseudomonas fluorescens. The major bioactive metabolite, pseudomonic acid A (PA−A), is assembled on a tetrasubstituted tetrahydropyran (THP) core incorporating a 6-hydroxy group proposed to be introduced by α-hydroxylation of the thioester of the acyl carrier protein (ACP) bound polyketide chain. Herein, we describe an in vitro approach combining purified enzyme components, chemical synthesis, isotopic labelling, mass spectrometry and NMR in conjunction with in vivo studies leading to the first characterisation of the α-hydroxylation bimodule of the mupirocin biosynthetic pathway. These studies reveal the precise timing of hydroxylation by MupA, substrate specificity and the ACP dependency of the enzyme components that comprise this α-hydroxylation bimodule. Furthermore, using purified enzyme, it is shown that the MmpA KS0 shows relaxed substrate specificity, suggesting precise spatiotemporal control of in trans MupA recruitment in the context of the PKS. Finally, the detection of multiple intermodular MupA/ACP interactions suggests these bimodules may integrate MupA into their assembly.
Author(s): Winter AJ, Khanizeman RN, Barker-Mountford AMC, Devine AJ, Wang L, Song Z, Davies JA, Race PR, Williams C, Simpson TJ, Willis CL, Crump MP
Publication type: Article
Publication status: Published
Journal: Angewandte Chemie International Edition
Year: 2023
Volume: 62
Issue: 47
Print publication date: 20/11/2023
Online publication date: 28/09/2023
Acceptance date: 28/09/2023
Date deposited: 19/12/2023
ISSN (print): 1433-7851
ISSN (electronic): 1521-3773
Publisher: John Wiley and Sons Inc
URL: https://doi.org/10.1002/anie.202312514
DOI: 10.1002/anie.202312514
PubMed id: 37768840
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