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Structure and Function of the α‐Hydroxylation Bimodule of the Mupirocin Polyketide Synthase

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Titel: Structure and Function of the α‐Hydroxylation Bimodule of the Mupirocin Polyketide Synthase
Autoren: Ashley J. Winter, R. Nisha Khanizeman, Abigail M. C. Barker‐Mountford, Andrew J. Devine, Luoyi Wang, Zhongshu Song, Jonathan A. Davies, Paul R. Race, Christopher Williams, Thomas J. Simpson, Christine L. Willis, Matthew P. Crump
Quelle: Angew Chem Weinheim Bergstr Ger
Angew Chem Int Ed Engl
Verlagsinformationen: Wiley, 2023.
Publikationsjahr: 2023
Schlagwörter: Polyketide Synthases/metabolism, Mupirocin, Forschungsartikel, Anti-Bacterial Agents/chemistry, Hydroxylation, Polyketide Synthases, Research Articles, Anti-Bacterial Agents
Beschreibung: 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.
Publikationsart: Article
Other literature type
Dateibeschreibung: application/pdf
Sprache: English
ISSN: 1521-3773
1433-7851
DOI: 10.1002/anie.202312514
DOI: 10.1002/ange.202312514
Zugangs-URL: https://pubmed.ncbi.nlm.nih.gov/37768840
https://pubmed.ncbi.nlm.nih.gov/38515435
Rights: CC BY
Dokumentencode: edsair.doi.dedup.....6c18bf6b4d4517a9868bbd79b975d4f1
Datenbank: OpenAIRE
Beschreibung
Abstract: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.
ISSN:15213773
14337851
DOI:10.1002/anie.202312514