Programmed Iteration Controls the Assembly of the Nonanoic Acid Side Chain of the Antibiotic Mupirocin

Ashley James Winter; Matthew T Rowe; Angus N M Weir; Nahida Akter; Sbusisiwe Z Mbatha; Paul D Walker; Christopher Williams; Zhongshu Song; Paul R Race; Christine L Willis; Matthew Philip Crump

doi:10.1002/anie.202212393

Programmed Iteration Controls the Assembly of the Nonanoic Acid Side Chain of the Antibiotic Mupirocin

Ashley James Winter, Matthew T Rowe, Angus N M Weir, Nahida Akter, Sbusisiwe Z Mbatha, Paul D Walker, Christopher Williams, Zhongshu Song, Paul R Race, Christine L Willis^*, Matthew Philip Crump^*

^*Corresponding author for this work

Research output: Contribution to journal › Article (Academic Journal) › peer-review

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Abstract

Mupirocin is a clinically important antibiotic produced by Pseudomonas fluorescens NCIMB 10586 that is assembled by a complex trans-AT polyketide synthase. The polyketide fragment, monic acid, is esterified by a 9-hydroxynonanoic acid (9HN) side-chain which is essential for biological activity. The ester side-chain assembly is initialised from a 3-hydroxypropionate (3HP) starter unit attached to the acyl carrier protein (ACP) MacpD, but the fate of this species is unknown. Here we report the application of NMR, mass spectrometry, chemical probes and in vitro assays to establish the remaining steps of 9HN biosynthesis. These investigations reveal a complex interplay between a novel iterative or 'stuttering' KS-AT didomain (MmpF), the multi-domain module MmpB and multiple ACPs. This work has important implications for understanding the late-stage biosynthetic steps of mupirocin and will be important for future engineering of related trans-AT biosynthetic pathways (eg thiomarinol).

Original language	English
Article number	e202212393
Journal	Angewandte Chemie - International Edition
Volume	61
Issue number	50
Early online date	13 Oct 2022
DOIs	https://doi.org/10.1002/anie.202212393
Publication status	Published - 5 Dec 2022

Bibliographical note

Funding Information:
We thank the BBSRC for funding (BB/R007853/1 and BB/W008823/1, A. Winter and Z. Song) and BBSRC/EPSRC for funding through the Bristol Centre for Synthetic Biology (BB/L01386X/1), the Bristol Chemical Synthesis Centre for Doctoral Training for P. Walker (EP/L015366/1) and the doctoral training grant for A Weir. We also thank the British Commonwealth for funding for N. Akter (BDCS‐2017‐50) and the BBSRC SWBIO DTP2 (BB/M009122/1) (M. Rowe).

Publisher Copyright:
© 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.

Structured keywords

BCS and TECS CDTs
Bristol BioDesign Institute
BrisSynBio

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Winter, A. J., Rowe, M. T., Weir, A. N. M., Akter, N., Mbatha, S. Z., Walker, P. D., Williams, C., Song, Z., Race, P. R., Willis, C. L., & Crump, M. P. (2022). Programmed Iteration Controls the Assembly of the Nonanoic Acid Side Chain of the Antibiotic Mupirocin. Angewandte Chemie - International Edition, 61(50), Article e202212393. https://doi.org/10.1002/anie.202212393

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abstract = "Mupirocin is a clinically important antibiotic produced by Pseudomonas fluorescens NCIMB 10586 that is assembled by a complex trans-AT polyketide synthase. The polyketide fragment, monic acid, is esterified by a 9-hydroxynonanoic acid (9HN) side-chain which is essential for biological activity. The ester side-chain assembly is initialised from a 3-hydroxypropionate (3HP) starter unit attached to the acyl carrier protein (ACP) MacpD, but the fate of this species is unknown. Here we report the application of NMR, mass spectrometry, chemical probes and in vitro assays to establish the remaining steps of 9HN biosynthesis. These investigations reveal a complex interplay between a novel iterative or 'stuttering' KS-AT didomain (MmpF), the multi-domain module MmpB and multiple ACPs. This work has important implications for understanding the late-stage biosynthetic steps of mupirocin and will be important for future engineering of related trans-AT biosynthetic pathways (eg thiomarinol).",

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note = "Funding Information: We thank the BBSRC for funding (BB/R007853/1 and BB/W008823/1, A. Winter and Z. Song) and BBSRC/EPSRC for funding through the Bristol Centre for Synthetic Biology (BB/L01386X/1), the Bristol Chemical Synthesis Centre for Doctoral Training for P. Walker (EP/L015366/1) and the doctoral training grant for A Weir. We also thank the British Commonwealth for funding for N. Akter (BDCS‐2017‐50) and the BBSRC SWBIO DTP2 (BB/M009122/1) (M. Rowe). Publisher Copyright: {\textcopyright} 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.",

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AU - Mbatha, Sbusisiwe Z

AU - Walker, Paul D

AU - Williams, Christopher

AU - Song, Zhongshu

AU - Race, Paul R

AU - Willis, Christine L

AU - Crump, Matthew Philip

N1 - Funding Information: We thank the BBSRC for funding (BB/R007853/1 and BB/W008823/1, A. Winter and Z. Song) and BBSRC/EPSRC for funding through the Bristol Centre for Synthetic Biology (BB/L01386X/1), the Bristol Chemical Synthesis Centre for Doctoral Training for P. Walker (EP/L015366/1) and the doctoral training grant for A Weir. We also thank the British Commonwealth for funding for N. Akter (BDCS‐2017‐50) and the BBSRC SWBIO DTP2 (BB/M009122/1) (M. Rowe). Publisher Copyright: © 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.

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N2 - Mupirocin is a clinically important antibiotic produced by Pseudomonas fluorescens NCIMB 10586 that is assembled by a complex trans-AT polyketide synthase. The polyketide fragment, monic acid, is esterified by a 9-hydroxynonanoic acid (9HN) side-chain which is essential for biological activity. The ester side-chain assembly is initialised from a 3-hydroxypropionate (3HP) starter unit attached to the acyl carrier protein (ACP) MacpD, but the fate of this species is unknown. Here we report the application of NMR, mass spectrometry, chemical probes and in vitro assays to establish the remaining steps of 9HN biosynthesis. These investigations reveal a complex interplay between a novel iterative or 'stuttering' KS-AT didomain (MmpF), the multi-domain module MmpB and multiple ACPs. This work has important implications for understanding the late-stage biosynthetic steps of mupirocin and will be important for future engineering of related trans-AT biosynthetic pathways (eg thiomarinol).

AB - Mupirocin is a clinically important antibiotic produced by Pseudomonas fluorescens NCIMB 10586 that is assembled by a complex trans-AT polyketide synthase. The polyketide fragment, monic acid, is esterified by a 9-hydroxynonanoic acid (9HN) side-chain which is essential for biological activity. The ester side-chain assembly is initialised from a 3-hydroxypropionate (3HP) starter unit attached to the acyl carrier protein (ACP) MacpD, but the fate of this species is unknown. Here we report the application of NMR, mass spectrometry, chemical probes and in vitro assays to establish the remaining steps of 9HN biosynthesis. These investigations reveal a complex interplay between a novel iterative or 'stuttering' KS-AT didomain (MmpF), the multi-domain module MmpB and multiple ACPs. This work has important implications for understanding the late-stage biosynthetic steps of mupirocin and will be important for future engineering of related trans-AT biosynthetic pathways (eg thiomarinol).

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DO - 10.1002/anie.202212393

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ER -

Programmed Iteration Controls the Assembly of the Nonanoic Acid Side Chain of the Antibiotic Mupirocin

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