Defining the genes for the final steps in biosynthesis of the complex polyketide antibiotic mupirocin by Pseudomonas fluorescens NCIMB10586

Jack A Connolly, Amber Wilson, Malgorzata Macioszek, Zhongshu Song, Luoyi Wang, Hadi H Mohammad, Mukul Yadav, Maura di Martino, Claire E Miller, Joanne Hothersall, Anthony S Haines, Elton R Stephens, Matthew P Crump, Christine L Willis, Thomas J Simpson, Peter J Winn, Christopher M Thomas*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)

2 Citations (Scopus)
223 Downloads (Pure)

Abstract

The mupirocin trans-AT polyketide synthase pathway, provides a model system for manipulation of antibiotic biosynthesis. Its final phase involves removal of the tertiary hydroxyl group from pseudomonic acid B, PA-B, producing the fully active PA-A in a complex series of steps. To further clarify requirements for this conversion, we fed extracts containing PA-B to mutants of the producer strain singly deficient in each mup gene. This additionally identified mupM and mupN as required plus the sequence but not enzymic activity of mupL and ruled out need for other mup genes. A plasmid expressing mupLMNOPVCFU + macpE together with a derivative of the producer P. fluorescens strain NCIMB10586 lacking the mup cluster allowed conversion of PA-B to PA-A. MupN converts apo-mAcpE to holo-form while MupM is a mupirocin-resistant isoleucyl tRNA synthase, preventing self-poisoning. Surprisingly, the expression plasmid failed to allow the closely related P. fluorescens strain SBW25 to convert PA-B to PA-A.

Original languageEnglish
Article number1542
Number of pages9
JournalScientific Reports
Volume9
DOIs
Publication statusPublished - 7 Feb 2019

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    Connolly, J. A., Wilson, A., Macioszek, M., Song, Z., Wang, L., Mohammad, H. H., Yadav, M., di Martino, M., Miller, C. E., Hothersall, J., Haines, A. S., Stephens, E. R., Crump, M. P., Willis, C. L., Simpson, T. J., Winn, P. J., & Thomas, C. M. (2019). Defining the genes for the final steps in biosynthesis of the complex polyketide antibiotic mupirocin by Pseudomonas fluorescens NCIMB10586. Scientific Reports, 9, [1542]. https://doi.org/10.1038/s41598-018-38038-9