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

• Published in: Scientific reports 2019-02, Vol.9 (1), p.1542, Article 1542
• Main Authors: Connolly, Jack A., Wilson, Amber, Macioszek, Malgorzata, Song, Zhongshu, Wang, Luoyi, Mohammad, Hadi H., Yadav, Mukul, di Martino, Maura, Miller, Claire E., Hothersall, Joanne, Haines, Anthony S., Stephens, Elton R., Crump, Matthew P., Willis, Christine L., Simpson, Thomas J., Winn, Peter J., Thomas, Christopher M.
• Format: Article
• Language: English
• Subjects: 45; 45/41; 45/44; 45/70; 631/326/22/1290; 631/92/349/977; Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - metabolism; Antibiotics; Bacillus subtilis - genetics; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biosynthesis; Escherichia coli - genetics; Humanities and Social Sciences; multidisciplinary; Mupirocin; Mupirocin - biosynthesis; Mupirocin - chemistry; Mutagenesis; Plasmids - genetics; Plasmids - metabolism; Polyketide synthase; Polyketide Synthases - genetics; Polyketide Synthases - metabolism; Polyketides - chemistry; Polyketides - metabolism; Pseudomonas fluorescens - genetics; Pseudomonas fluorescens - metabolism; Science; Science (multidisciplinary); tRNA
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-018-38038-9

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.