Methyltransferase Contingencies in the Pathway of Everninomicin D Antibiotics and Analogues. (7th September 2020)

• Record Type: Journal Article
• Authors: Limbrick, Emilianne M.; Yñigez‐Gutierrez, Audrey E.; Dulin, Callie C.; Derewacz, Dagmara K.; Spraggins, Jeffrey M.; McCulloch, Kathryn M.; Iverson, T. M.; Bachmann, Brian O.
• Is Part Of: Chembiochem. Volume 21:Number 23(2020)
• Page Start: 3349
• Page End: 3358
• Publication Date: 2020-09-07
• Subjects: actinomycetes -- antibiotic resistance -- biosynthesis -- everninomicin -- methyltransferases -- orthoesters; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.202000305
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: Everninomicins are orthoester oligosaccharide antibiotics with potent activity against multidrug‐resistant bacterial pathogens. Everninomicins act by disrupting ribosomal assembly in a distinct region in comparison to clinically prescribed drugs. We employed microporous intergeneric conjugation with Escherichia coli to manipulate Micromonospora for targeted gene‐replacement studies of multiple putative methyltransferases across the octasaccharide scaffold of everninomicin effecting the A1, C, F, and H rings. Analyses of gene‐replacement and genetic complementation mutants established the mutability of the everninomicin scaffold through the generation of 12 previously unreported analogues and, together with previous results, permitted assignment of the ten methyltransferases required for everninomicin biosynthesis. The in vitro activity of A1 ‐ and H‐ring‐modifying methyltransferases demonstrated the ability to catalyze late‐stage modification of the scaffold on an A1 ‐ring phenol and H‐ring C‐4' hydroxy moiety. Together these results establish the potential of the everninomicin scaffold for modification through mutagenesis and in vitro modification of advanced biosynthetic intermediates. Abstract : M. carbonacea species produce the everninomicin antibiotics, elaborate oligosaccharides with potent antibacterial activity. Herein, we demonstrate a genetic system for genome editing of everninomicin producers, including a new microporous intergeneric conjugation system for conjugal transfer, and analyze a host of methyltransferases spanning the octasaccharide scaffold. In the process, several new everninomicin analogues were generated.