In Vitro Biosynthesis of the Core Scaffold of the Thiopeptide Thiomuracin
Year: 2015
Type: article
Abstract: Thiopeptides are potent antibiotics that inhibit protein synthesis. They are made by a remarkable post-translational modification process that transforms a linear peptide into a polycyclic structure. We present here the in vitro biosynthesis of the core scaffold of thiomuracin catalyzed by six proteins. We show that cyclodehydration precedes dehydration, and that dehydration is catalyzed by two proteins in a tRNA(Glu)-dependent manner. The enzyme that generates the pyridine core from two dehydroalanines ejects the leader peptide as a C-terminal carboxamide. Mutagenesis studies of the enzyme TbtD identified important residues for a formal [4+2] cycloaddition process. The core structure of thiomuracin exhibits similar antimicrobial activity to other known congeners, illustrating that in vitro biosynthesis is a viable route to potent antibiotics that can be explored for the rapid and renewable generation of analogues.
DOI: 10.1021/jacs.5b10194
Language: English
Subfield: Pharmacology (medical)
Field: Medicine
Domain: Health Sciences
Open Access status: hybrid
Estimated APC paid: $4,500
Awards: G3723409455, 900MHz NMR for Structural Biology in Chicago, Enzyme Chemistry and Biological Function of RiPP-like Modifications., G5739928637, 4800 Plus MALDI TOF/TOF Analyzer, Posttranslational Modifications in Natural Product Biosynthesis, G7840311943, A common denominator of pathogenesis; a rare opportunity for novel therapeutic de
OpenAlex ID: W2231071237