Novel RiPP Cyclodehydratases from Thermophilic Bacteria Edit Folded Polypeptides
Year: 2026
Type: preprint
Abstract: Ribosomally synthesized and post-translationally modified peptides (RiPPs) often contain backbone-embedded oxazoline/thiazoline heterocycles that are installed by enzymes known as cyclodehydratases. These editing enzymes are of great interest as biocatalysts for biotechnology applications, as embedded oxazoline/thiazoline heterocycles can induce non-subtle structural and functional changes that alter bioactivity both directly and indirectly. However, the toolkit of existing cyclodehydratases with robust activity, stability, and well-defined sequence specificity is extremely limited. Herein, we apply a genome mining approach to discover two novel cyclodehydratases with improved thermostability: TcuD (from Thermomonospora curvata ) and TteD (from Thermocatellispora tengchongensis ). Notably, TcuD also displays exceptional substrate scope. Specifically, TcuD showed remarkable tolerance to mutations, insertions, and deletions to its substrate precursor peptide, TcuA , and it easily modified a diverse set of substrates whose sequences were derived from α-helical regions of structurally characterized proteins. This work expands the repertoire of cyclodehydratase enzymes available for polypeptide editing applications in biotechnology.
Source: ChemRxiv
Institutions: University of California System, University of New Mexico
Language: English
Subfield: Applied Microbiology and Biotechnology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7204173728