Genome Mining Reveals trans‐AT Polyketide Synthase Directed Antibiotic Biosynthesis in the Bacterial Phylum Bacteroidetes

ChemBioChem
2010.0

Abstract

Complex polyketides such as erythromycin, epothilone, and FK506 greatly contribute to human health and are synthesized by modular polyketide synthases (PKSs). A novel, evolutionarily distinct group of trans-AT PKSs—using free-standing acyltransferases (AT) instead of integrated AT domains—has been linked to diverse bioactive polyketides but was long overlooked. Building on a predictive approach for trans-AT PKSs based on ketosynthase (KS) domain phylogeny (correlating KS position in phylogenetic trees with substrate specificity), we applied trans-AT genome mining to Chitinophaga pinensis DSM 2588, a member of the chemically poorly studied Bacteroidetes phylum. We identified a 75.1 kb trans-AT PKS system (els PKS) with 18 genes: six multimodular PKS proteins lack integrated AT domains, while free-standing ATs (ElsA, ElsB) are present (ElsA also harbors a C-terminal oxidoreductase domain for trans enoyl reduction). To predict the polyketide structure, a phylogenetic tree was constructed from 413 KS sequences (catalyzing polyketide chain extensions). Most els KSs fell into functional clades with known substrate specificities, enabling high-confidence deduction of the polyketide structure despite aberrant PKS features (e.g., split modules, mutated HGTGT motifs essential for elongation). A conserved sequence motif (CGVYLGIMS/N/GN/SE) specific to a clade with β-methyl groups on E double bonds was also identified, facilitating rapid function prediction. Our study links the els PKS to the biosynthesis of elansolids—antibacterial and cytotoxic agents with unusual post-PKS modifications—extends trans-AT genome mining to the Bacteroidetes phylum, and validates the predictive power of KS phylogenetic analysis for non-canonical PKS pathways.

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