1. Evolution-guided engineering of trans -acyltransferase polyketide synthases.
- Author
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Mabesoone MFJ, Leopold-Messer S, Minas HA, Chepkirui C, Chawengrum P, Reiter S, Meoded RA, Wolf S, Genz F, Magnus N, Piechulla B, Walker AS, and Piel J
- Subjects
- Serratia, Amino Acid Motifs, Acyltransferases genetics, Acyltransferases chemistry, Polyketide Synthases chemistry, Polyketide Synthases genetics, Polyketides chemistry, Directed Molecular Evolution, Bacterial Proteins chemistry, Bacterial Proteins genetics, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins genetics
- Abstract
Bacterial multimodular polyketide synthases (PKSs) are giant enzymes that generate a wide range of therapeutically important but synthetically challenging natural products. Diversification of polyketide structures can be achieved by engineering these enzymes. However, notwithstanding successes made with textbook cis -acyltransferase ( cis -AT) PKSs, tailoring such large assembly lines remains challenging. Unlike textbook PKSs, trans -AT PKSs feature an extraordinary diversity of PKS modules and commonly evolve to form hybrid PKSs. In this study, we analyzed amino acid coevolution to identify a common module site that yields functional PKSs. We used this site to insert and delete diverse PKS parts and create 22 engineered trans -AT PKSs from various pathways and in two bacterial producers. The high success rates of our engineering approach highlight the broader applicability to generate complex designer polyketides.
- Published
- 2024
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