Abstract
Erythromycin and related macrolide antibiotics are widely used polyketide natural products. We have evolved an engineered biosynthetic pathway in Escherichia coli that yields erythromycin analogs from simple synthetic precursors. Multiple rounds of mutagenesis and screening led to the identification of new mutant strains with improved efficiency for precursor-directed biosynthesis. Genetic and biochemical analysis suggested that the phenotypically relevant alterations in these mutant strains were localized exclusively to the host-vector system, and not to the polyketide synthase. We also demonstrate the utility of this improved system through engineered biosynthesis of a novel alkynyl erythromycin derivative with comparable antibacterial activity to its natural counterpart. In addition to reinforcing the power of directed evolution for engineering macrolide biosynthesis, our studies have identified a new lead substance for investigating structure–function relationships in the bacterial ribosome.
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Acknowledgements
This research was supported by grants from the National Institutes of Health to C.K. (GM 087934) and D.E.C. (GM 22172). C.J.B.H. is supported by a fellowship from the Stanford Center for Molecular Analysis and Design (CMAD).
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Dedicated to the late Dr C Richard Hutchinson for his exceptional contributions to natural product biosynthesis, engineering, and drug discovery.
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Lee, H., Harvey, C., Cane, D. et al. Improved precursor-directed biosynthesis in E. coli via directed evolution. J Antibiot 64, 59–64 (2011). https://doi.org/10.1038/ja.2010.129
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DOI: https://doi.org/10.1038/ja.2010.129
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