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Enzyme-Directed Mutasynthesis: A Combined Experimental and Theoretical Approach to Substrate Recognition of a Polyketide Synthase
journal contribution
posted on 2016-02-19, 22:30 authored by Uschi Sundermann, Kenny Bravo-Rodriguez, Stephan Klopries, Susanna Kushnir, Hansel Gomez, Elsa Sanchez-Garcia, Frank SchulzAcyltransferase domains control the extender unit recognition
in
Polyketide Synthases (PKS) and thereby the side-chain diversity of
the resulting natural products. The enzyme engineering strategy presented
here allows the alteration of the acyltransferase substrate profile
to enable an engineered biosynthesis of natural product derivatives
through the incorporation of a synthetic malonic acid thioester. Experimental
sequence–function correlations combined with computational
modeling revealed the origins of substrate recognition in these PKS
domains and enabled a targeted mutagenesis. We show how a single point
mutation was able to direct the incorporation of a malonic acid building
block with a non-native functional group into erythromycin. This approach,
introduced here as enzyme-directed mutasynthesis, opens a new field
of possibilities beyond the state of the art for the combination of
organic chemistry and biosynthesis toward natural product analogues.
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substrate recognitionPolyketide SynthaseAcyltransferase domains controlPKS domainsPolyketide Synthasesbiosynthesiproduct derivativesTheoretical Approachmalonic acid thioesterSubstrate Recognitionproduct analoguesextender unit recognitionincorporationExperimentalacyltransferase substrate profilemalonic acid building blockenzyme engineering strategypoint mutation
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