Sundermann, Uschi Bravo-Rodriguez, Kenny Klopries, Stephan Kushnir, Susanna Gomez, Hansel Sanchez-Garcia, Elsa Schulz, Frank Enzyme-Directed Mutasynthesis: A Combined Experimental and Theoretical Approach to Substrate Recognition of a Polyketide Synthase Acyltransferase 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. substrate recognition;Polyketide SynthaseAcyltransferase domains control;PKS domains;Polyketide Synthases;biosynthesi;product derivatives;Theoretical Approach;malonic acid thioester;Substrate Recognition;product analogues;extender unit recognition;incorporation;Experimental;acyltransferase substrate profile;malonic acid building block;enzyme engineering strategy;point mutation 2016-02-19
    https://acs.figshare.com/articles/journal_contribution/Enzyme_Directed_Mutasynthesis_A_Combined_Experimental_and_Theoretical_Approach_to_Substrate_Recognition_of_a_Polyketide_Synthase/2442655
10.1021/cb300505w.s001