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Updated: May 16, 2026

Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues
Published on: November 22, 2014
Enzyme-directed mutasynthesis: a combined experimental and theoretical approach to substrate recognition of a
Uschi Sundermann1, Kenny Bravo-Rodriguez, Stephan Klopries
1Fakultät für Chemie, Chemische Biologie, Technische Universität Dortmund , Otto-Hahn-Str. 6, 44221 Dortmund, Germany.
Abstract:
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.
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