Enzyme-Directed Mutasynthesis: A Combined Experimental and Theoretical Approach to Substrate Recognition of a Polyketide Synthase

Citations Over TimeTop 10% of 2012 papers

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.

Related Papers

• → Expanding Extender Substrate Selection for Unnatural Polyketide Biosynthesis by Acyltransferase Domain Exchange within a Modular Polyketide Synthase(2023)31 cited
• → A Close Look at a Ketosynthase from a Trans-Acyltransferase Modular Polyketide Synthase(2014)81 cited
• → Characterization of three succinyl-CoA acyltransferases involved in polyketide chain assembly(2023)2 cited
• → Recognition of (2 S )-Aminomalonyl-Acyl Carrier Protein (ACP) and (2 R )-Hydroxymalonyl-ACP by Acyltransferases in Zwittermicin A Biosynthesis(2010)33 cited
• → Biochemical Characterization of a Malonyl-Specific Acyltransferase Domain of FK506 Biosynthetic Polyketide Synthase(2014)10 cited