Acyl‐Chain Elongation Drives Ketosynthase Substrate Selectivity in trans‐Acyltransferase Polyketide Synthases1. (21st December 2014)

• Record Type: Journal Article
• Authors: Jenner, Matthew; Afonso, José Pedro; Bailey, Hannah R.; Frank, Sarah; Kampa, Annette; Piel, Jörn; Oldham, Neil J.
• Is Part Of: Angewandte Chemie. Volume 127:Number 6(2015)
• Page Start: 1837
• Page End: 1841
• Publication Date: 2014-12-21
• Subjects: Biosynthese -- Ketosynthasen -- Massenspektrometrie -- Polyketide -- Polyketid‐Synthasen; Chemistry -- Periodicals; 540
• DOI: 10.1002/ange.201410219
• Languages: English
• ISSNs: 0044-8249

Abstract

Abstract: Type I modular polyketide synthases (PKSs), which are responsible for the biosynthesis of many biologically active agents, possess a ketosynthase (KS) domain within each module to catalyze chain elongation. Acylation of the KS active site Cys residue is followed by transfer to malonyl‐ACP to yield an extended β‐ketoacyl chain (ACP=acyl carrier protein). To date, the precise contribution of KS selectivity in controlling product fidelity has been unclear. Six KS domains from trans ‐acyltransferase (trans‐AT) PKSs were subjected to a mass spectrometry based elongation assay, and higher substrate selectivity was identified for the elongating step than in preceding acylation. A close correspondence between the observed KS selectivity and that predicted by phylogenetic analysis was seen. These findings provide insights into the mechanism of KS selectivity in this important group of PKSs, can serve as guidance for engineering, and show that targeted mutagenesis can be used to expand the repertoire of acceptable substrates.