Sequential In Vitro Cyclization by Cytochrome P450 Enzymes of Glycopeptide Antibiotic Precursors Bearing the X‐Domain from Nonribosomal Peptide Biosynthesis. (9th November 2015)

• Record Type: Journal Article
• Authors: Brieke, Clara; Peschke, Madeleine; Haslinger, Kristina; Cryle, Max J.
• Is Part Of: Angewandte Chemie. Volume 127:Number 52(2015)
• Page Start: 15941
• Page End: 15945
• Publication Date: 2015-11-09
• Subjects: Biosynthese -- Glykopeptid‐Antibiotika -- Nichtribosomale Peptidsynthetasen -- P450‐Cytochrome -- Peptidvernetzung; Chemistry -- Periodicals; 540
• DOI: 10.1002/ange.201507533
• Languages: English
• ISSNs: 0044-8249

Abstract

Abstract: The biosynthesis of the glycopeptide antibiotics, which include vancomycin and teicoplanin, relies on the interplay between the peptide‐producing non‐ribosomal peptide synthetase (NRPS) and Cytochrome P450 enzymes (P450s) that catalyze side‐chain crosslinking of the peptide. We demonstrate that sequential in vitro P450‐catalyzed cyclization of peptide substrates is enabled by the use of an NRPS peptide carrier protein (PCP)‐X di‐domain as a P450 recruitment platform. This study reveals that whilst the precursor peptide sequence influences the installation of the second crosslink by the P450 OxyAtei, activity is not restricted to the native teicoplanin peptide. Initial peptide cyclization is possible with teicoplanin and vancomycin OxyB homologues, and the latter displays excellent activity with all substrate combinations tested. By using non‐natural X‐domain substrates, bicyclization of hexapeptides was also shown, which demonstrates the utility of this method for the cyclization of varied peptide substrates in vitro. Abstract : Interne Vernetzung : Der zweifache Ringschluss von Vorläuferpeptiden für Glykopeptid‐Antibiotika konjugiert an Proteine, die sich von nichtribosomalen Peptidsynthetasen (NRPS) ableiten und die X‐Domäne enthalten, wird durch die beiden Cytochrom‐P450‐Enzyme OxyB und OxyA katalysiert. Die P450‐Enzyme erweisen sich dabei als bemerkenswert tolerant bezüglich alternativer Substratstrukturen.