Thiazoline-specific amidohydrolase PurAH is the gatekeeper of bottromycin biosynthesis

Sikandar, A., Franz, L., Melse, O., Antes, I. and Koehnke, J. (2019) Thiazoline-specific amidohydrolase PurAH is the gatekeeper of bottromycin biosynthesis. Journal of the American Chemical Society, 141(25), pp. 9748-9752. (doi: 10.1021/jacs.8b12231) (PMID:31192589)

Abstract

The ribosomally synthesized and post-translationally modified peptide (RiPP) bottromycin A2 possesses potent antimicrobial activity. Its biosynthesis involves the enzymatic formation of a macroamidine, a process previously suggested to require the concerted efforts of a YcaO enzyme (PurCD) and an amidohydrolase (PurAH) in vivo. In vitro, PurCD alone is sufficient to catalyze formation of the macroamidine, but the process is reversible. We set out to probe the role of PurAH in macroamidine formation in vitro. We demonstrate that PurAH is highly selective for macroamidine-containing precursor peptides and cleaves C-terminal of a thiazoline, thus removing the follower peptide. After follower cleavage, macroamidine formation is irreversible, indicating PurAH as the gatekeeper of bottromycin biosynthesis. The structure of PurAH suggests residues involved in catalysis, which were probed through mutagenesis.

• Item Type: Articles
• Additional Information: J.K. thanks the German Research Foundation for an Emmy Noether Fellowship (KO 4116/3-1). O.M. and I.A. thank the German Research Foundation for financial support by the SFB 749, project C08 and CIPSM.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Koehnke, Professor Jesko
• Authors: Sikandar, A., Franz, L., Melse, O., Antes, I., and Koehnke, J.
• College/School: College of Science and Engineering > School of Chemistry
• Journal Name: Journal of the American Chemical Society
• Publisher: American Chemical Society
• ISSN: 0002-7863
• ISSN (Online): 1520-5126
• Published Online: 07 June 2019
• Copyright Holders: Copyright © 2019 American Chemical Society
• First Published: First published in Journal of the American Chemical Society 141(25):9748-9752
• Publisher Policy: Reproduced in accordance with the copyright policy of the publisher