Mass Spectrometry guided discovery and design of novel Asperphenamate analogues from Penicilium astrolabium reveals an extraordinary NRPS flexibility

Karolina Subko, Xinhui Wang, Frederik H. Nielsen, Thomas Isbrandt Petersen, Charlotte H. Gotfredsen, Carmen Ramos, Thomas Mackenzie, Francisca Vicente, Olga Genilloud, Jens C. Frisvad, Thomas O. Larsen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Asperphenamate is a small peptide natural product that has gained much interest due to its antitumor activity. In the recent years numerous bioactive synthetic asperphenamate analogues have been reported, whereas only a handful of natural analogues either of microbial or plant origin has been discovered. Herein we describe a UHPLC-HRMS/MS and amino acid supplement approach for discovery and design novel asperphenamate analogues. Chemical analysis of Penicillium astrolabium, a prolific producer of asperphenamate, revealed three previously described and two novel asperphenamate analogues produced in significant amounts, suggesting a potential for biosynthesis of further asperphenamate analogues by varying the amino acid availability. Subsequent growth on proteogenic and non-proteogenic amino acid enriched media, revealed a series of novel asperphenamate analogues, including single or double amino acid exchange, as well as benzoic acid exchange for nicotinic acid, with the latter observed from a natural source for the first time. In total, 22 new asperphenamate analogues were characterized by HRMS/MS, with one additionally confirmed by isolation and NMR structure elucidation. This study indicates an extraordinary nonribosomal peptide synthetase (NRPS) flexibility based on substrate availability, and therefore the potential for manipulating and designing novel peptide natural products in filamentous fungi.
Original languageEnglish
Article number618730
JournalFrontiers in Microbiology
Volume11
Number of pages13
ISSN1664-302X
DOIs
Publication statusPublished - 2021

Keywords

  • Natural product discovery
  • Mass spectrometry
  • Filamentous fungi
  • Asperphenamate
  • Amino acid incorporation
  • Biological activity
  • NRPS flexibility

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