Dissecting modular synthases through inhibition: A complementary chemical and genetic approach

Christopher R. Vickery, Ian P. McCulloch, Eva Sonnenschein, Joris Beld, Joseph P. Noel, Michael D. Burkart*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Modular synthases, such as fatty acid, polyketide, and non-ribosomal peptide synthases (NRPSs), are sophisticated machineries essential in both primary and secondary metabolism. Various techniques have been developed to understand their genetic background and enzymatic abilities. However, uncovering the actual biosynthetic pathways remains challenging. Herein, we demonstrate a pipeline to study an assembly line synthase by interrogating the enzymatic function of each individual enzymatic domain of BpsA, a NRPS that produces the blue 3,3′-bipyridyl pigment indigoidine. Specific inhibitors for each biosynthetic domain of BpsA were obtained or synthesized, and the enzymatic performance of BpsA upon addition of each inhibitor was monitored by pigment development in vitro and in living bacteria. The results were verified using genetic mutants to inactivate each domain. Finally, the results complemented the currently proposed biosynthetic pathway of BpsA.
Original languageEnglish
Article number126820
JournalBioorganic & Medicinal Chemistry Letters
Volume30
Issue number2
Number of pages6
ISSN0960-894X
DOIs
Publication statusPublished - 2020

Keywords

  • Non-ribosomal peptide synthetase
  • Indigoidine
  • BpsA
  • Natural products
  • Domain-specific inhibitors

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