Unlocking Cryptic Metabolites with Mass Spectrometry-Guided Transposon Mutant Selection.

TitleUnlocking Cryptic Metabolites with Mass Spectrometry-Guided Transposon Mutant Selection.
Publication TypeJournal Article
Year of Publication2020
AuthorsYoshimura, A, Covington, BC, Gallant, É, Zhang, C, Li, A, Seyedsayamdost, MR
JournalACS Chem Biol
Volume15
Issue10
Pagination2766-2774
Date Published2020 10 16
ISSN1554-8937
KeywordsBiological Products, Burkholderia, DNA Transposable Elements, Mass Spectrometry, Metabolome, Metabolomics, Multigene Family, Mutagenesis, Secondary Metabolism
Abstract

<p>The products of most secondary metabolite biosynthetic gene clusters (BGCs) have yet to be discovered, in part due to low expression levels in laboratory cultures. Reporter-guided mutant selection (RGMS) has recently been developed for this purpose: a mutant library is generated and screened, using genetic reporters to a chosen BGC, to select transcriptionally active mutants that then enable the characterization of the "cryptic" metabolite. The requirement for genetic reporters limits the approach to a single pathway within genetically tractable microorganisms. Herein, we utilize untargeted metabolomics in conjunction with transposon mutagenesis to provide a global read-out of secondary metabolism across large numbers of mutants. We employ self-organizing map analytics and imaging mass spectrometry to identify and characterize seven cryptic metabolites from mutant libraries of two different species. Applications of the methodologies reported can expand our understanding of the products and regulation of cryptic BGCs across phylogenetically diverse bacteria.</p>

DOI10.1021/acschembio.0c00558
Alternate JournalACS Chem Biol
PubMed ID32808751
PubMed Central IDPMC7572862
Grant ListDP2 AI124786 / AI / NIAID NIH HHS / United States