Title | Unlocking Cryptic Metabolites with Mass Spectrometry-Guided Transposon Mutant Selection. |
Publication Type | Journal Article |
Year of Publication | 2020 |
Authors | Yoshimura, A, Covington, BC, Gallant, É, Zhang, C, Li, A, Seyedsayamdost, MR |
Journal | ACS Chem Biol |
Volume | 15 |
Issue | 10 |
Pagination | 2766-2774 |
Date Published | 2020 Oct 16 |
ISSN | 1554-8937 |
Keywords | Biological 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> |
DOI | 10.1021/acschembio.0c00558 |
Alternate Journal | ACS Chem Biol |
PubMed ID | 32808751 |
PubMed Central ID | PMC7572862 |
Grant List | DP2 AI124786 / AI / NIAID NIH HHS / United States |