TY - JOUR
T1 - Unlocking Cryptic Metabolites with Mass Spectrometry-Guided Transposon Mutant Selection
AU - Yoshimura, Aya
AU - Covington, Brett C.
AU - Gallant, Étienne
AU - Zhang, Chen
AU - Li, Anran
AU - Seyedsayamdost, Mohammad R.
N1 - Publisher Copyright:
Copyright © 2020 American Chemical Society.
PY - 2020/10/16
Y1 - 2020/10/16
N2 - 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 Burkholderia species. Applications of the methodologies reported can expand our understanding of the products and regulation of cryptic BGCs across phylogenetically diverse bacteria.
AB - 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 Burkholderia species. Applications of the methodologies reported can expand our understanding of the products and regulation of cryptic BGCs across phylogenetically diverse bacteria.
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U2 - 10.1021/acschembio.0c00558
DO - 10.1021/acschembio.0c00558
M3 - Article
C2 - 32808751
AN - SCOPUS:85093538846
SN - 1554-8929
VL - 15
SP - 2766
EP - 2774
JO - ACS chemical biology
JF - ACS chemical biology
IS - 10
ER -