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Navigating the conjugated metabolome.

Shipei Xing1,2, Abubaker Patan1,2, Julius Agongo1,2

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This study maps the vast, underexplored world of conjugated metabolites using billions of spectra. It reveals thousands of new structures, expanding our understanding of biochemical diversity and potential bioactivities.

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Area of Science:

  • Biochemistry
  • Metabolomics
  • Chemical Biology

Background:

  • Metabolomics has cataloged many small molecules, but conjugated metabolites remain largely unmapped.
  • Conjugated metabolites play roles as microbial signals, detoxification intermediates, and endogenous regulators.

Purpose of the Study:

  • To map conjugation events and discover novel conjugated metabolites across public metabolomics data.
  • To establish a resource for structural discovery and MS/MS annotation of conjugated metabolites.

Main Methods:

  • Mined 1.32 billion MS/MS spectra from public repositories using reverse spectral searching and delta-mass inference.
  • Generated structural hypotheses for millions of MS/MS clusters and inferred substructure pairs and candidate conjugates.

Main Results:

  • Identified over 217,000 substructure pairs and 3.4 million candidate conjugates.
  • Predicted conjugates include host-microbe co-metabolites, diet-derived, and drug-derived species, such as drug-ethanolamine and steroid-phosphoethanolamine conjugates.
  • Validated 55 conjugates using synthetic standards and 27 with retention time matching in biological samples.

Conclusions:

  • Delivered a comprehensive map of potential conjugation chemistry across multiple repositories.
  • Established a valuable resource for exploring the conjugated metabolome's scope and diversity.
  • Provided a scalable framework for future structural discovery and MS/MS annotation in metabolomics.