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Inverse stable isotope labeling (InverSIL) links predicted catecholate siderophore gene clusters to their products in
Jose Miguel D Robes1,2, Tashi C E Liebergesell1,2, Victoria P Medvedeva1,2
1Department of Chemistry, University of Utah, Salt Lake City, Utah, USA.
Biorxiv : the Preprint Server for Biology
|November 24, 2025
Summary
This study introduces inverse stable isotope labeling (InverSIL) to identify bacterial siderophores and their biosynthetic gene clusters (BGCs). InverSIL successfully characterized known and discovered new siderophores, advancing our understanding of bacterial iron acquisition.
Area of Science:
- Microbiology
- Biochemistry
- Genomics
Background:
- Bacteria utilize siderophores for iron acquisition, but many predicted biosynthetic gene clusters (BGCs) lack characterized products.
- Determining siderophore structures is crucial for understanding bacterial iron metabolism and community interactions.
Purpose of the Study:
- To develop and apply a novel method, inverse stable isotope labeling (InverSIL), for rapid characterization of siderophore biosynthetic gene clusters (BGCs) and their products.
- To elucidate siderophore structures from various bacterial genera, including previously uncharacterized compounds.
Main Methods:
- Inverse stable isotope labeling (InverSIL) was employed, involving growth on 13C-labeled media and feeding unlabeled precursors.
- Mass spectrometry was used to identify and characterize siderophore products linked to specific BGCs.
- The method was applied to methylotrophic genera, Chromobacterium violaceum, Kushneria, Paracoccus, and Cellulomonas.
Main Results:
- InverSIL successfully linked predicted BGCs to their siderophore products.
- Structures of siderophores from Methylophilus, Methylobacterium, and Chromobacterium violaceum were determined.
- Unexpected enterobactin production was identified in Kushneria and Paracoccus.
- New siderophores, cellulochelin A and B, were discovered from Cellulomonas.
Conclusions:
- InverSIL is a powerful tool for functional BGC characterization and siderophore discovery.
- This work expands the known repertoire of bacterial siderophores and enhances understanding of iron acquisition mechanisms.
- The findings have implications for microbial ecology and the study of bacterial virulence.
