Supplementary Figure 6: Ferrioxamine family of metabolites is upregulated in WAC6273 engineered strains.
(A) Metabolic networking of WAC6273 n-butanolic extracts clustered a family of masses that were identified as ferrioxamines using the GNPS LC–MS/MS based dereplication platform18. Gray edges are shaded from light to dark scaled to the degree of relatedness (cosine score) between nodes. Each node is represented as a pie chart showing the relative abundance of each mass in wild-type WAC6273 (red) or streptothricin knockout Δorf15 pCRISPR-Cas9 (blue). Almost every member of the network is upregulated in the inactivated strain compared to wild-type, with numerous species being only detectable in the former. Pie charts are drawn based on one biological replicate but are representative of data collected from three biological replicates. (B) Structure of ferrioxamine family members. Numbering system for different analogs is based on Sidebottom et al19. Calculated and observed mass of [M+H]+ species are reported for high-resolution LC/MS data. (C) High-resolution LC–MS/MS was used to confirm network members as ferrioxamines. Calculated masses based on previously reported fragmentation patterns19 match observed masses in WAC6273 extracts. Fragmentation of a representative member, ferrioxamine 13, is shown. Fragmentation results are representative of two independent analyses.
