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Poster Presentations
Day 4, June 25（Wed.）
Room P (Maesato East, Foyer, Ocean Wing)
- 4P-AM-34
iprm-PASEF MALDI MS/MS Imaging Reveals Specialized Metabolites Produced by the Cheese Rind Microbiome
(1UCSC, 2BRUKER, 3BRUKERSG)
oWen Donq Looi3, Robert Shepard1, Gordon Luu2, Sumankalai Ramachandran2, Azad Eshghi2, Laura Sanchez1
Confident annotation in MALDI-imaging has historically been a significant challenge, stemming from the difficulty in deploying large-scale MS/MS in imaging applications. The recent introduction of imaging parallel reaction monitoring with parallel accumulation serial fragmentation (iprm-PASEF®), has enabled the acquisition of spatially resolved, high resolution MS/MS data for up to 25 mobility-resolved precursors. Here, we co-cultured Penicillium solitum #12 and Glutamicibacter arilaitensis JB182 on 2.5% rind cheese curd agar, subsequently spotted on a MALDI ground steel plate. A TIMS enabled MS1 imaging scan was performed on these spots. A modified T-ReX3 feature finding was utilized to generate a feature list from the MS1 imaging dataset, which was further analyzed by SCiLStm Lab 2025b to identify a list of ten potential precursors that could be produced under these co-culture conditions. Subsequently, MS/MS spectra were acquired by iprm-PASEF based on this precursor list. Bruker MetaboScape® 2025 was employed to analyze the MS/MS spectra and annotate these features. This annotation workflow successfully identified one precursor of interest (m/z 655.2733 ± 0.05 Da, 1/K0 1.2729 ± 0.02) as a porphyrin class compound, consistent with prior studies showing that Glutamicibacter arilaitensis JB182 produces the metallophore coproporphyrin III in the presence of other microbes.