Apr 28, 2016

Localizing metabolic synthesis in microbial cultures with kinetic mass spectrometry imaging (kMSI)

BioRxiv : the Preprint Server for Biology
Katherine B LouieTrent R Northen

Abstract

Mass spectrometry imaging (MSI) has emerged as a powerful technique enabling spatially defined imaging of metabolites within microbial biofilms. Here, we extend this approach to enable differentiation of newly synthesized versus pre-existing metabolites across a co-culture. This is accomplished by MS imaging two soil microbes, Shewanella oneidensis MR1 and Pseudomonas stutzeri RCH2, that were administered heavy water (D2O) during growth on agar plates. For two species-specific diglyceride (DG) lipids, isotopic analysis was performed on each spectra collected across the co-culture to determine the relative amount of newly synthesized versus pre-existing lipid. Here, highest levels of new synthesis of RCH2 lipid was localized to border regions adjacent to S. oneidensis MR1, while the MR1 lipid showed highest levels in regions further from RCH2. Interestingly, regions of high lipid abundance did not correspond to the regions with highest new lipid biosynthesis. Given the simplicity and generality of using D2O as a stable isotopic probe combined with the accessibility of kMSI to a range of MSI instrumentation, this approach has broad application for improving our understanding of how microbial interactions influence metabolite bios...Continue Reading

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Mentioned in this Paper

Metabolic Process, Cellular
Lipogenesis
Agar
Coculture Techniques
Microbial Interactions
Cell Differentiation Process
Pseudomonas stutzeri
Mass Spectrometry
Defined Imaging
Microbial

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