January 13, 2023
Journal Article

Balancing trade-offs imposed by growth media and mass spectrometry for bacterial exometabolomics

Abstract

The bacterial exometabolome consists of a vast array of specialized metabolites, many of which are only produced in response to specific environmental stimuli. For this reason, it is desirable to control the extracellular environment with a defined growth medium composed of pure ingredients. However, complex (undefined) media are expected to support the robust growth of a greater variety of microorganisms than defined media. Here, we investigate the trade-offs inherent to a range of complex and defined solid media for the growth of soil microorganisms, production of specialized metabolites, and detection of these compounds using direct infusion mass spectrometry. We find that complex media support growth of more soil microorganisms, as well as allowing for the detection of more previously discovered natural products as a fraction of total m/z features detected in each sample. However, the use of complex media often caused mass spectrometer injection failures and poor-quality mass spectra, which in some cases resulted in over a quarter of samples being removed from analysis. Defined media, while more limiting in growth, generated higher quality spectra and yielded more m/z features after background subtraction. These results inform future exometabolomic experiments requiring a medium that supports the robust growth of many soil microorganisms.

Published: January 13, 2023

Citation

Donnelly A.E., N. Narayanan, C. Birer, T.J. DeWolfe, R.K. Chu, C.R. Anderton, and E.S. Wright. 2022. Balancing trade-offs imposed by growth media and mass spectrometry for bacterial exometabolomics. Applied and Environmental Microbiology 88, no. 20. PNNL-SA-169988. doi:10.1128/aem.00922-22

Research topics