Mass Spectral Molecular Networking of Living Microbial Colonies

Journal Article · Tue Jun 26 00:00:00 EDT 2012 · Proceedings of the National Academy of Sciences of the United States of America, 109(26):E1743-E1752

DOI:https://doi.org/10.1073/pnas.1203689109· OSTI ID:1045109

Watrous, Jeramie D; Roach, Patrick J; Alexandrov, Theodore; Heath, Brandi S; Yang, Jane Y; Kersten, Roland; vander Voort, Menno; Pogliano, Kit; Gross, Harald; Raaijmakers, Jos M; Moore, Bradley S; Laskin, Julia; Bandeira, Nuno; Dorrestein, Pieter C

Integrating the governing chemistry with the genomics and phenotypes of microbial colonies has been a "holy grail" in microbiology. This work describes a highly sensitive, broadly applicable, and costeffective approach that allows metabolic profiling of live microbial colonies directly from a Petri dish without any sample preparation. Nanospray desorption electrospray ionization mass spectrometry (MS), combined with alignment of MS data and molecular networking, enabled monitoring of metabolite production from live microbial colonies from diverse bacterial genera, including Bacillus subtilis, Streptomyces coelicolor, Mycobacterium smegmatis, and Pseudomonas aeruginosa. This work demonstrates that, by using these tools to visualize small molecular changes within bacterial interactions, insights can be gained into bacterial developmental processes as a result of the improved organization of MS/MS data. To validate this experimental platform, metabolic profiling was performed on Pseudomonas sp. SH-C52, which protects sugar beet plants from infections by specific soil-borne fungi [R. Mendes et al. (2011) Science 332:1097–1100]. The antifungal effect of strain SHC52 was attributed to thanamycin, a predicted lipopeptide encoded by a nonribosomal peptide synthetase gene cluster. Our technology, in combination with our recently developed peptidogenomics strategy, enabled the detection and partial characterization of thanamycin and showed that it is amonochlorinated lipopeptide that belongs to the syringomycin family of antifungal agents. In conclusion, the platform presented here provides a significant advancement in our ability to understand the spatiotemporal dynamics of metabolite production in live microbial colonies and communities.

• Research Organization: Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 1045109
• Report Number(s): PNNL-SA-88885; PNASA6; 44679; TRN: US201214%%937
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America, 109(26):E1743-E1752, Vol. 109, Issue 26; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
BACILLUS SUBTILIS
CHEMISTRY
DESORPTION
DETECTION
FUNGI
GENES
IONIZATION
LIGASES
MASS SPECTROSCOPY
METABOLITES
MONITORING
MYCOBACTERIUM
PEPTIDES
PSEUDOMONAS
SAMPLE PREPARATION
STRAINS
STREPTOMYCES
SUGAR BEETS
Environmental Molecular Sciences Laboratory