Methylotroph Quorum Sensing Signal Identification by Inverse Stable Isotopic Labeling
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Abstract
Abstract Natural products are an essential source of bioactive compounds. Isotopic labeling is an effective way to identify natural products that incorporate a specific precursor; however, this approach is limited by the availability of isotopically-enriched precursors. We used an inverse stable isotopic labeling approach to identify natural products by growing bacteria on a 13 C-carbon source and then identifying 12 C-precursor incorporation by mass spectrometry. We applied this approach to methylotrophs, ecologically important bacteria predicted to have significant yet underexplored biosynthetic potential. We demonstrate this method identifies N -acyl homoserine lactone quorum sensing signals produced by diverse methylotrophs grown on three different one-carbon compounds. We then apply this approach to simultaneously identify five uncharacterized signals produced by a methylotroph, and link these compounds to their synthases. We envision that this method can be used to identify other natural product classes synthesized by methylotrophs and other organisms that grow on relatively inexpensive 13 C-carbon sources.
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Cummings et al. used INVERSIL for the discovery and structure elucidation of a set of new homoserine lactones.
“Cummings et al. used this technique for the discovery and structure elucidation of a set of new homo serine lactones.”
From Connecting metabolome and phenotype: recent advances in functional metabolomics tools for the identification of bioactive natural products (2024) · cites this paper as [71]
In “Simultaneous identification of five acyl-HSL signals produced by the methylotroph DSM2163” (page unavailable), the authors report using inverse stable isotopic labeling to identify five previously unidentified signals and confirming their identities by mass spectrometry and comparison with standards or previously characterized signals. The signals were newly identified in DSM2163, but the passage does not present them as new homoserine lactone compounds whose structures were newly elucidated. (LLM, conf=0.88)
Experimental — accuracy not yet measuredINVERSIL is an inverse stable isotope labeling technique typically used for organisms that grow on minimal media with limited carbon and nitrogen source.
“a key advance in stable isotope studies is the inverse stable isotope labeling technique, termed INVERSIL.71 With this method, typically used for organism that grow on minimal media with limited carbon and nitrogen source, the organism is cultured with 13C and/or 15N as sole carbon and nitrogen source.”
From Connecting metabolome and phenotype: recent advances in functional metabolomics tools for the identification of bioactive natural products (2024) · cites this paper as [71]
The Abstract (page unavailable) describes inverse stable isotopic labeling using a 13C-grown organism and an unlabeled precursor. The discussion under “Heterologous expression links acyl-HSL signals with their synthases” (page unavailable) says the method is especially well-suited to organisms that grow on inexpensive 13C-carbon sources, but does not say it is typically used with minimal media or limited carbon and nitrogen sources. Our copy also does not name the method “INVERSIL” or describe 15N labeling as part of it. (LLM, conf=0.88)
Experimental — accuracy not yet measured
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- Authors
- Cummings, D. A., Snelling, A. I., & Puri, A. W.
- Affiliations
- University of Utah (US)
- Corresponding author
- Puri, Aaron W.
- Venue
- ACS Chemical Biology
- Publisher
- American Chemical Society (ACS)
- Published
- 2021-07-30
- Volume
- 16
- Issue
- 8
- Pages
- 1332-1338
- ISSN
- 1554-8929 (ISSN-L)
- 1554-8937
- PMC
- PMC8496459
- OpenAlex
- W3184516418
- Cited by
- 12
- References
- 43
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 5 · 2023: 3
- FWCI
- 1.17
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Licence
- Locations
- ACS Chemical Biology (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of General Medical Sciences (R00 GM118762)
- Topics
- Health Sciences › Medicine › Pharmacology › Microbial Natural Products and Biosynthesis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bacterial biofilms and quorum sensing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial metabolism and enzyme function
- Keywords
- methylotrophic bacteria, quorum sensing, N-acyl homoserine lactones, precursor incorporation, natural product identification, biosynthetic gene identification, one-carbon compounds, bacterial secondary metabolites
- MeSH terms
- Proof of Concept Study, Carbon, Carbon Isotopes, Isotope Labeling, Methylococcaceae, Methylobacteriaceae, Quorum Sensing, Acyl-Butyrolactones
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