Summary of Study ST002793
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001741. The data can be accessed directly via it's Project DOI: 10.21228/M8P126 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST002793 |
| Study Title | Interspecies metabolite transfer fuels methionine metabolism of Fusobacterium nucleatum to stimulate volatile methyl mercaptan production |
| Study Summary | The major oral odor compound methyl mercaptan (CH3SH) is strongly associated with halitosis and periodontitis. CH3SH production stems from metabolism of polymicrobial communities in periodontal pockets and on the tongue dorsum. However, understanding of CH3SH-producing oral bacteria and their interactions is limited. This study aimed to investigate CH3SH production by major oral bacteria and the impact of interspecies interactions on its generation. Using a newly constructed large-volume anaerobic non-contact coculture system, Fusobacterium nucleatum was found to be a potent producer of CH3SH, with that production stimulated by metabolic interactions with Streptococcus gordonii, an early dental plaque colonizer. Furthermore, analysis of extracellular amino acids using an S. gordonii arginine-ornithine antiporter (ArcD) mutant demonstrated that ornithine excreted from S. gordonii is a key contributor to increased CH3SH production by F. nucleatum. Further study with 13C, 15N-methionine, as well as gene expression analysis, revealed that ornithine secreted by S. gordonii increased the demand for methionine through accelerated polyamine synthesis by F. nucleatum, leading to elevated methionine pathway activity and CH3SH production. Collectively, these findings suggest that interaction between S. gordonii and F. nucleatum plays a key role in CH3SH production, providing new insight into the mechanism of CH3SH generation in oral microbial communities. Better understanding of the underlying interactions among oral bacteria involved in CH3SH generation can lead to development of more appropriate prophylactic approaches to treat halitosis and periodontitis. |
| Institute | Osaka University |
| Last Name | Kuboniwa |
| First Name | Masae |
| Address | 1-8 Yamadaoka, Suita. Osaka. Japan |
| kuboniwa.masae.dent@osaka-u.ac.jp | |
| Phone | +81668792922 |
| Submit Date | 2023-07-21 |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-07-25 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001741 |
| Project DOI: | doi: 10.21228/M8P126 |
| Project Title: | Interspecies metabolite transfer fuels methionine metabolism of Fusobacterium nucleatum to stimulate volatile methyl mercaptan production |
| Project Summary: | Using 13C/15N-labeled methionine, the fate of methionine in F. nucleatum cells was examined to elucidate the intracellular metabolic dynamics underlying enhanced CH3SH production in the presence of S. gordonii. |
| Institute: | Osaka University |
| Last Name: | Kuboniwa |
| First Name: | Masae |
| Address: | Yamadaoka 1-8, Suita. Osaka, Japan |
| Email: | kuboniwa.masae.dent@osaka-u.ac.jp |
| Phone: | +81668792922 |
Subject:
| Subject ID: | SU002900 |
| Subject Type: | Bacteria |
| Subject Species: | Fusobacterium nucleatum |
| Taxonomy ID: | 190304 |
Factors:
Subject type: Bacteria; Subject species: Fusobacterium nucleatum (Factor headings shown in green)
| mb_sample_id | local_sample_id | Hour |
|---|---|---|
| SA299559 | 0h-2 | 0 |
| SA299560 | 0h-4 | 0 |
| SA299561 | 0h-3 | 0 |
| SA299562 | 1h-4 | 1 |
| SA299563 | 1h-1 | 1 |
| SA299564 | 1h-3 | 1 |
| SA299565 | 2h-4 | 2 |
| SA299566 | 2h-3 | 2 |
| SA299567 | 2h-2 | 2 |
| SA299568 | 3h-1 | 3 |
| SA299569 | 3h-4 | 3 |
| SA299570 | 3h-3 | 3 |
| SA299571 | 6h-4 | 6 |
| SA299572 | 6h-2 | 6 |
| SA299573 | 6h-3 | 6 |
| Showing results 1 to 15 of 15 |
Collection:
| Collection ID: | CO002893 |
| Collection Summary: | To investigate the time course of changes in intracellular metabolites of F. nucleatum cells after coculturing with S. gordonii WT, F. nucleatum cells were collected by centrifugation (7670 × g for 7 min at 4°C), washed twice with 1 ml of ultra-pure water, and then immediately treated twice with 1 ml of 100% methanol containing the internal standards (H3304-1002, Human Metabolome Technologies, Inc., Tsuruoka, Japan). |
| Sample Type: | Bacterial cells |
Treatment:
| Treatment ID: | TR002909 |
| Treatment Summary: | [13C5, 15N] L-Methionine (13C5, 15N, 98%; Taiyo Nippon Sanso Corp., Tokyo, Japan) was added at a final concentration of 10 mM to mCDM, with the following modified amino acid concentrations; 40 mM L-glutamic acid, 10 mM L-arginine-HCl, 1.0 mM L-tryptophan. |
Sample Preparation:
| Sampleprep ID: | SP002906 |
| Sampleprep Summary: | At the mid exponential growth phase (0.5 to 1.0 OD units ml-1), bacterial cells were harvested by centrifugation (7670 × g for 7 min at 4°C), washed twice with PBS, and finally resuspended at 20 OD units ml-1 in mCDM containing 10 mM [13C5, 15N] L-methionine. F. nucleatum and S. gordonii cells were then inoculated at a density of 1.5 × 1010 CFU/well into a Transwell outer chamber (2.6 cm3) and inner chamber (1.5 cm3), respectively. Thereafter anaerobic incubation was performed at 37°C for 0, 1, 2, 3, or 6 h. |
Combined analysis:
| Analysis ID | AN004544 |
|---|---|
| Analysis type | MS |
| Chromatography type | CE |
| Chromatography system | Agilent CE Capillary Electrophoresis System |
| Column | Agilent Fused silica capillary (800 mm x 50 um) |
| MS Type | ESI |
| MS instrument type | Other |
| MS instrument name | Agilent 6210 TOF |
| Ion Mode | POSITIVE |
| Units | pmol/ODml |
Chromatography:
| Chromatography ID: | CH003413 |
| Instrument Name: | Agilent CE Capillary Electrophoresis System |
| Column Name: | Agilent Fused silica capillary (800 mm x 50 um) |
| Column Temperature: | NA |
| Flow Gradient: | NA |
| Flow Rate: | The sample was injected at a pressure of 50 mbar for 10 s (approximately 10 nL) for cation analysis and 25 s (approximately 25 nL) for anion analysis |
| Solvent A: | Solution ID: H3301-1001 for cation analysis |
| Solvent B: | Solution ID: H3302-1021 for anion analysis |
| Chromatography Type: | CE |
MS:
| MS ID: | MS004291 |
| Analysis ID: | AN004544 |
| Instrument Name: | Agilent 6210 TOF |
| Instrument Type: | Other |
| MS Type: | ESI |
| MS Comments: | The systems were controlled by the Agilent G2201AA ChemStation software package, version B.03.01, for CE (Agilent Technologies, Waldbronn, Germany). |
| Ion Mode: | POSITIVE |
