Summary of Study ST001433
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 PR000984. The data can be accessed directly via it's Project DOI: 10.21228/M8H68N This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
Study ID | ST001433 |
Study Title | Evidence for proline utilisation by oral bacterial biofilms grown in saliva |
Study Type | Research study |
Study Summary | Within the mouth bacteria are starved of saccharides as their main nutrient source between meals and it is unclear what drives their metabolism. Previously oral in vitro biofilms grown in saliva have shown proteolytic degradation of salivary proteins and increased extracellular proline. Although arginine and glucose have been shown before to have an effect on oral biofilm growth and activity, there is limited evidence for proline. Nuclear magnetic resonance (NMR) spectroscopy was used to identify extracellular metabolites produced by bacteria in oral biofilms grown on hydroxyapatite discs. Biofilms were inoculated with whole mouth saliva and then grown for 7 days using sterilised whole mouth saliva supplemented with proline, arginine and glucose as a growth-medium. Overall proline had a beneficial effect on biofilm growth – with significantly fewer dead bacteria present by biomass and surface area of the biofilms (p <0.05). Where arginine and glucose significantly increased and decreased pH, respectively, the pH of proline supplemented biofilms remained neutral at pH 7.3-7.5. SDS-polyacrylamide gel electrophoresis of the spent saliva from proline and arginine supplemented biofilms showed inhibition of salivary protein degradation of immature biofilms. NMR analysis of the spent saliva revealed that proline supplemented biofilms were metabolically similar to unsupplemented biofilms, but these biofilms actively metabolised proline to 5-aminopentanoate, butyrate and propionate, and actively utilised glycine. This study shows that in a nutrient limited environment, proline has a beneficial effect on in vitro oral biofilms grown from a saliva inoculum. |
Institute | King's College London |
Department | Centre for Host Microbiome Interactions |
Last Name | Cleaver |
First Name | Leanne |
Address | Floor 17, Tower Wing, Guy's Hospital, Great Maze Pond |
leanne.cleaver@kcl.ac.uk | |
Phone | 07464626438 |
Submit Date | 2020-07-23 |
Num Groups | 11 |
Raw Data Available | Yes |
Raw Data File Type(s) | raw(NMR) |
Analysis Type Detail | NMR |
Release Date | 2020-08-03 |
Release Version | 1 |
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Sample Preparation:
Sampleprep ID: | SP001515 |
Sampleprep Summary: | Samples were processed for NMR using a previously published method (6). Briefly, centrifuged spent saliva supernatant and the sterile saliva sample were each mixed with TSP buffer in a 5 mm NMR tube (Bruker, Germany). The tubes were sealed and analysed at the Biomolecular Spectroscopy Centre, King’s College London, UK on a 600 MHz spectrometer (Bruker) for 1H 1D-NMR and 1H-C13 1D- and 2D-NMR. The concentration of metabolites relative to the sterile saliva baseline were collected using Chenomix NMR Suite version 8.5 (Chenomix Ltd, Canada). The 2D C13 spectra were analysed using TopSpin version 3.6.2 (Bruker) and COLMAR (30). |