{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T13:42:10Z","timestamp":1787060530585,"version":"build-2736575974"},"reference-count":37,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2025,8,29]],"date-time":"2025-08-29T00:00:00Z","timestamp":1756425600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Center for Absorption in the Science Ministry of Immigration and Absorption, Israel"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Streptococcus mutans (S. mutans) is a cariogenic bacterium in the oral cavity that plays a significant role in plaque formation and dental caries. In previous research by our group, we showed that the endocannabinoid anandamide (AEA) has anti-bacterial and anti-biofilm activities against S. mutans. Here, we aimed to investigate its effects on S. mutans through metabolomics analyses. S. mutans was cultivated in the absence or presence of AEA at a sub-minimum inhibitory concentration (MIC), and changes in metabolites and metabolic pathways were assessed through liquid chromatography\u2013mass spectrometry (LC-MS). Treatment of S. mutans using AEA at 10 \u00b5g\/mL significantly disturbed the glycolytic flux in the bacteria, which was indicated by a reduced glucose uptake into the cell, suppression of key glycolytic intermediates, reduced acid production into the media, imbalance of NAD+\/NADH, and decreased adenosine triphosphate (ATP) production. The disruption of carbohydrate metabolism impacts critical cellular processes, including energy production, redox balance, and biosynthetic pathways, leading to metabolic stress and impaired cellular function. These results highlight the mode of action of AEA as an antimicrobial agent. Altogether, these findings suggest that AEA has potential as a novel antimicrobial agent in the development of therapeutics against S. mutans.<\/jats:p>","DOI":"10.3390\/ijms26178401","type":"journal-article","created":{"date-parts":[[2025,8,29]],"date-time":"2025-08-29T16:42:21Z","timestamp":1756485741000},"page":"8401","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Anandamide Alters Glycolytic Activity in Streptococcus mutans: Metabolomics and Stable Isotope Labeling Study"],"prefix":"10.3390","volume":"26","author":[{"given":"Goldie","family":"Wolfson","sequence":"first","affiliation":[{"name":"Institute of Biomedical and Oral Research (IBOR), Faculty of Dental Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"},{"name":"Department of Clinical Microbiology and Infectious Diseases, Hadassah-Hebrew University Medical Center, Jerusalem 9112001, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Doron","family":"Steinberg","sequence":"additional","affiliation":[{"name":"Institute of Biomedical and Oral Research (IBOR), Faculty of Dental Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandra","family":"Eliassaf","sequence":"additional","affiliation":[{"name":"The Metabolic Profiling Unit, The Core Research Facility, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Morshina","sequence":"additional","affiliation":[{"name":"The Metabolic Profiling Unit, The Core Research Facility, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5944-6555","authenticated-orcid":false,"given":"C\u00e9sar Jess\u00e9","family":"Enr\u00edquez-Rodr\u00edguez","sequence":"additional","affiliation":[{"name":"The Metabolic Profiling Unit, The Core Research Facility, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"},{"name":"Respiratory Medicine Department, Hospital del Mar Research Institute-Hospital del Mar-BRN, 08003 Barcelona, Spain"},{"name":"Medicine and Life Sciences Department, Universitat Pompeu Fabra (UPF), 08003 Barcelona, Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red, \u00c1rea de Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, 28029 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4325-5260","authenticated-orcid":false,"given":"Itzhack","family":"Polacheck","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology and Infectious Diseases, Hadassah-Hebrew University Medical Center, Jerusalem 9112001, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3276-0963","authenticated-orcid":false,"given":"Maya","family":"Korem","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology and Infectious Diseases, Hadassah-Hebrew University Medical Center, Jerusalem 9112001, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ori","family":"Shalev","sequence":"additional","affiliation":[{"name":"The Metabolic Profiling Unit, The Core Research Facility, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112102, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Sionov, R.V., and Steinberg, D. (2022). Anti-Microbial Activity of Phytocannabinoids and Endocannabinoids in the Light of Their Physiological and Pathophysiological Roles. Biomedicines, 10.","DOI":"10.3390\/biomedicines10030631"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1946","DOI":"10.1126\/science.1470919","article-title":"Isolation and Structure of a Brain Constituent That Binds to the Cannabinoid Receptor","volume":"258","author":"Devane","year":"1992","journal-title":"Science"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1016\/j.tins.2017.04.005","article-title":"Endocannabinoid Signaling in the Control of Social Behavior","volume":"40","author":"Wei","year":"2017","journal-title":"Trends Neurosci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3178","DOI":"10.1038\/s41380-020-00905-1","article-title":"Role of Endocannabinoid Signaling in a Septohabenular Pathway in the Regulation of Anxiety- and Depressive-like Behavior","volume":"26","author":"Vickstrom","year":"2021","journal-title":"Mol. Psychiatry"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1038\/s41401-018-0051-5","article-title":"Integrating Endocannabinoid Signaling in the Regulation of Anxiety and Depression","volume":"40","author":"Yin","year":"2019","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"8690","DOI":"10.1038\/s41598-021-88099-6","article-title":"Anandamide Alters the Membrane Properties, Halts the Cell Division and Prevents Drug Efflux in Multidrug Resistant Staphylococcus aureus","volume":"11","author":"Banerjee","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"17696","DOI":"10.1038\/s41598-018-35793-7","article-title":"Antimicrobial Potential of Endocannabinoid and Endocannabinoid-like Compounds against Methicillin-Resistant Staphylococcus aureus","volume":"8","author":"Feldman","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Sionov, R.V., Banerjee, S., Bogomolov, S., Smoum, R., Mechoulam, R., and Steinberg, D. (2022). Targeting the Achilles\u2019 Heel of Multidrug-Resistant Staphylococcus aureus by the Endocannabinoid Anandamide. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23147798"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"9371","DOI":"10.1073\/pnas.161191698","article-title":"Supersensitivity to Anandamide and Enhanced Endogenous Cannabinoid Signaling in Mice Lacking Fatty Acid Amide Hydrolase","volume":"98","author":"Cravatt","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1038\/s41564-019-0655-7","article-title":"Growth Effects of N-Acylethanolamines on Gut Bacteria Reflect Altered Bacterial Abundances in Inflammatory Bowel Disease","volume":"5","author":"Fornelos","year":"2020","journal-title":"Nat. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"13728","DOI":"10.1038\/s41598-020-70650-6","article-title":"Anandamide Prevents the Adhesion of Filamentous Candida albicans to Cervical Epithelial Cells","volume":"10","author":"Sionov","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Wolfson, G., Sionov, R.V., Smoum, R., Korem, M., Polacheck, I., and Steinberg, D. (2023). Anti-Bacterial and Anti-Biofilm Activities of Anandamide against the Cariogenic Streptococcus mutans. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24076177"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1111\/omi.12162","article-title":"Acid-Adaptive Mechanisms of Streptococcus mutans\u2013the More We Know, the More We Don\u2019t","volume":"32","author":"Baker","year":"2017","journal-title":"Mol. Oral Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1099\/13500872-142-1-33","article-title":"Lactic Acid Excretion by Streptococcus mutans","volume":"142","author":"Dashper","year":"1996","journal-title":"Microbiology"},{"key":"ref_15","first-page":"10","article-title":"The Biology of Streptococcus Mutans","volume":"7","author":"Lemos","year":"2018","journal-title":"Microbiol. Spectr."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1007\/s10096-013-1993-7","article-title":"The Virulence of Streptococcus mutans and the Ability to Form Biofilms","volume":"33","author":"Jurczak","year":"2014","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3682","DOI":"10.1021\/acs.jproteome.4c00382","article-title":"Trans-Cinnamaldehyde Inhibition of Pyruvate Dehydrogenase: Effects on Streptococcus mutans Carbohydrate Metabolism","volume":"23","author":"Zhang","year":"2024","journal-title":"J. Proteome Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1034\/j.1399-302X.2003.00068.x","article-title":"Xylitol Inhibition of Anaerobic Acid Production by Streptococcus mutans at Various PH Levels","volume":"18","author":"Miyasawa","year":"2003","journal-title":"Oral Microbiol. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2056291","DOI":"10.1080\/20002297.2022.2056291","article-title":"Nicotinamide Could Reduce Growth and Cariogenic Virulence of Streptococcus mutans","volume":"14","author":"Lin","year":"2022","journal-title":"J. Oral Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1159\/000088190","article-title":"Synergistic Inhibition by Combination of Fluoride and Xylitol on Glycolysis by Mutans Streptococci and Its Biochemical Mechanism","volume":"39","author":"Maehara","year":"2005","journal-title":"Caries Res."},{"key":"ref_21","first-page":"76","article-title":"A Review On Streptococcus mutans With Its Diseases Dental Caries, Dental Plaque And Endocarditis","volume":"2","author":"Daboor","year":"2015","journal-title":"Indian J. Microbiol. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2816812","DOI":"10.1155\/2016\/2816812","article-title":"Influence of the Culture Medium in Dose-Response Effect of the Chlorhexidine on Streptococcus mutans Biofilms","volume":"2016","author":"Queiroz","year":"2016","journal-title":"Scientifica"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Chamlagain, M., Hu, J., Sionov, R.V., and Steinberg, D. (2024). Anti-Bacterial and Anti-Biofilm Activities of Arachidonic Acid against the Cariogenic Bacterium Streptococcus Mutans. Front. Microbiol., 15.","DOI":"10.3389\/fmicb.2024.1333274"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Rewak-Soroczynska, J., Dorotkiewicz-Jach, A., Drulis-Kawa, Z., and Wiglusz, R.J. (2022). Culture Media Composition Influences the Antibacterial Effect of Silver, Cupric, and Zinc Ions against Pseudomonas aeruginosa. Biomolecules, 12.","DOI":"10.3390\/biom12070963"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"368","DOI":"10.1111\/lam.12863","article-title":"Growth Media and Assay Plate Material Can Impact on the Effectiveness of Cationic Biocides and Antibiotics against Different Bacterial Species","volume":"66","author":"Bock","year":"2018","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1177\/00220345920710050601","article-title":"pH Regulation by Streptococcus mutans","volume":"71","author":"Dashper","year":"1992","journal-title":"J. Dent. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2751","DOI":"10.1128\/jb.177.10.2751-2759.1995","article-title":"Regulation of ATP-Dependent P-(Ser)-HPr Formation in Streptococcus mutans and Streptococcus salivarius","volume":"177","author":"Thevenot","year":"1995","journal-title":"J. Bacteriol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"647611","DOI":"10.3389\/fmicb.2021.647611","article-title":"In Vitro Antibacterial Activity of an FDA-Approved H+-ATPase Inhibitor, Bedaquiline, Against Streptococcus mutans in Acidic Milieus","volume":"12","author":"Zhang","year":"2021","journal-title":"Front. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.abb.2019.03.014","article-title":"Proton-Pumping F-ATPase Plays an Important Role in Streptococcus mutans under Acidic Conditions","volume":"666","author":"Sekiya","year":"2019","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3645","DOI":"10.1128\/JB.00383-15","article-title":"Loss of NADH Oxidase Activity in Streptococcus mutans Leads to Rex-Mediated Overcompensation in NAD+ Regeneration by Lactate Dehydrogenase","volume":"197","author":"Baker","year":"2015","journal-title":"J. Bacteriol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3748","DOI":"10.1128\/JB.00169-06","article-title":"Different Roles of EIIABMan and EIIGlc in Regulation of Energy Metabolism, Biofilm Development, and Competence in Streptococcus mutans","volume":"188","author":"Abranches","year":"2006","journal-title":"J. Bacteriol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1111\/j.1399-302X.1995.tb00167.x","article-title":"Mechanism of Inhibition of Glycolysis in Streptococcus mutans NCIB 11723 by Chlorhexidine","volume":"10","author":"Iwami","year":"1995","journal-title":"Oral Microbiol. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1139\/w06-055","article-title":"Triclosan Inhibition of Membrane Enzymes and Glycolysis of Streptococcus mutans in Suspensions and Biofilms","volume":"52","author":"Phan","year":"2006","journal-title":"Can. J. Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"20859","DOI":"10.1038\/s41598-021-99873-x","article-title":"Comparison of Bacteria Disintegration Methods and Their Influence on Data Analysis in Metabolomics","volume":"11","author":"Mielko","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/bs.mie.2015.05.016","article-title":"Analysis of Cell Metabolism Using LC-MS and Isotope Tracers","volume":"Volume 561","author":"MacKay","year":"2015","journal-title":"Methods Enzymol"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3330","DOI":"10.1128\/AEM.66.8.3330-3336.2000","article-title":"Genotypic Heterogeneity of Streptococcus oralis and Distinct Aciduric Subpopulations in Human Dental Plaque","volume":"66","author":"Alam","year":"2000","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2382","DOI":"10.1128\/AEM.68.5.2382-2390.2002","article-title":"Analysis of Streptococcus mutans Proteins Modulated by Culture under Acidic Conditions","volume":"68","author":"Wilkins","year":"2002","journal-title":"Appl. Environ. Microbiol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/17\/8401\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:35:11Z","timestamp":1760034911000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/17\/8401"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,29]]},"references-count":37,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["ijms26178401"],"URL":"https:\/\/doi.org\/10.3390\/ijms26178401","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,29]]}}}