Human vaginal Lactobacillus Jensenii -derived (-)-Terpinen-4-ol restores antibiotic sensitivity by inhibiting efflux pumps in drug resistant E. coli and K. pneumoniae
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Abstract
Probiotics present a promising preventive strategy for addressing urinary tract infections (UTIs) and serve as a viable alternative to conventional antimicrobial treatments. The integration of Lactobacillus-derived efflux pump inhibitors (EPIs) with conventional antibiotics presents a promising strategy to enhance the efficacy of treatments against antibiotic-resistant superbugs. In this study, we isolated eight vaginal Lactobacillus spp. from 54 healthy Indian women to explore their probiotic properties, specifically the ability of cell-free supernatant (CFS) from Lactobacillus jensenii. to inhibit efflux in Multi Drug Resistant (MDR) E. coli and K. pneumoniae. From the CFS, we obtained seven fractions, among which compound 7 (C7) markedly reduced the MIC of erythromycin against the K. pneumoniae MTCC 432 strain by 32-fold from 64 to 2 µg and restored its sensitivity, indicating potent efflux inhibitory activity. Gas Chromatography-Mass Spectrometry (GC-MS) analysis identified one major compound from the C7 fraction as Cyclohexen-1-ol, 4-methyl-1-(1-methylethyl)-, (R)-, also known as (-)-Terpinen-4-ol. In vivo studies using zebrafish infected with clinical strain of K. pneumoniae 1845 followed by treatment with (-)-Terpinen-4-ol and erythromycin caused a significant 7-log reduction in bacterial bioburden. Additionally, time-kill assay demonstrated that the combination of erythromycin with (-)-Terpinen-4-ol caused a remarkable 7-log reduction in both K. pneumoniae, and E. coli cell counts. Furthermore, (-)-Terpinen-4-ol -erythromycin combination exhibited a six-log reduction in bacterial burden in infected T24 bladder cell lines and was found to be non-toxic. This study underscores the efflux inhibitory potential of (-)-Terpinen-4-ol extracted from the CFS of human vaginal probiotics, paving the way for future pharmacological research and therapeutic applications.
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Lactobacilli-derived terpenes have been demonstrated to inhibit efflux pumps in drug-resistant pathogens.
“Lactobacilli-derived terpenes have been recently demonstrated to inhibit efflux pumps in drug-resistant pathogens [83].”
From Bile Salt Hydrolase Activity in the Food-Derived Strain Levilactobacillus brevis M3R3: Genomic and Functional Characterization (2026) · cites this paper as [83]
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Agolino, G., Cristofolini, M., Ronsivalle, M. A., Cattivelli, A., Tagliazucchi, D., Caggia, C., Solieri, L., & Randazzo, C. L. (2026) · Probiotics and Antimicrobial Proteins · doi:10.1007/s12602-025-10810-0 · Cited by 1 · see citing works
“Lactobacilli-derived terpenes have been recently demonstrated to inhibit efflux pumps in drug-resistant pathogens [83].”
- Authors
- Nair, V. G., Unnikrishnan, V. K., Muralidharan, N., Venkatathri, B. N. S., Lowrence, R. C., YBRD, R., Narbhavi, D., A, A., N, P., & Nagarajan, S.
- Affiliations
- SASTRA University (IN)
- Corresponding author
- Nagarajan, Saisubramanian
- Venue
- Scientific Reports
- Published
- 2025-08-29
- Volume
- 15
- Issue
- 1
- Pages
- 31823-31823
- ISSN
- 2045-2322 (ISSN-L)
- PMC
- PMC12397448
- OpenAlex
- W4413802848
- Cited by
- 8
- References
- 78
- Citations by year
- 2026: 7 · 2025: 1
- FWCI
- 2.25
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Scientific Reports (publishedVersion, cc-by-nc-nd)
- Research Square (acceptedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Department of Science and Technology, Ministry of Science and Technology, India (DST/WOS-B/HN-32/2021)
- Indian Council of Medical Research (OMI/03/2022/ECD, EMDR/SG/14/2024/01-04170)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Antibiotic Resistance in Bacteria
- Life Sciences › Agricultural and Biological Sciences › Food Science › Probiotics and Fermented Foods
- Life Sciences › Agricultural and Biological Sciences › Food Science › Essential Oils and Antimicrobial Activity
- Keywords
- Lactobacillus jensenii, terpinen-4-ol, efflux pump inhibitors, antibiotic resistance, multidrug-resistant Escherichia coli, multidrug-resistant Klebsiella pneumoniae, probiotics, cell-free supernatant, erythromycin, urinary tract infection, combination therapy, vaginal microbiota, zebrafish infection model, bladder cancer cell lines
- MeSH terms
- Animals, Anti-Bacterial Agents, Erythromycin, Escherichia coli, Female, Humans, Klebsiella pneumoniae, Lactobacillus, Microbial Sensitivity Tests, Vagina, Zebrafish, Probiotics, Drug Resistance, Multiple, Bacterial
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