Postbiotic cell free supernatant from Lactobacillus strain: Isolation, characterization, In vitro assessment of antimicrobial and cytotoxic potentials using Vero cell line

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Research Paper 15/07/2025
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Postbiotic cell free supernatant from Lactobacillus strain: Isolation, characterization, In vitro assessment of antimicrobial and cytotoxic potentials using Vero cell line

Arunavarsini Kumarasamy, R. Sornambiga, Mahenthiran Ramasamy
Int. J. Biosci. 27(1), 297-305, July 2025.
Copyright Statement: Copyright 2025; The Author(s).
License: CC BY-NC 4.0

Abstract

This study investigates the isolation, identification, and characterization of a Lactobacillus paracasei strain derived from homemade yogurt, emphasizing its potential as a source of bioactive postbiotics. Using morphological and biochemical methods, including MALDI-TOF MS, the strain was reliably identified at the genus and species levels. The probiotic nature of the isolate was confirmed through culture and microscopic analysis. The research further explored the production of postbiotic cell-free supernatants from the isolated strain, assessing their antimicrobial and cytotoxic activities. The postbiotics demonstrated significant inhibitory effects against pathogenic bacteria such as Escherichia coli, Salmonella enterica, Streptococcus bovis, and Enterococcus faecalis, often exhibiting antimicrobial efficacy comparable or superior to standard antibiotics. The safety profile was affirmed by cytotoxicity assays on Vero cell lines, underscoring their low toxicity and potential for safe therapeutic application. The findings support the notion that postbiotics hold promising prospects as natural antimicrobial agents, especially in light of rising antibiotic resistance. Their stability and ease of handling offer advantages over live probiotics. The study emphasizes the importance of further clinical investigations and mechanistic studies to optimize the application of postbiotics in medical and food industry contexts, promoting safer and more effective health interventions.

Aghebati-Maleki L, Hasannezhad P, Abbasi A, Khani N. 2021. Antibacterial, antiviral, antioxidant, and anticancer activities of postbiotics: A review of mechanisms and therapeutic perspectives. Biointerface Research in Applied Chemistry 12(2), 2629–2645. https://doi.org/10.33263/BRIAC122.26292645.

Ammerman NC, Beier-Sexton M, Azad AF. 2008. Growth and maintenance of Vero cell lines. Current Protocols in Microbiology 11(1), A-4E. https://doi.org/10.1002/9780471729259.mca04es11.

Ariful H, Saiful H, Dipa R, Zohorul IM. 2023. Screening, isolation and evaluation of probiotic potential Lactobacillus acidophilus strains from available sources in Bangladesh. International Journal of Biosciences 23(1), 68–80. http://dx.doi.org/10.12692/ijb/23.1.68-80.

Bourebaba Y, Marycz K, Mularczyk M, Bourebaba L. 2022. Postbiotics as potential new therapeutic agents for metabolic disorders management. Biomedicine & Pharmacotherapy 153, 113138. https://doi.org/10.1016/j.biopha.2022.113138.

Dahlan HA, Sani NA. 2017. The interaction effect of mixing starter cultures on homemade natural yogurt’s pH and viscosity. International Journal of Food Studies 6(2), 152–158. https://doi.org/10.7455/ijfs/6.2.2017.a3.

Elham N, Naheed M, Elahe M, Hossein MM, Majid T. 2022. Selective cytotoxic effect of probiotic, paraprobiotic and postbiotics of L. casei strains against colorectal cancer cells: In vitro studies. Brazilian Journal of Pharmaceutical Sciences 58, e19400. http://dx.doi.org/10.1590/s2175-97902022e19400.

Hijová E. 2024. Postbiotics as metabolites and their biotherapeutic potential. International Journal of Molecular Sciences 25(10), 5441. https://doi.org/10.3390/ijms25105441.

Kaewchomphunuch T, Charoenpichitnunt T, Thongbaiyai V, Ngamwongsatit N, Kaeoket K. 2022. Cell-free culture supernatants of Lactobacillus spp. and Pediococcus spp. inhibit growth of pathogenic Escherichia coli isolated from pigs in Thailand. BMC Veterinary Research 18(1), 60. https://doi.org/10.1186/s12917-022-03140-8.

Karami S, Roayaei M, Hamzavi H, Bahmani M, Hassanzad-Azar H, Leila M, Rafieian-Kopaei M. 2017. Isolation and identification of probiotic Lactobacillus from local dairy and evaluating their antagonistic effect on pathogens. International Journal of Pharmaceutical Investigation 7(3), 137–141. https://dx.doi.org/10.4103/jphi.JPHI_8_17.

Lee JY, Kim Y, Kim JI, Lee HY, Moon GS, Kang CH. 2022. Improvements in human keratinocytes and antimicrobial effect mediated by cell-free supernatants derived from probiotics. Fermentation 8(7), 332. https://doi.org/10.3390/fermentation8070332.

Mani LE, Daniela AB, Aurelio LM. 2022. The impacts of antimicrobial and antifungal activity of cell‐free supernatants from lactic acid bacteria in vitro and foods. Comprehensive Reviews in Food Science and Food Safety 21(1), 604–641. https://doi.org/10.1111/1541-4337.12872.

Olson DW, Aryana KJ. 2022. Probiotic incorporation into yogurt and various novel yogurt-based products. Applied Sciences 12, 12607. https://doi.org/10.3390/app122412607.

Ozbey G, Cambay Z, Yilmaz S, Aytekin O, Zigo F, Ozçelik M, Otlu B. 2022. Identification of bacterial species in milk by MALDI-TOF and assessment of some oxidant-antioxidant parameters in blood and milk from cows with different health status of the udder. Polish Journal of Veterinary Sciences 25(2), 269–277. https://doi.org/10.24425/pjvs.2022.141811.

Phani Kumari YV, Kavita W. 2024. Isolation and identification of probiotic bacteria from natural Neera to extend the shelf life of fresh fruits and vegetables. International Journal of Biosciences 24(5), 25–37. http://dx.doi.org/10.12692/ijb/24.5.25-37.

Piaru SP, Mahmud R, Perumal S. 2012. Determination of antibacterial activity of essential oil of Myristica fragrans Houtt. using tetrazolium microplate assay and its cytotoxic activity against Vero cell line. International Journal of Pharmacology 8(6), 572–576. https://doi.org/10.3923/ijp.2012.572.576.

Thorakkattu P, Khanashyam AC, Shah K, Babu KS, Mundanat AS, Deliephan A, Deokar GS, Santivarangkna C, Nirmal NP. 2022. Postbiotics: Current trends in food and pharmaceutical industry. Foods 11(19), 3094. https://doi.org/10.3390/foods11193094.

Widiastuti NK, Virginia NM, Yastawan I, Permatasari AAAP, Wiradana PA, Widhiantara IG, Sucipto TH. 2023. Cytotoxicity evaluation of Erythrina lithosperma Miq. leaf extract against Vero cell lines: In vitro study. Research Journal of Pharmacy and Technology 16(1), 153–158. http://dx.doi.org/10.52711/0974-360X.2023.00028.

Yang KM, Kim JS, Kim HS, Kim YY, Oh JK, Jung HW, Park DS, Bae KH. 2021. Lactobacillus reuteri AN417 cell-free culture supernatant as a novel antibacterial agent targeting oral pathogenic bacteria. Scientific Reports 11(1), 1631. https://doi.org/10.1038/s41598-020-80921-x.

Zendeboodi F, Khorshidian N, Mortazavian AM, DA Cruz AG. 2020. Probiotic: Conceptualization from a new approach. Current Opinion in Food Science 32, 103–123. https://doi.org/10.1016/j.cofs.2020.03.009.

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