Fermentation Associated Changes in Phenolic Profile, Antioxidant Capacity, and In Vitro Apoptotic Activity of Soursop Leaf Kombucha Against T47D Breast Cancer Cells
DOI:
https://doi.org/10.48048/tis.2026.12886Keywords:
Soursop leaf kombucha, Fermentation, Antioxidant activity, Anticancer potentialAbstract
Kombucha fermentation of soursop leaves is associated with changes in functional properties through post-fermentation physicochemical and metabolite profile modifications. This study was evaluated for its effects on physicochemical properties, qualitative LC-MS metabolite profiles, antioxidant capacity, and in vitro anticancer activity against T47D breast cancer cells. Fermentation was carried out for 12 days using leaf concentrations of 0.6% - 2.0%, followed by measurements of pH, total microbial counts, total phenolic and flavonoid contents, antioxidant activity, and LC-MS metabolite profiling. Cytotoxicity and apoptosis induction were assessed using CCK-8 assays, morphological observations, fluorescence imaging (DAPI, Annexin V-FITC/PI), and flow cytometry. At a 1.4% leaf concentration, endpoint comparison between day 0 and day 12 showed an increase in total viable counts from 6.77 to 7.65 log CFU/mL and a marked increase in total phenolic content from 198.13 to 322.03 mg GAE/mL, accompanied by the greatest reduction in antioxidant IC₅₀ values (from 252.29 to 99.73 ppm). Qualitative LC-MS profiling revealed differences in detected compounds between fermented and unfermented samples, including the presence of aglycone flavonoids in fermented kombucha. Acidic soursop kombucha demonstrated the strongest cytotoxicity toward T47D cells (IC₅₀ = 2.35 mg/mL), followed by neutralized kombucha (IC₅₀ = 3.87 mg/mL) and soursop leaf drink (IC₅₀ = 7.37 mg/mL). Flow cytometry indicated a marked shift toward apoptotic populations in fermented samples, with combined early and late apoptosis reaching 83.56% in acidic kombucha treated cells. Kombucha fermentation under acidic conditions was associated with enhanced antioxidant capacity and in vitro anticancer related activity of soursop leaves.
HIGHLIGHTS
- Kombucha fermentation significantly changed physicochemical properties and metabolite profiles.
- Optimal leaf concentrations (1.4%) were identified using Zeleny’s method based on pre–post fermentation improvements in total phenolics and antioxidant IC₅₀.
- Qualitative LC-MS showed broader metabolite profiles after fermentation with improved bioactivity.
- Soursop leaf kombucha showed the strongest cytotoxicity toward T47D breast cancer cells (IC₅₀ = 2.35 mg/ml).
- Fermented samples induced higher early and late apoptosis, supporting their chemopreventive potential.
GRAPHICAL ABSTRACT
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