Objective: This study aimed to investigate the antioxidant properties of kombucha prepared from Artocarpus altilis (breadfruit) leaves across various fermentation durations (0, 7, 10, and 14 days) and to elucidate the molecular interactions between its key phytochemicals and essential antioxidant-regulating proteins. Methodology: Kombucha was produced under standardized conditions after performing phytochemical screening and pharmacognostic evaluation of the dried leaf powder. The antioxidant capacity was assessed using three complementary in vitro assays: DPPH, ABTS, and FRAP. Additionally, in silico molecular docking analysis was conducted to determine the binding affinities of principal phytochemicals (quercetin, kaempferol, gallic acid, morusin, and rutin) with major antioxidant-regulating targets, including Keap1, Nrf2, and PPARγ. Key Results: Antioxidant activity progressively enhanced with fermentation time, with the 14-day sample exhibiting the highest potency (IC₅₀ values: 26.31 ppm for DPPH, 41.46 ppm for ABTS, and 62.46 ppm for FRAP), which is comparable to ascorbic acid. Phytochemical profiling identified flavonoids, tannins, and terpenoids as the primary bioactive constituents. Molecular docking results showed strong binding affinities between the identified compounds and Keap1, Nrf2, and PPARγ, supporting a multi-target mechanistic role in mitigating oxidative stress. Conclusions: A. altilis leaf kombucha serves as a potent multifunctional antioxidant beverage that effectively integrates traditional herbal medicine with modern fermentation science and computational validation. The fermentation process significantly boosts the bioactive potential of the substrate. Recommendations: A 7-day fermentation period is recommended for commercial products to balance antioxidant benefits with sensory acceptability, while a 14-day period is ideal for maximum potency. Future research should include in vivo validation and molecular dynamics simulations to further verify these mechanistic insights.

Objective: This study aimed to investigate the antioxidant properties of kombucha prepared from Artocarpus altilis (breadfruit) leaves across various fermentation durations (0, 7, 10, and 14 days) and to elucidate the molecular interactions between its key phytochemicals and essential antioxidant-regulating proteins. Methodology: Kombucha was produced under standardized conditions after performing phytochemical screening and pharmacognostic evaluation of the dried leaf powder. The antioxidant capacity was assessed using three complementary in vitro assays: DPPH, ABTS, and FRAP. Additionally, in silico molecular docking analysis was conducted to determine the binding affinities of principal phytochemicals (quercetin, kaempferol, gallic acid, morusin, and rutin) with major antioxidant-regulating targets, including Keap1, Nrf2, and PPARγ. Key Results: Antioxidant activity progressively enhanced with fermentation time, with the 14-day sample exhibiting the highest potency (IC₅₀ values: 26.31 ppm for DPPH, 41.46 ppm for ABTS, and 62.46 ppm for FRAP), which is comparable to ascorbic acid. Phytochemical profiling identified flavonoids, tannins, and terpenoids as the primary bioactive constituents. Molecular docking results showed strong binding affinities between the identified compounds and Keap1, Nrf2, and PPARγ, supporting a multi-target mechanistic role in mitigating oxidative stress. Conclusions: A. altilis leaf kombucha serves as a potent multifunctional antioxidant beverage that effectively integrates traditional herbal medicine with modern fermentation science and computational validation. The fermentation process significantly boosts the bioactive potential of the substrate. Recommendations: A 7-day fermentation period is recommended for commercial products to balance antioxidant benefits with sensory acceptability, while a 14-day period is ideal for maximum potency. Future research should include in vivo validation and molecular dynamics simulations to further verify these mechanistic insights.