Unveiling bioactive components of Gymnema sylvestre for antimycotic potential and process optimization.

Phyto-compounds play a considerable role in alleviating fungal diseases. However, the process for extraction, optimization, validation, comprehensive characterization and thorough evaluation is imperative to get the lead molecule. Gymnema sylvestre is loaded with multiple bioactives, therefore, the ultrasound-assisted extraction (UAE) process was optimized to produce bioactive components fraction (EGS), highly effective to inhibit storage pathogens, exhibiting significant effect against Aspergillus parasiticus 6365 (EC50 97.2 µg/mL) and Penicillium expansum 2995 (EC50 117.9 μg/mL). To ensure bulk production feasibility, the process was optimized using a factorial design, revealing a time of 17.45 min, 30.65 kHz. amplitude and 38.50 mL solvent to biomass ratio for the maximum yield (365.51 mg/ g sample). An untargeted metabolomics analysis unveiled the appearance of fifty-one compounds with the abundance of gymnemic acid I-VII, gymnemanol, gymnemagenin, gymnemoside C-E, gymnemasin A, D. Molecular modelling displayed the highest contribution of gymnemanol (- 20.00 kJ/mol), gymnemic acid II (- 19.6 kJ/mol) and gymnemagenin (- 19.5 kJ/mol) to inhibit the target-specific site of the fungal protein. Fungal growth inhibition was explained through molecular interactions, suggesting stable protein-triterpene complex formation with low distance H-bonds, hydrophobic pi-alkyl bonds. Major components predicted to exhibit a strong binding mechanism with the selective amino acid residues could be responsible for antimycotic action. Therefore, these findings provide empirical support to further explore these bioactive compounds as a promising, eco-friendly antimycotic agent. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-025-02020-9.