Endophytic Berkeleyomyces rouxiae producing podophyllotoxin and deoxypodophyllotoxin with antibacterial, antioxidant, and cytotoxic activities

Background Podophyllum hexandrum is a medicinal plant valued for podophyllotoxin and deoxypodophyllotoxin, whose sustainable production remains challenging. Endophytic fungi represent potential alternative sources of these bioactive metabolites. Methods Endophytic fungi isolated from P. hexandrum were screened using thin-layer and high-performance liquid chromatography. The promising isolate was characterized using morphological, molecular, chromatographic, and spectroscopic analyses. The antibacterial, antioxidant, and cytotoxic activities of the fungal-derived compounds were evaluated. Results Among 25 isolates, PH12 was identified as Berkeleyomyces rouxiae, producing podophyllotoxin (53.8 µg/g DW) and deoxypodophyllotoxin (32.3 µg/g DW), with structural analyses confirming metabolite identity. Fungal podophyllotoxin (fPT) and deoxypodophyllotoxin (fDPT) showed IC50 values of 2.491 ± 0.08 and 2.602 ± 0.09 µg/mL against Escherichia coli, and 2.388 ± 0.07 and 2.528 ± 0.07 µg/mL against Staphylococcus aureus, respectively, compared with 2.467 ± 0.08 and 2.519 ± 0.09 µg/mL for their standards against E. coli and 2.243 ± 0.06 and 2.459 ± 0.08 µg/mL against S. aureus. The fPT and fDPT demonstrated concentration-dependent antioxidant activity, with IC50 values of 325.6 and 404.2 µg/mL in the DPPH assay and 339.3 and 465.1 µg/mL in the reducing power assay, compared with 302.7 and 370.9 µg/mL and 306.6 and 439.1 µg/mL for standards. Cytotoxicity against MCF-7 cells showed IC50 values of 1.336 µg/mL for fPT and 18.32 ng/mL for fDPT, comparable to 1.284 µg/mL and 16.46 ng/mL for standards. Conclusion This is among the first reports of B. rouxiae producing both lignans, highlighting its potential as a sustainable source of pharmacologically important compounds with antibacterial, antioxidant, and cytotoxic activities.