Environmental Regulation, Molecular Profiling, and Preliminary Functional Evaluation of Extracellular Vesicles from Pleurotus tuber-regium

particles/mL at 34 °C, approximately six-fold higher than at 30 °C. Transcriptomic profiling showed coordinated repression of cell wall-associated genes and redox enzymes, together with induction of endoplasmic reticulum proteostasis pathways, consistent with stress-associated changes in the cellular context of vesicle release. Ultrasonicated PTR-EVs exhibited enhanced DPPH and ABTS radical-scavenging activities in chemical assays, with DPPH increasing from 59.52% to 71.73% and ABTS from 38.25% to 40.51%. Encapsulation efficiencies reached 32.67% ± 1.3% for proanthocyanidins and 46.01% ± 0.5% for curcumin. PTR-EVs showed the best short-term stability at pH 7 and 4 °C, supporting their further evaluation as an edible fungal vesicle platform for food-related nanoscale delivery.