Optimization of mycosynthesis of silver nanoparticles using Inocutis levis and evaluation of their physicochemical and biological properties

Silver nanoparticles (AgNPs) were biosynthesized using the aqueous extract of Inocutis levis fruiting bodies and optimized by response surface methodology using a Box–Behnken design. Preliminary screening identified the 50% aqueous extract as the most suitable for AgNP synthesis, while the model-predicted optimum conditions were 7 mM AgNO₃, pH 8, and 46 °C. The biosynthesized AgNPs were characterized by UV–Vis, FTIR, FESEM, DLS, EDX, XRD, and zeta potential analyses, confirming predominantly spherical and crystalline nanoparticles. FESEM showed an average particle size of 18.2 ± 4.5 nm, while DLS revealed a hydrodynamic diameter of 92 nm and a zeta potential of − 37.7 mV. The biosynthesized AgNPs exhibited antioxidant activity (ABTS IC₅₀ = 3.42 mg/mL) and antibacterial activity against Gram-positive and Gram-negative bacteria (MIC = 128–512 µg/mL), while inhibiting biofilm biomass formation by 67.39–94.93% at MIC. MTT analysis demonstrated > 80% cell viability in human dermal fibroblasts at 20–60 µg/mL, indicating limited cytotoxic effects under the tested conditions. Overall, these findings demonstrate the potential of I. levis as a fungal platform for the sustainable biosynthesis of silver nanoparticles with favorable physicochemical characteristics and biological activities, highlighting their potential for future biomedical applications.