Green synthesis of silver nanoparticles (AgNPs) from G. stearothermophilus GF16: stable and versatile nanomaterials with antioxidant, antimicrobial, and catalytic properties.

Background: Silver nanoparticles (AgNPs) have attracted considerable interest for their distinctive physicochemical properties and wide-ranging applications in nanomedicine, environmental catalysis, and antimicrobial applications. However, sustainable and robust biosynthesis methods remain a challenge.

Results: In this study, we report the biosynthesis of thermostable AgNPs using the secretome of Geobacillus stearothermophilus GF16, a thermophilic and metal-resistant bacterium isolated from the hydrothermal volcanic area of Pisciarelli, Italy. The synthesis was performed without specialized growth media, relying solely on the cell-free bacterial supernatant, and was systematically optimized by varying precursor concentration, temperature, pH, and reaction time. The nanoparticles were characterized by UV-Vis spectroscopy, dynamic light scattering, Fourier-transform infrared spectroscopy, scanning (SEM) and transmission (TEM) electron microscopy. Morphological analysis showed predominantly subspherical nanoparticles with average diameters of 17 ± 5 nm (SEM) and 16 ± 5-7 nm (TEM), depending on precursor concentration. Thermogravimetric analysis demonstrated excellent thermal stability with retention of structural integrity up to 120 °C, an exceptional feature among biogenic AgNPs. The obtained AgNPs exhibited remarkable radical scavenging activity, reaching up to 79% in DPPH and 75% in ABTS assays at 100 µg/mL, highlighting a level of antioxidant performance rarely observed in AgNPs of bacterial origin. In addition to their redox properties, the nanoparticles demonstrated efficient catalytic activity as evidenced by the complete degradation of Congo Red in 20 min and 4-nitrophenol in 35 min. Time-kill assays and minimum inhibitory concentration (MIC) also showed a broad-spectrum antimicrobial potential with complete inhibition of Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella Typhimurium at 100 µg/mL. Interestingly, MIC values were significantly lower than those reported for comparable AgNPs. Notably, the nanoparticles also displayed hemocompatibility, validated by hemolysis assays performed on both healthy and β-thalassemic erythrocytes, with hemolysis rates consistently below the 2% safety threshold.

Conclusions: Overall, this study presents the first comprehensive characterization of AgNPs biosynthesized by a thermophilic bacterium, highlighting their multifunctional potential. The use of a thermophilic bacterium as a robust and flexible microbial nanofactory offers a novel eco-friendly and scalable strategy for AgNP production. The resulting nanoparticles exhibit unique thermal stability, broad-spectrum bioactivity, and clinically relevant hemocompatibility, underscoring their promising applicability in nanomedicine, green catalysis, and environmental remediation.