Mycosynthesis of stable silver nanoparticles (AgNPs) using Aspergillus caespitosus exo‐ and endo‐filtrates were obtained. Both exo‐ and endo‐AgNPs showed an absorbance peak at 410 nm, spherical, hexagonal, and anisotropic crystalline nanoparticles with 22–57 nm in size. FTIR analysis of mycosynthesized AgNPs showed presence of amide and carbonyl functional groups, which depicts the presence of peptides that involved in the synthesis and stability of AgNPs. Nitrate reductase activity was found to be 0.71 and 0.36 μM/min/ml for exo‐ and endo‐filtrates, respectively. The optimum conditions for the synthesis of exo‐ and endo‐AgNPs were at 6:4 and 8:2 filtrate:double distilled water dilution, respectively. Also, both AgNPs syntheses were the maximum at using one mM AgNO3 in aqueous solution, 24‐h incubation at 30°C under static and illuminating conditions. Aspergillus caespitosus‐synthesized AgNPs showed antimicrobial activities against Gram‐positive and Gram‐negative bacteria and Candida albicans. They showed non‐cytotoxic effect with exception of endo‐AgNPs on MCF‐7 breast carcinoma cells. Different textile samples treated with these nanoparticles exhibited antimicrobial activities against the tested pathogenic microorganisms. Finally, Aspergillus caespitosus is a promising nano‐factory for simple, green, and cost‐effective biosynthesis of AgNPs using extracellular and intracellular filtrates.