Seeking for simple, rapid, and environmental-friendly routes to produce metal nanoparticles is quite attractive for various biotechnological applications. Biological synthesis method of silver nanoparticles has been found very promising due to their non-toxicity and simplicity. Here, the spores of Bacillus stratosphericus isolated from soil enriched with 30 % H2O2 were used for the production of silver nanoparticles. Furthermore, the possible mechanism of silver nanoparticle synthesis by the spores was elucidated for the first time. In this regard, dipicolinic acid (DPA) was shown to play a critical role as a nanoparticle-producing agent. UV-Vis absorption spectroscopy, X-ray diffraction technique, energy-dispersive spectroscopy, and transmission electron microscopy were used to characterize the nanoparticles. Unlike vegetative cells of B. stratosphericus, the spores and the purified DPA were capable of producing nanoparticles from silver nitrate (AgNO3). These biogenic nanoparticles, which were highly toxic against different pathogenic bacteria, showed mixed structures including spherical, triangular, cubic, and hexagonal with the approximate size between 2 and 20 nm in diameter. Our results illustrated the role of dipicolinic acid as a main factor for the synthesis of nanoparticles by the bacterial spores.