The aim of this study was to isolate and characterize marine bacterial strains capable of converting selenite to elemental selenium with the formation of Se nanoparticles (SeNPs). For the first time, a novel marine strain belonging to Bacillus amyloliquefaciens (GenBank accession no. MK392020) was isolated from the coast of the Caspian Sea and characterized based on its ability for transformation of selenite to SeNPs under aerobic conditions. The preliminary formation of SeNPs was confirmed via color changes and the products characterized by UV-Vis spectroscopy. The field-emission scanning electron microscopy (FESEM) together with energy-dispersive X-ray (EDX) analysis showed the presence of the spherical SeNPs on both the surface of the bacterial biomass and in the supernatant solution. Dynamic light scattering (DLS) analysis showed the SeNPs to have an average particle size (Z-average) around 45.4-68.3 nm. The X-ray diffraction (XRD) studies substantiated the amorphous nature of the biosynthesized SeNPs. Fourier-transform infrared spectroscopic (FTIR) studies of the SeNPs indicated typical proteinaceous and lipid-related bands as capping agents on the SeNPs. Different effective parameters corresponding the yield of SeNPs by B. amyloliquefaciens strain SRB04 were optimized under resting cell strategy. Results showed that the optimal process conditions for SeNP production were 2 mM of selenite oxyanion, 20 g/L of cell biomass, and 60 h reaction time. The synthesized SeNPs had a remarkable antibacterial activity on Staphylococcus aureus compared with chloramphenicol as a broad-spectrum antibiotic.