Nowadays when control of environmental toxicity is a matter of concern, the focus of the researchers is to find an eco-friendly process. Considering the hazards associated with chemical synthesis of nanoparticles, green synthesis approaches have gained considerable attention for their sustainable nature in nanomedicine and nanobiotechnology. Here, exopolysaccharide (EPS) synthesized by a geothermal spring origin B. anthracis PFAB2 is used for green synthesis of silver nanoparticles (AgNPs). Elemental analysis of EPS-coated AgNPs exhibited solid peaks of silver (39.66%) along with oxygen and carbon. TEM analysis confirmed the hexagonal shape of the AgNPs. Polydispersity index (PDI) reinforced the moderately monodisperse nature of the nanoparticles. High negative zeta potential indicated longer shelf life, good colloidal nature and high dispersive nature of the AgNPs. The B. anthracis PFAB2 EPS-coated AgNPs demonstrated prospective biocidal characters for both gram positive and gram negative bacteria along with some potentially hazardous fungi compared with the conventional antimicrobials. The results can be much expedient in the future for treatment of microorganisms that are otherwise resistant to traditional antibiotics or antifungal drugs.