Facile, efficient, and robust immobilization of metal nanostructures on porous bioscaffolds is an interesting topic in materials chemistry and heterogeneous catalysis. This paper reports an in situ method for the synthesis and immobilization of small silver nanoparticles (AgNPs) at room temperature on natural eggshell membrane (ESM). The eggshell membrane is the clear film lining the eggshell, visible when one peels a boiled egg, which presents interwoven fibrous structure and can be used as a unique protein-based biotemplate. Many chemical and physical techniques are available for the synthesis of antimicrobial silver nanoparticle (AgNPs), but the green synthesis is the most emerging method of synthesis. Here the silver nanoparticles were synthesized using Ocimum sanctum leaf extracts which act as both reducing and capping agent. Visual color change, UV spectroscopy, scanning electron microscopy and transmission electron microscopy unambiguously identifed the presence of AgNPs on ESM. Besides the antibacterial activity of these ESM’s was seen through the zone of inhibitions observed for the test samples against plates of Escherichia coli. Such egg membranes can be used against skin wounds like that in burns wherein the AgNPs can prevent the bacterial infections and the presence of ESM will also help the recovery of the skin as the amino acid composition of the egg membranes is quite close to that of human skin and other human tissues.