Silver nanoparticles (AgNPs) are nanomaterials obtained by nanotechnology and due to their antimicrobial properties have a major importance in the control of various species of bacteria, fungi and viruses, with applications in medicine, cosmetics or food industry. The goal of the paper was to present the results of the research carried out on rapid extracellular biosynthesis of silver nanoparticles mediated by culture filtrate of lactic acid bacteria Lactobacillus sp. strain LCM5 and to assess the antimicrobial activity. Analysis of transmission electron microscopy (TEM) micrographs evidenced that the size of AgNPs synthesized using culture filtrates of lactic acid bacteria strain LCM5 ranged between 3 and 35 nm diameter, with an average particle size of 13.84±4.56 nm. AgNPs presented a good dispersion, approximately spherical shape, with parallel stripes certifying crystal structure. Frequency distribution revealed that preponderant dimensions of biosynthesized AgNPs were below 20 nm (94%). Antimicrobial activity of AgNPs was variable depending on both species and group of test microorganisms (bacteria or fungi) involved. Diameter of growth inhibition zone of Aspergillus flavus and Aspergillus ochraceus caused by silver nanoparticles synthesized by lactic acid bacteria strain LCM5 were similar (12.39 ± 0.61mm and 12.86 ± 0.78 mm) but significant stronger inhibition was registered against Penicillium expansum (15.87 ± 1.01mm). The effectiveness of biosynthesized silver nanoparticles was more pronounced against Gram-negative bacteria Chromobacterium violaceum with larger zone of inhibition (18 ± 0.69 mm diameter) when compared to those from fungi. Results recommend the silver nanoparticles biosynthesized using culture filtrate of the lactic acid bacteria Lactobacillus sp. strain LCM5 for biotechnological purposes, as promising antimicrobial agents.