Silver nanoparticles have been synthesized by several methods, but each of them has disadvantages. Chemical methods, have high toxicity and low stability, and physical methods, are costly and low-efficient. More recently, researchers have found a new technique for synthesizing AgNPs, using plants and microorganisms such as bacteria, fungi, and Actinomycetes. This method is called “eco-friendly”, “green synthesis” or “Biosynthesis”. The aim of this investigation was, biosynthesis of AgNPs using Saccharomyces cerevisiae, which is one of the safe microorganisms that the AgNPs synthesized by it, are suitable for using in pharmacology and medicine purposes. Saccharomyces cerevisiae was cultured in sabraud dextrose broth and incubated at 28-30oc for 24 h, then it was centrifuged, and its supernatant was added to the silver nitrate solution with the concentration of 0.001, 0.002 and 0.003 M at pH: 5.6, 7 and 9, then incubated at 28 -35 and 37°c. The formation of AgNPs, was monitored by color-changing, uv-vis spectroscopy, TEM, XRD and FTIR. Antifungal effect of AgNPs on Microsporumcanis investigated by disc diffusion method. MIC and MFC were also measured. The results, showed that Saccharomyces cerevisiae have had the great potential for synthesizing of AgNPs in the size of 27 nm and have shown the antifungal effect in the concentration of 33 ± 0.29 m g/ml. In this study, we have reported a simple, biological, easy and nontoxic method for synthesis of silver nanoparticles, which have had suitable antifungal effect on Microsporumcanis, by using one of the nonpathogenic microorganisms.