Shiitake () is an Asian edible mushroom with the second-largest cultivation percentages among edible mushrooms in the world. Previous studies have shown its nutritional richness such as high quantities of proteins, minerals, and vitamins. Moreover, and animal studies have displayed the medical importance of shiitake extracts including antitumor, antiviral, antibiotic, and hypocholesterolemic actions and also shown to have antibacterial efficacy and has been used for the elimination of oral biofilms and as a substitute of current chemical-based treatments. This study aims to analyze the inhibitory and antibacterial efficacy of shiitake mushroom extracts on periodontal pathogens.Double extract, i.e., hot water and ethanol extract of shiitake were used for the assessments of minimum inhibitory concentrations on , and , and using the tube-dilution method, and the time-kill curve was assessed for the above bacteria.Shiitake has shown potent antibacterial effects at a concentration of 3.12 μg/ml and has shown no growth after 2 h of culture.Shiitake mushroom extract has shown the potential antimicrobial effect on the certain periodontal pathogens.