The primary objective of this study was to assess the probiotic attributes and antifungal activity of lactic acid bacteria (LAB) against the fungus,Trichophyton tonsurans. Among the 20 isolates screened for their antifungal attributes, isolate MYSN7 showed strong antifungal activity and was selected for further analysis. The isolate MYSN7 exhibited potential probiotic characteristics, having 75 and 70% survival percentages in pH3 and pH2, respectively, 68.73% tolerance to bile, a moderate cell surface hydrophobicity of 48.87%, and an auto-aggregation percentage of 80.62%. The cell-free supernatant (CFS) of MYSN7 also showed effective antibacterial activity against common pathogens. Furthermore, the isolate MYSN7 was identified asLactiplantibacillus plantarumby 16S rRNA sequencing. BothL. plantarumMYSN7 and its CFS exhibited significant anti-Trichophytonactivity in which the biomass of the fungal pathogen was negligible after 14 days of incubation with the active cells of probiotic culture (106CFU/ml) and at 6% concentration of the CFS. In addition, the CFS inhibited the germination of conidia even after 72 h of incubation. The minimum inhibitory concentration of the lyophilized crude extract of the CFS was observed to be 8 mg/ml. Preliminary characterization of the CFS showed that the active component would be organic acids in nature responsible for antifungal activity. Organic acid profiling of the CFS using LC-MS revealed that it was a mixture of 11 different acids, and among these, succinic acid (9,793.60 μg/ml) and lactic acid (2,077.86 μg/ml) were predominant. Additionally, a scanning electron microscopic study revealed that CFS disrupted fungal hyphal structure significantly, which showed scanty branching and bulged terminus. The study indicates the potential ofL. plantarumMYSN7 and its CFS to control the growth ofT. tonsurans. Furthermore,in vivostudies need to be conducted to explore its possible applications on skin infections.