Background: It is generally known that several lactic acid bacteria (LAB) species have antibacterial properties. Different Lactobacillus species have been shown to have anticandidal effects by numerous researchers. This genus's members benefit the vagina and gastrointestinal systems' health. Additionally, they create a variety of bacteriocins with a wide spectrum of actions against bacteria or fungi. Objective: The aim of the current study was to examine the antifungal effects of bacteriocin produced by Lactobacillus spp. isolates against Candida spp. with the ultimate goal of gene expression on C. albicans biofilm genes before and after bacteriocin treatments. Material and methods: To test the antifungal activity of lactobacilli that make bacteriocin against the yeast Candida, 50 yogurt and vaginal samples were gathered. Testing on the isolated Lactobacilli included microscopic, macroscopic, and biochemical examinations. Additionally, a screening was done to find the best producer isolation. A total of 100 urine samples from females of various ages were obtained for the isolation of Candida spp. All of the isolated C. albicans were examined under a microscope, on a larger scale, and using biochemistry. Primary and secondary screening methods were used to look into the detection of bacteriocin formation from Lactobacillus. The expression of the agglutinin-like sequence 1 (ALS1) gene before and after treatment with bacteriocin was compared using reverse transcription quantitative polymerase chain reaction (RT-qPCR). Results: The agar wells diffusion technique was the most effective, according to the results. The results show that folding of this gene was reduced after bacteriocin treatment which means that bacteriocin inhibits the gene expression of biofilm formation of C. albicans. Conclusions: Our findings suggest that bacteriocin treatments may offer an effective substitute for antifungal medications. To verify these results, additional in vivo research is required.