Oral cavity is an ideal place for endogenous microbes to colonize as biofilm on the dentures causing oral infection. The study comprises denture wearers and non–denture wearers. The prevalence of Lactobacillus and Klebsiella Spp from denture flora of test and palatal flora of control groups was studied correlating the outcome with food, hygiene and age of the subjects. Samples were collected, cultured on to specific media and identified by standard methods. Data was statistically analyzed using student’s t and “chi–square test”. The result was correlated with the subject’s denture/oral profile and microbial prevalence. In both the groups Lactobacillus spp. (78%) were the predominant and diverse bacterial isolates than Klebsiella Spp.(38%). Six Lactobacillus spp. were isolated from the test and four in control group. K. pneumonia and K. oxytoca were the only species in both the groups. Correlation was found between the higher prevalence of Lactobacillus than Klebsiella spp. and their lifestyle.