Microbial sources are regularly used as reliable biocatalysts sources which are often used in the process and production industry. Demands for such organisms with greater capacity of intended enzyme production are on the rise. Lipase is important enzyme used in the biotechnological process of hydrolysis of fats in almost all the relevant industries We have utilized the local oil-contaminated soil resources to search for efficacious bacterial strains that have excellent lipase activity. We were successful in identifying two such bacterial sources, namely, Bacillus subtilis strain RCPS3 and Bacillus fumarioli strain RCPS4, responsible for lipase production from oil effluent contaminated soil of Telangana. This is the first report of these two strains from this part of India that are involved in lipase production. The strains were isolated, optimized, and purified using standard microbiology protocols and were characterized at the molecular level using the biomarker 16s ribosomal RNA genes of the strains. The identified and isolated bacterial strains were confirmed as Bacillus subtilis strain RCPS3, and Bacillus fumarioli strain RCPS4 through molecular and computational characterization.