Biosurfactant is a biodegradation accelerator that improves bioavailability and facilitates degradation by microorganisms. The study was meant to produce a novel biosurfactant molecule from Bacillussafensis YKS2. An efficient biosurfactant-producing strain, namely, Bacillus safensis YKS2, was selected using hemolytic activity, drop collapsing test, oil spreading test and blue agar plate methods in four oil-degrading strains isolated from a soil sample. Biosurfactant production in the optimization of bacteria culture conditions by RSM is a statistical grouping technique that is analyzed using the AVOVA approach to surface tention. In addition, the study was characterized by UV spectrophotometer FT-IR, HR-SEM, and GC-MS analyses to explain its structural and chemical details. Wastewater treatment was monitored for pH, EC, turbidity, alkalinity, chemical oxygen demand (COD), biochemical oxygen demand (BOD) and dissolved oxygen (DO) in order to justify the efficacy of the biosurfactant during wastewater treatment. The results of the UV spectrophotometer showed absorption at 530 nm, and the FT-IR analyzed carboxylic acids, alcohol and phenols groups, whichthe GC-MS analysis indicated were lipopeptide purified by hexadecanoic andtetradecanoic processes, respectively. The results show that the wastewater removal efficiency of 70% wasachieved within 24 h. In comparison, metagenomics was conducted during the treatment process to identify changes in the microbial load and diversity, which essentially indicatethe biosurfactant performance of the wastewater treatment process. The microbial load in the treated biosurfactant wastewater (84,374 sequences) was greatly decreased compared to untreated wastewater (139,568 sequences). It was concluded that B. safensis YKS2, producing a glycolipid form of biosurfactant, has possible benefits in the remediation of wastewater, and can be used for large-scale processing inbiosurfactant industries.