Biosurfactants are surface-active compounds produced by microorganisms. These molecules reduce surface tension between aqueous solutions and hydrocarbon mixtures. In this study, we collected water samples from Jeppiaar salt pan in Chennai, Tamilnadu, India. Ten different microorganisms were isolated from salt pan water samples. Screening of biosurfactants producing potential strain was done using Hemolytic activity, Oil displacement test, Drop Collapse assay. Four isolates were positive for biosuractant production among that one isolate was found as potential biosurfactant producer. Both biochemical and 16S rRNA sequencing methods one of the potential strain was identified as Bacillus cereus JS06. The biosurfactant was extracted from cell-free broth at 72-h grown cells by step-by-step purification of acid precipitation method. The dried surfactant was used for further characterization by Fourier transmission Infrared spectroscopy with attenuated total reflectance (FTIR-ATR) and anticancer studies. On FTIR-ATR analysis characteristic absorbance bands were observed for the extracted biosurfactant. The crude biosurfactant was screened for its anticancer activity against human laryngeal carcinoma (HEP-2) cell line and transformed aneuploid immortal keratinocyte (HeCAT) cell line. The results obtained showed that biosurfactants have better interfacial-activity and anticancer property, thus being more attractive to be applied in pharmaceuticals and medical field.