In an ongoing survey for bioactive microbial metabolites from different biospheres of India, a new marine bacterium identified as Pontibacter korlensis strain SBK-47 was isolated from the coastal waters of Karaikal, Puducherry, India, which produced a novel lipopeptide biosurfactant. The biosurfactant was purified and structurally elucidated as Palmitic acid-Ser-Asp-Val-Ser-Ser based on TLC, FT-IR, NMR, GC-MS, HPLC, MALDI-TOF and tandem MS analysis. This novel lipopeptide biosurfactant was named as Pontifactin. Pontifactin exhibited a surface tension reduction and critical micelle concentration (CMC) of 25 mN/m and 25 mg/L, respectively. Furthermore, the biosurfactant showed emulsification and surface tension stability over a wide range of pH (4-10) and temperature up to 100°C. Pontifactin showed promising antimicrobial activity against Streptococcus mutans, Micrococcus luteus, Salmonella typhi and Klebsiella oxytoca at a biosurfactant concentration ranging between 1-2 mg/mL and maximum antibiofilm activity at the biosurfactant concentration of 2 mg/mL against Bacillus subtilis, Staphylococcus aureus, Salmonella typhi and Vibrio cholerae. This is the first report on Pontifactin, a multifunctional lipopeptide biosurfactant, produced by a marine Pontibacter korlensis strain SBK-47, exhibiting promising surface-active, antimicrobial and anti-biofilm activities and thus finds possible use in biomedical applications.