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The objective of the present study was to isolate the biosurfactant (BS) producing lactic acid bacteria (LAB) from traditional fermented food (buttermilk) and its functional and structural characterization. BS isolated from strain MRTL9 reduced surface tension from 72.0 to 40.2 mN m^-1. The critical micelle concentration (CMC) of BS was 2.25 mg ml^-1 with emulsification efficiency (E₂₄) after 24 h of 64% against kerosene oil. The cell bound BS was partially purified by silica gel column chromatography and found as glycolipid. The gas chromatography and mass spectroscopy data revealed the fatty acid as hexadecanoic acid. Xylose was determined as hydrophilic moiety. The BS was found to be stable to pH changes over a range of 4.0-12.0, being most effective at pH 7 and showed no apparent loss of surface tension and emulsification efficiency after heat treatment at 120°C for 15 min. The outcomes of cellular …