A biosurfactant produced by Tsukamurella sp. 26A was purified by procedures including acid precipitation, ethylacetate extraction, and adsorption chromatography. The purified biosurfactant reduced the surface tension of water from 72 mN/m to 30 mN/m at a concentration of 250 mg/l, whereas the minimum interfacial tension against n-hexadecane was lowered to 1.5 mN/m at a concentration of 40 mg/i. The compound stabilized oil-in-water emulsions with a variety of commercial oils and had strong emulsification and stabilization activities when compared to those of commercial emulsifiers and stabilizers. Surface tension was stable over a broad range of pH (2-12) and temperature ($100^{circ}C$, 3h). The biosurfactant was identified as glycolipid having a hydrophilic moiety of trehalose.