In order to develop novel glycolipid biosurfactants, Pseudozyma parantarctica JCM 11752(T), which is known as a producer of mannosylerythritol lipids (MEL), was cultivated using different sugar alcohols with the presence of vegetable oil. When cultivated in a medium containing 4 % (w/v) olive oil and 4 % D-ribitol or D-arabitol, the yeast strain provided different glycolipids, compared to the case of no sugar alcohol. On TLC, both of the extracted glycolipid fractions gave two major spots corresponding to MEL-A (di-acetylated MEL) and MEL-B (mono-acetylated MEL). Based on (1)H NMR analysis, one glycolipid was identified as MEL-A, but the other was not MEL-B. On high-performance liquid chromatography after acid hydrolysis, the unknown glycolipid from the D-ribitol culture provided mainly two peaks identical to D-mannose and D-ribitol, and the other unknown glycolipid from the D-arabitol culture did two peaks identical to D-mannose and D-arabitol. Accordingly, the two unknown glycolipids were identified as mannosylribitol lipid (MRL) and mannosylarabitol lipid (MAL), respectively. The observed critical micelle concentration (CMC) and surface tension at CMC of MRL were 1.6 × 10(-6) M and 23.7 mN/m, and those of MAL were 1.5 × 10(-6) M and 24.2 mN/m, respectively. These surface-tension-lowering activities were significantly higher compared to conventional MEL. Furthermore, on a water-penetration scan, MRL and MAL efficiently formed not only the lamella phase (L(α)) but also the myelins at a wide range of concentrations, indicating their excellent self-assembling properties and high hydrophilicity. The present two glycolipids should thus facilitate the application of biosurfactants as new functional materials.