Vegetable oil is the usual carbon source for the production of biosurfactants (BS), mannosylerythritol lipids (MEL). To simplify the procedures of BS production and recovery, we investigated the extracellular production of MEL from water-soluble carbon sources instead of vegetable oils by using two representative yeast strains. The formation of extracellular MEL from glucose was confirmed by thin layer chromatography (TLC) and HPLC analysis. On glucose cultivation, pure MEL were easily prepared by only solvent extraction of the culture medium, different from the case of soybean oil cultivation. The fatty acid profile of the major MEL produced from glucose was similar to that produced from soybean oil based on GC-MS analysis. The resting cells of Pseudozyma antarctica T-34 produced MEL by feeding of glucose only and gave a yield of 12 g l(-1). In contrast, Pseudozyma aphidis ATCC 32657 gave no MEL from glucose. Moreover, the extracellular lipase activities were detected at high levels during the cultivation regardless of the carbon sources. These results indicate that all the biosynthesis pathways for MEL in P. antarctica T-34 should constitutively function. In conclusion, P. antarctica T-34 thus has potential for BS production from glucose.