The effect of cerulenin, anti-lipogenic antibiotic, on the production and fatty-acid profiles of biosurfactants (mannosyl-erythritol lipids) of Candida antarctica were investigated by changing the chain-length of fatty acid as the carbon source. On longer-chain acid (C14 to C18), the production and fatty-acid profiles of the biosurfactants were not entirely affected by the antibiotic, indicating that the yeast is able to synthesize the biosurfactants from those substrates without the operation of “de novo synthesis system” of fatty acid. These results thus confirm our previous presumption that a new type of “chain-shortening system” (partial β-oxidation system) mainly contributes to the formation of the extracellular glycolipids as biosurfactants.