The diacetylated lactonic sophorolipid from Candida bombolica was converted into the deacetylated acidic form by alkaline hydrolysis and subsequently treated with several glycosidases. One of these enzymes, a hesperidinase (E.C. 3.2.1.40), was most active in the specific release of one glucose molecule from the disaccharide lipid. The novel glucolipid was isolated and characterized. The surface and interfacial tension of aqueous solutions were measured and compared with the lactonic and acidic sophorolipid.