Rhodococcus sp 51T7 produced a trehalose 2,3,4,2’ tetraester with surface active properties. When grown on hydrocarbon, cells were highly segmented and accumulated lipid granules in the cytoplasm. Production and glycolipid composition was affected by the nature of the carbon source. Optimal concentrations of sodium nitrate, potassium phosphate and iron were : 2.5, 1.5 and 0.01 g/L respectively. Surfactant production is growth-associated. Production was increased from 0.5 g/L to 3 g/L of glycolipid.