Sophorolipid was produced by Candida bombicola ATCC 22214 from soybean dark oil, a byproduct of soybean oil processing. With a fed-batch culture of C. bombicola for 7 days, 90 g/l of sophorolipid was obtained. The CMC (critical micelle concentration) and minimum surface tension of the sophorolipid in aqueous solution were found to be 150 mg/l and 48 mN/m, respectively. The dispersion capability of sophorolipid was higher than that of the chemical surfactants such as SDS and Brij30. The molar solubility ratio (MSR) of 4-methylnaphthalene was 0.2. Linoleic and oleic acids were the main constituents of the fatty acid composition of the sophorolipid. The sophorolipid showed antimicrobial activity against Propionibacterium acne and Bacillus subtilis.