Effectiveness of a microbial biosurfactant, sophorolipid, was evaluated in washing and biodegradation of model hydrocarbons and crude oil in soil. Thirty percent of 2-methylnaphthalene was effectively washed and solubilized with 10 g/L of sophorolipid with similar or higher efficiency than that of commercial surfactants. Addition of sophorolipid in soil increased biodegradation of model compounds: 2-methylnaphthalene (95% degradation in 2 days), hexadecane (97%, 6 days), and pristane (85%, 6 days). Also, effective biodegradation method of crude oil in soil was observed by the addition of sophorolipid, resulting in 80% biodegradation of saturates and 72% aromatics in 8 weeks. These results showed the potentials of the microbial biosurfactant, sophorolipid, as an effective surfactant for soil washing and as an in situ biodegradation enhancer.