Fifty hydrocarbon-metabolizing microorganisms were isolated from soil samples polluted by the petroleum oils in Gamman-dong, Busan. Among them, strain 2-3A, showing strong emulsification activity, was selected by oil film-collapsing method. This bacterium was identified as Acinetobacter sp. and designated as Acinetobacter sp. 2-3A. The optimum temperature and pH on the growth of Acinetobacter sp. 2-3A were 25℃ and pH 7.0, respectively. The carbon and nitrogen sources for the most effective emulsification activity were 3.0% olive oil and 0.5% peptone, respectively. The 0.15% potassium phosphate was the most effective emulsification activity as a phosphate source. The optimum emulsification activity condition was 20℃, pH 7.0, and 2.0% NaCl. The optimum time for the best production of biosurfactant was 27 hrs. The emulsification stability was maintained at the temperature range from 4℃ to 100℃, pH range from 6.0 to 10.0, and NaCl range from 0% to 10%. For the oil resolvability of the biosurfactant, the residual oils were investigated by gas chromatography. As a result, it was verified that the biosurfactant decreased and decomposed crude oils from nC10 to nC32.