In order to decrease the influence of acid rain and air pollution caused by sulfur dioxide released from the combustion of fuel oils, more and more regulations on sulfur content in fuel oils are being implemented. The current industrial method for removing sulfur from fuels is hydrodesulfurization (HDS). HDS is costly and energy-intensive for deep desulfurization, requiring high temperature and high pressure. Furthermore, HDS is not effective for removing heterocyclic sulfur compounds such as Dibenzothiophene (DBT) and DBT derivatives. Biodesulfurization (BDS) is a process that removes sulfur from fossil fuels using a series of enzyme-catalyzed reactions. It leads to the development of highly efficient reactions and environmentally friendly technologies. UP-3 (CGMCC No.0973), which was isolated from soil and sewage contaminated by sulfur-containing petroleum in the Shengli Oilfield, was investigated in this paper. The species was identified by measuring biochemical characteristics and the DNA sequence of the 16S-rDNA gene. The results indicate that UP-3 belongs to Agrobacterium tumefaciens. UP-3 has the industrialized potential based on undegrading n-dodecane, n-hexadecane, paraffine and naphthalin. Optimized cultivation conditions of UP-3 were as follows: the growth time was 30h, the growth temperature was 31ºC, and the initial pH value was 7.3-7.5. Under optimal cultivation conditions, the 1200mg/L Dibenzothiophene (DBT) content was reduced by 87.5% using growth cells of UP-3 within 50h in aqueous system, and the rate of sulfur removal was about 70% of 1150mg/L DBT in model oil system when ratio of oil to water was 1:10.