A large quantity of ethylene glycol(EG) is remained in the effluent after pretreating polyester weight-loss wastewater physicochemically in the fist stage and must be treated biologically in the second stage. Therefore, an excellent EG-utilizing bacteria strain was isolated from the natural system and the optimal culture conditions of the strain were investigated. The optimal culture conditions of temperature, pH, and nitrogen source were found to be 35$^{circ}C$, 7.5 and ammonium chloride, respectively, when CODCr removal efficiency was more than 90%. The growth of stains and EG removal efficiency was slightly improved by adding elements such as niacin and biotin. With increasing inoculation size in a batch culture, the removal efficiency of EG was conspicuously increased. Growth rate was inhibited when the initial concentration of EG was more then 30g/L. The strain was identified as Pseudomonas sp. based on morphological and biological characteristics and named as Pseudomonas sp. EG1.