Ethylene glycol is an environmental pollutant, which exists in airport runoff and industrial waste. In this article, biodegradation of ethylene glycol in a two-chamber, batch-mode microbial fuel cell was investigated. Glucose and ethylene glycol at different concentrations were used as carbon and energy sources. Chemical oxygen demand removal in the range of 92–98% indicated that microbial fuel cell can be used for biodegradation of ethylene glycol. Ethylene glycol also improved power generation and the maximum power density was 5.72 mW/m2 (137.32 mW/m3), with respect to the same glucose and ethylene glycol concentrations (500 ppm).