This study focuses on the development of an air-lift bio-electrochemical reactor (ALBER) with a continuous feeding regime. The objective is to enhance nitrogen removal from synthetic wastewater with a low carbon-to-nitrogen (C/N) ratio. The chemical oxygen demand (COD) and total nitrogen (TN) of the influent wastewater were 500 and 200 mg/L, respectively. The effect of four independent variables, i.e., temperature, hydraulic retention time (HRT), N−NH₄⁺/TN ratio and current density in the range of 16–32 °C, 6–12 h, 25–75%, and 2–10 A/m², respectively, at three levels on the bio–electrochemical reactor performance were investigated during the bio–electrochemical reactor operation. The Face Center Cube (FCC) of response surface methodology (RSM) was used for design of experiments and model of obtained data. The ALBER achieved the maximum TN removal of 73% (146 mg/l) using external voltage and zeolite/plastic medium at temperature of 16 °C, HRT of 6 h, current density of 2 A/m² and N−NH₄⁺/TN ratio of 75%. The results indicated that shortening the HRT from 12 to 6 h, reducing the temperature from 32 °C to 24 °C, increasing the current density from 2 to 6 A/m² and the reduction of nitrate concentration caused an increase in the TN removal. The results indicated that the performance of air-lift bio-electrochemical for nitrogen removal could be attributed to autotrophic denitrification (AD) and simultaneous nitrification/denitrification (SND). The research findings suggest that the ALBER should be further studied for potential use in treating industrial wastewater at low temperatures.