The extractive membrane bioreactor (EMB) employs a dense silicone rubber membrane to selectively extract hydrophobic organic compounds from industrial wastewaters into a bioreactor in order to biodegrade them. The major drawback of the EMB is excess biofilm growth on the membrane, which limits mass transfer and creates oxygen limitations. In this work, nitrate has been used as an electron acceptor instead of oxygen. Due to the high solubility of nitrate in water, it is hypothesised that nitrate penetrates the whole biofilm, preventing the formation of inactive zones of bacteria. Four experiments have been performed with toluene as a model substrate under anoxic conditions. The effect of nitrate concentrations on the biofilm and on the toluene flux have been investigated. In addition, the production of soluble microbial products (SMPs), and bacterial hydrophobicity were studied. Under high nitrate concentrations, the performance of the anoxic EMB was stable and no biofilm was formed. The bacteria metabolised toluene, and the toluene flux remained approximately constant. Conversely, at low nitrate concentration, a decrease in pollutant flux concomitant with biofilm growth was observed. The production of SMPs increased under limiting nitrate concentrations, but the hydrophobicity of the suspended bacteria remained constant. However, the bacterial hydrophobicity of the attached cells was significantly greater than that of the suspended cells.