The development of a convenient and sensitive sensor such as a microbial fuel cell (MFC) to monitor the operation of upflow anaerobic sludge blanket (UASB) is indispensable. However, the biosensor's properties were affected due to excessive acidification and suffocation of the electron transport. In this study, zero-valent iron (ZVI) was applied to restrain excessive acidification and improve the sensing performance. According to the results, the response rate of electrical signal accumulated with the addition of ZVI compared to the control reactor. As well as the electrical signal amplified and the subsidence rate maximum reached 0.059 V/h with 30 mg/L ZVI added that 883% higher than the control one during the stage (COD concentration 500 mg/L-1000 mg/L). With the electrochemical analysis, the internal resistance of ZVI-UASB-MFC decreased and redox activity promoted effectively with ZVI added. During the overloading phase, the fractional content of butyric acid changed from 53% to 31%, while that of acetic acid rose from 18% to 39% after 30 mg/L ZVI addition. These results indicated that adding ZVI to the digestion could retard excessive acidification by promoting butyric acid conversion and accumulating direct interspecies electron transfer simultaneous for enhancing the biosensor's performance. According to the Fe2+ and Fe3+ of effluent were 2.25 mg/L and 0.39 mg/L with 50 mg/L ZVI addition, moderate amount of ZVI was effective for system and safety to the environment. It might helpfully provide a promising way to enhance biosensing. (C) 2017 Elsevier B.V. All rights reserved.