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• Catalytic properties of Ni-Fe and GF/Pt were characterized using LSV tests. • Tafel slope was determined based on the data points in the onset potential. • EGF can be used as substrate for hydrogen production. • Ni-Fe shows comparable performance with GF/Pt cathode in BES application. The high cost of Pt-based cathode, and its possibility of being poisoned by the presence of buffer in the electrolyte are two paramount issues in the BES system. The Ni-Fe has become one of the good alternatives because it has excellent catalytic properties, inexpensive, commercially available and low toxicity to microorganisms. In this study, Ni-Fe foam applied as a cathode in dual-chamber BES, while effluent of glucose fermentation as a substrate in the anode side. The characteristic of Ni-Fe surface was analyzed by using field emission scanning electron microscopy. Whereas, the catalytic property of Ni-Fe was evaluated by using linear sweep voltammetry test. The maximum hydrogen production rate and yield obtained were 500 ± 80 m3/m3/d and 470.2 ± 11.2 mL/g COD, respectively. The results show that the Ni-Fe has comparable performance to GF/Pt. Hence it could be used as an alternative cathode in BES application. [ABSTRACT FROM AUTHOR]