Conventional enzymatic biofuel cells (EBFCs) use glucose solution or glucose from human body. It is desirable to get glucose from a substance containing glucose because the glucose concentration can be kept at the optimum level. This work developed a biofuel cell that generates electricity from cellulose, which is the main components of plants, by using decomposing enzyme of cellulase. Cellulose nanofiber (CNF) was chosen for the ease of decomposability. It was confirmed by the cyclic voltammetry method that cellulase was effective against CNF. The maximum output of the optimized proposed method was 38.7 μW/cm2, which was 85% of the output by using the glucose solution at the optimized concentration.