In this study, we devised a biofuel cell (BFC) by impregnating sheet-like cellulose nanofiber (CNF) with liquid fuel (fructose) and sandwiching it with the electrodes, making the structure simple and compact. CNF was considered as a suitable material for BFC because it is biocompatible, has a large specific surface area, and exhibits excellent properties as a catalyst and an adsorbent. In this BFC device, graphene-coated carbon fiber woven cloth (GCFC) was used as the material for preparing the electrodes, and the amount of enzyme modification on the surface of each electrode was enhanced. Further, as the distance between the electrodes was same as the thickness of the sheet-shaped CNF, it facilitated the exchange of protons between the electrodes. Moreover, the cathode, which requires an oxidation reaction, was exposed to the atmosphere to enhance the oxygen uptake. The maximum power density of the CNF-type BFC was recorded as 114.5 µW/cm2 at a voltage of 293 mV. This is more than 1.5 times higher than that of the liquid-fuel-type BFC. When measured after 24 h, the maximum power density was recorded as 44.9 µW/cm2 at 236 mV, and the output was maintained at 39% of that observed at the beginning of the measurement. However, it is not the case with general BFCs, where the power generation after 24 h is less than 5%. Therefore, the CNF-type BFCs have a longer lifespan and are fuel efficient.