We fabricated a photo-driven bioanode on laser-induced graphene (LIG) using a facile one-step laser-scribed technique. Nb4C3Tx MXene was decorated on the LIG surface to induce a high surface area for immobilizing the photocatalyst. The thylakoid membrane extracted from Spinacia oleracea utilizes its intrinsic photosynthetic and catalytic capability for efficient energy conversion. A platinum-based gas diffusion electrode is used as the cathode material, allowing for charge transfer and oxygen reduction to produce water. The bioelectrochemical activities of the as-fabricated thylakoid-based biofuel cells were investigated. Upon illumination, the assembled biofuel cell exhibited an open circuit voltage of 0.45 V, a maximum photocurrent density of 68.69 µA/cm2, and a power density of 7.24 µW/cm2. The unique features of thylakoid membranes, Nb4C3Tx MXene, and the LIG substrate play a critical role in producing high-performance photo-biofuel cells. Our approach presents a promising strategy for harnessing the biochemical energy of plants to generate bioelectricity.