Exploiting suitable oxidation catalysts is of great importance in the development of sugar-based fuel cells (SFCs). Herein, a novel room-temperature glucose/O2 fuel cell (GFC), which employs 4-acetamido-2,2,6,6-tetramethylpiperidin-1-oxyl (ACT) as an anodic electrocatalyst and air-breathing Pt-C as a cathode, is demonstrated. Under room temperature operation, the as-assembled GFCs are capable of delivering a maximum power density of 100 μW cm-2 in the presence of 50 mM glucose. Bulk electrolysis products of glucose identified by mass spectrum and Fourier transform infrared spectroscopy include gluconic acid and glucaric acid, suggesting that the aldehyde and primary hydroxy groups of glucose can be deeply oxidized into carboxyl groups through a 6e- pathway. The deep glucose oxidation capability makes ACT a promising anodic electrocatalyst for SFCs.