Uric acid (UA) is the primary waste product from purine metabolism in humans. Excessive UA levels in the body will accumulate in joints and form crystals that cause a wide range of health problems. An enzymatic electrochemical biosensor for UA based on the transition metal complex-incorporated polyaniline PANI-RC functionalized with both urate oxidase (UOx) as a specific bioreceptor and horseradish peroxidase (HRP) as a signal enhancer was developed. The transition metal complex being used herein is the commonly used redox couple (RC) in electrochemical biosensors, [Fe(CN)6]3-/4-, which plays the pivotal role of electron acceptors. This PANI-RC platform then becomes a conducive environment not only for enzyme immobilization but also for signal transfer improvement. The synergistic combination of HRP near UOx and RC anchored on the backbone of PANI helps in electron transfer from the enzymatic reaction to the current collector. The resulting PANI-RC-based UA sensor demonstrates high sensitivity with a detection limit of 11.4 μM, wide linear range, good stability, and excellent selectivity even in the presence of the most problematic interference in UA assays (e.g., ascorbic acid and urea). The recovery tests using artificial biofluid-spiked UA samples also showed promising results for practical usage of the PANI-RC-based UA sensor.