With the advantages of miniaturization, simple device structure, and fast response, the organic electrochemical transistor (OECT) has become an emerging platform for developing wearable enzyme sensors for real-time health monitoring. The floating gate (FG) OECT employs a distinct signal acquisition and amplification structure, mitigating the effects of non-specific physical adsorption during the sensing process and preventing contamination of the electrolyte solution by side reaction products. The current work reports a feasible wearable enzyme sensor using a poly(benzimidazobenzophenanthroline) (BBL)-Nafion-enzyme-Nafion stacking structure as the sensing layer of the FG OECT. Based on the experimental results, the BBL film with an area of 3.14 mm2 and a thickness of 175 nm can generate an open circuit potential of 199.61 mV in 10− 1 M hydrogen peroxide compared with the blank control. Then, the FG OECT is integrated with the flexible microfluidic systems for on-skin detection of glucose, lactate, and uric acid with sensitivities of 92.47, 152.15, and 74.27 µA·dec− 1, respectively. This FG OECT-based wearable enzyme sensor will open new windows for multiplexed detection of sweat metabolites.