The development of new signal systems for electrical biosensors could provide exciting new opportunities for biomedical analysis, pollutant monitoring, and explosive detection. The signal systems for commercial portable sensors involve the integration of a battery and a circuit conditioning system to power an amperometric biosensor. However, this increases the size and complexity of the entire system. In this study, we develop a simple amperometric biosensor that is directly powered by a button cell battery for the detection of lactate. A two-electrode sensing transducer was printed on cardboard or integrated on a ring. It was directly powered by a button cell battery, and connected to a multimeter for current measurement. This sensor showed a sensitive detection range of 0.04762–9.21429 mM and short measuring time of 2 min. These results show that this system can achieve an excellent sensing performance, and the construction of this new sensing system directly powered by a button cell battery offers a new method for further developing a wide range of miniaturized, flexible, portable, or wearable sensing systems, and these could be used in detecting various analytes that are important in medical diagnosis and environmental monitoring.