Enzymatic biofuel cells are presented as an untethered alternative energy source that could power small implantable or wearable medical devices. However, most of these catalytic processes do not provide with enough energy to power common small electronic-mechanical devices. On the other hand, conducting polymer-based actuators are of great interest for their biocompatibility, flexibility, processability, possibility to be miniaturized and low power consumption. So far, these artificial muscles have been driven by external power sources that prevent them for being completely autonomous. There is a need for a novel power source to elaborate actuators that could use physiological processes as a driving force. These soft actuators’ low power consumption matches the electrical power generated by the biocatalysis of some enzymes, such as glucose oxidase and laccase in presence of glucose and oxygen in aqueous media. Here, we present the latest results in the development of polypyrrole-based soft actuators powered by enzymatic biofuel cells. The actuator consists of a tri-layer conductive substrate on which the polypyrrole is electrodeposited in both sides. The polypyrrole layers act as the active part, expanding and contracting upon a redox reaction, resulting in a bending movement. Tetrathiofulvlene-7,7,8,8-tetracyanoquinodimethane (TTF-TCNQ) and 2,2′-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS) electron transfer mediators are cast on the surface of the polypyrrole to help the electron transmission. The glucose oxidase and laccase enzymes are immobilized in the modified-conducting polymer surface, integrating the electrode to the actuator. The bio-catalysis of enzymes in presence of glucose and oxygen in aqueous solution provides the actuator with the electrons needed for the redox reaction, converting the chemical energy into mechanical energy, i.e., movement. The glucose-self-powered soft actuator may contribute to the development of more complex implantable, ingestible, or wearable biomedical devices such as cardio-stimulators, insulin pumps, or muscle implants.