TEMPO-oxidized bacterial cellulose (TOBC) that can be dissolved in water was fabricated through the treated bacterial cellulose using TEMPO (2, 2, 6, 6-tetramethylpiperidine-1-oxyl radical). Based on a cross-linked networking membrane of TOBC and polyvinyl alcohol (PVA), an electro active biopolymer actuator has been newly developed as artificial muscle. The conductive polymer PEDOT: PSS was used as the electrode of composite membrane using dipping and drying method. The obtained TOBC-PVA nanocomposite actuator exhibited larger bending deformation and faster time than those of TOBC actuator under both step and harmonic driving voltage. Properties of TOBC-PVA membrane were analyzed by employing such as scanning electron microscope (SEM), X-ray diffraction (XRD), water uptake, thermogravimetric analysis (TGA) and FTIR spectra. Accordingly, the TOBC-PVA nanocomposite membrane has better water uptake and thermal stability but more amorphous region, lower crystallinity and stiffness. Furthermore, the interfacial interaction between TOBC and PVA by cross-linking was showed by FT-IR spectra. The electro-mechanical performances of TOBC-PVA actuator exhibited much larger bending deformation owing to cross-linking and amorphous region and lower stiffness. Present results indicate that nanocomposite TOBC-PVA actuators with bio-compatibility and compatibility can be used for actuators, sensors biomimetic robotics and biomedical devices.