Flexible magnetic membrane based on bacterial cellulose (BC) was successfully prepared by in-situ synthesis of the Fe3O4 nanoparticles under different conditions and its properties were characterized. The results demonstrated that the Fe3O4 nanoparticles coated with PEG were well homogeneously dispersed in the BC matrix under ultrasonic irradiation with the saturation magnetization of 40.58 emu/g. Besides that, the membranes exhibited the striking flexibility and mechanical properties. This study provided a green and facile method to inhibit magnetic nanoparticle aggregation without compromising the mechanical properties of the nanocomposites. Magnetically responsive BC membrane would have potential applications in electronic actuators, information storage, electromagnetic shielding coating and anti-counterfeit.