Abstract: In order to obtain a new type of bacterial cellulose-based composite with antibacterial activity, by using the ultrasonic vibration, the PLBC composite membrane was prepared by making the bacterial cellulose (BC) obtained by fermentation fully contact adsorption with ε-Polylysine (ε-PL). The hygroscopic rate, dehydration rate, microstructure and mechanical properties of PLBC composite membrane were investigated, and the performance characterization of PLBC composite membrane was analyzed by using the infrared spectroscopy and XRD analysis. Then, the antibacterial activity of PLBC composite membrane on E.coli and S.aureus was investigated by using the bacteriostatic circle method and absorbance method, and the antibacterial mechanism of PLBC composite membrane on E.coli and S.aureus was preliminarily discussed. The results showed that the crystallization of ε-PL would affect the microstructure of PLBC composite membrane, and the appropriate concentration of ε-PL was 0.5% in the preparation of PLBC composite membrane. The results of the infrared spectroscopy and XRD analysis proved that the PLBC composite membrane could be successfully prepared. The mechanical properties of PLBC composite membrane have been improved to a certain extent compared with BC membrane. At the same time, because of the introduction of ε-PL, PLBC composite membrane had good antibacterial activity, which was a kind of medical dressing with application prospects or antibacterial food packaging material.