As a kind of natural polymer material with good barrier and film-forming properties, cellulose has attracted wide attention in the application of packaging preservation film. In view of its limited application due to the lack of antibacterial properties, a simple two-step method has thus been proposed for the preparation of antibacterial cellulose membrane so as to obtain antibacterial properties quickly. The regenerated cellulose membrane is prepared by the sol-gel method, to be oxidized by sodium periodate to obtain the oxidized regenerated cellulose membrane (DRCF). Meanwhile, an investigation is made of the influence of oxidation on regenerated cellulose membrane, with their antibacterial properties tested and characterized. The experimental results show that the oxidation of sodium periodate successfully generates aldehyde groups in the regenerated cellulose membrane, while the aldehyde group content and oxidation degree gradually increase positively with the sodium periodate concentration and oxidation time. Sodium periodate oxidation increases the tensile strength of the membrane, reaching 102.9 MPa after 6 hours of oxidation. The growth of Staphylococcus aureus can be inhibited by DRCF within 24 h, thus achieving the effect of killing bacteria, with an additional inhibitory effect on the growth of Escherichia coli as well. It can be seen that the cellulose membrane prepared by the proposed method, characterized with good antibacterial properties, meets the mechanical performance requirements for packaging membrane use, which can be applied in the field of antibacterial food packaging.