Abstract: N, O-carboxymethyl chitosan (N, O-CMC) with high substitution was prepared at 80℃ by mixing chitosan and monochloroacetic acid with different proportions under alkaline condition. Infrared Spectrum (FTIR) was used to identify the structure of N, O-CMC, and Potentiometric Titration was used to measure the substitution degree of N, O-CMC. N, O-CMC. Bamboo viscose cellulose and N, O-CMC was mixed by different proportions in the solution system of NaOHureathioureawater to get bamboo fiber film. The antibacterial property of bamboo fiber film against Escherichia coli and Staphylococcus aureus was measured by textiles evaluation for antibacterial activityShake Flask Method (GB/T 20944.3-2008). The results showed that the N, O-CMC added bamboo fiber film had good antibacterial property against Escherichia coli and Staphylococcus aureus. The antibacterial property of the blend membrane was improved when the proportion of N, O-CMC was increased.