Nano-zinc oxide is a kind of inorganic antibacterial agent, which had good antibacterial effect and didn’t produce drug resistance. Clove oil is a kind of natural antibacterial agent with spectral antibacterial properties and non-toxic health. In this paper, chitosan and konjac glucomannan were used as the film-forming substrates, two kinds of antibacterial agent were added to the biomass composite membrane. The antibacterial rate of the composite membrane against Escherichia coli and Staphylococcus aureus was 99.78% and 99.06%, respectively. The film had a strong antibacterial effect. The results of infrared spectroscopy indicated that the chitosan functional group and the phenol group in the clove oil were effectively bonded, and the results of scanning electron microscopy showed that the clove oil improved the dispersion of nano zinc oxide. The composite membrane was used for the preservation experiment of fresh meat. The preservation effect of the composite membrane was evaluated in a 14-day cycle. It was found that the composite membrane with nano zinc oxide and clove oil increased the shelf life of fresh meat by 6~8 days.