Abstract: Objective:To replace the traditional non-renewable plastic wrap, the performance of antibacterial plastic film was studied using the economical and environment-friendly raw materials. Methods:Under the best film-forming conditions, the effects of the concentrations of pullulan polysaccharides, sophorolipids and glycerin on the properties of preservative film were analyzed by single factor experiments. Meantime, the optimal inhibitory concentration of sophorolipids was investigated. Then, their suitable concentrations were determined by orthogonal experiment. After that, the fresh-keeping property of preservative solution and film were investigated. Results:The antimicrobial activity of sophorolipids on Staphylococcus aureus was significant (p<0.01). It was 100% inhibited when the concentration of sophorolipids was 2.4 g/L.The sophorolipids have different inhibition on Escherichia coli and Candida albicans. When the concentration of sophorolipids was 4.0 g/L, the inhibition rate against E. coli and C. albicans were 28.75% and 14.53%, respectively. The best formula of preservative solution was determined as 18 g/L of pullulan polysaccharides, 3.6 g/L of sophorolipids and 8.75 g/L of glycerin. Both the preservative solution and the preservative film have good effects on fresh keeping of cherries (p<0.01).Conclusion:Using bio-surfactant sophorolipids as raw materials of antimicrobial preservative film could provide a new theoretical basis for the preparation process of green antimicrobial cling film.