Electrospinning provides an effective method for generating nanofibers from solution of carboxymethyl chitosan/polyoxyethylene oxide (CMCS/PEO). The goal of this work is to explore the potential application of electrospun CMCS/PEO nanofiber membrane in fruit fresh-keeping. The microstructure, antibacterial activity, hydrophilia, and air permeability of the nanofiber membrane have been tested. For comparison, the fresh-keeping effects of commercial cling wrap and CMCS/PEO nanofiber membranes on strawberries' rotting rate and weight loss rate have been studied. The results indicate that the electrospun CMCS/PEO membrane could effectively avoid water loss in strawberries and has a remarkable effect to prolong strawberries' shelf life due to its breathability and antibacterial activity. In addition, the composite CMCS/PEO, nanofiber membrane is non-poisonous and edible, which can be used in food industry.