The work aims to develop a novel biological preservation film with excellent physical and chemical properties and good preservation effect. Pullulan and konjac glucomannan (KGM) were used as the base materials and then added with different concentrations of fish collagen to prepare novel composite biological preservation film. The effects of fish collagen on the physical and biological properties of the composite film were analyzed by measuring water solubility, light transmittance and texture, and the preservation effect of the composite film on grass carp during low temperature storage was discussed by taking the values of TVB-N, pH and total bacteria on the surface of fish fillet as indexes. Fish collagen could improve the physical and chemical properties of polysaccharide preservation film and the preservation effect was the best when its mass fraction was 3%. After the grass carps treated with different films were stored at 4 ℃ for 12 d, the film with 3% fish collagen could delay the spoilage of fish meat, the breeding of food-borne pathogens and the generation of volatile base nitrogen in meat products and effectively inhibited the oxidative deterioration of lipids and protein. The use of 3% fish collagen can improve the physical and biological properties of polysaccharide-based biological preservation film, significantly maintain the texture properties, and extend the shelf life of grass carp from 4 d to 9~10 d. This novel composite preservation film has good application value and development prospects.