The invention discloses a lactobacillus rhamnosus exopolysaccharide, and a preparation method and application thereof, and belongs to the technical field of microorganisms. According to the invention, a lactobacillus rhamnosus strain separated from Xiangshuangpao yoghurt is used as an original strain to produce exopolysaccharide, the exopolysaccharide is a uniform component, and the molecular weight of the exopolysaccharide is 88650 Da; and the lactobacillus rhamnosus exopolysaccharide comprises mannose, glucuronic acid, glucose, xylose, galactose and arabinose. The exopolysaccharide has a good in-vitro hypoglycemic effect, has good in-vitro alpha-glucosidase inhibition capacity through detection, and is simple in process, wide in market prospect, stable in thermodynamic property and suitable for large-scale production. Therefore, the polysaccharide is a potential active ingredient for preventing and treating diabetes mellitus. Besides, as a fermentation product of probiotics, compared with common chemical medicines in the field of blood sugar reduction, the fermentation product has the advantages of simple and easily available active ingredients, low price, no side effect and the like, and has great application value.