The invention discloses a preparation method of modified anti-felting wool fiber. The method particularly comprises the following steps that 1, an oxidative antibacterial solution is prepared; 2, theoxidative antibacterial solution is added into a stirring tank, water bath heating is conducted to reach 40-45 DEG C, and the wool fiber is poured into the stirring tank; 3, compound enzymatic hydrolysate is prepared; 4, the wool fiber processed in step 2 is soaked in the compound enzymatic hydrolysate, and a normal temperature enzymatic hydrolysis reaction is conducted; 5, a chitosan solution isprepared and added into the stirring tank, the wool fiber prepared in step 4 is added, stirring is conducted for 5 h and then fished out, and then air-drying is conducted at normal temperature. Accordingly, the wool fiber is oxidized, protein of the wool fiber is exposed, the exposed protein is subjected to enzymolysis through protease, by means of enzymolysis, negative ions on the surface of thewool fiber are increased, chitosan is adsorbed to the surface of the wool fiber conveniently, and the texture is soft while the wool fiber anti-felting property is improved.