The invention discloses a construction method of an enzyme-reponsive multifunctional nano-coating. The construction method comprises the following steps: firstly, immobilizing avidin molecules to the surface of dopamine-coated stainless steel; then, on the basis of specific recognition and binding effects between avidin and biotin, immobilizing biotinylated heparin / PEI nanoparticles to the surface of a material, and further binding high-concentration avidin molecules to residual biotin on the surface of a nanoparticle coating, so that a novel biotin binding site is introduced; then, continuing to assemble biotinylated enzyme-responsive polypeptide to the surfaces of the nanoparticles on the basis of an interaction between the biotin and the avidin; and finally, by virtue of an EDC / NHS / MES coupling agent, immobilizing SDF-1[alpha] to an amino terminal of the polypeptide in a covalent mode, so that the multifunctional nano-coating, which has a matrix metalloproteinase 9 responsive characteristic, is constructed. According to the construction method provided by the invention, by constructing a multifunctional layer, which has anti-coagulating and endothelial regeneration inducing capacities, on a titanium surface, blood compatibility and a damaged endothelial repair capacity of the material can be obviously improved.