A novel electrochemical sensor based on molecularly imprinted polymer membranes (MIPM) as biomimetic molecular recognition element involved in o-aminothiophenol functionalized Au nanoparticles (ATP@AuNPs) modified gold electrode was constructed for sensitive and selective detection of herbicide simazine (SMZ). The nano-scaled MIPM, with high specific surface area, was prepared by self assembly of o-aminothiophenol (ATP) and electrodeposition of ATP@AuNPs in the presence of template SMZ. Cathodic current of SMZ was measured by cyclic voltammetry and the results exhibited that the proposed sensor possess a high electrocatalytic activity at a negative potential and a fast rebinding dynamics towards the reduction of SMZ in 0.01 M H2SO4 solution (pH 1.7). Linear dependency of peak current on SMZ concentrations was observed from 0.03 to 140 M and detection limit was estimated to be 0.013 (3S/N). The enhancement of sensitivity was attributed to the presence of gold nanoparticles (AuNPs) which decreased the electron-transfer impedance and increased imprinting sites, lead to the superior enrichment effect of the trace amount of SMZ in aqueous solvent. The developed SMZ imprinted sensor exhibited excellent long-term stability and acceptable repeatability. In addition, the proposed method was successfully applied to measure SMZ in several real samples with the spiked recoveries changing from 91.4% to 96.8%, showing a promising potential in practical application.