A highly sensitive glucose biosensor was fabricated by immobilizing the glucose oxidase(GOD) on a new hybrid film of Au nanoparticles(Au NPs) and hydroxyapatite(HAp).The direct electrochemical behavior of GOD in the hybrid film was enhanced and a pair of well-defined，nearly reversible peaks appeared in deoxygenized pH 7.0 PBS due to the presence of AuNPs.The experimental results also showed that the peak current of dissolved oxygen at -0.8 V decreased with the addition of glucose due to the consumption of dissolved oxygen when the glucose was oxidized by GOD.The decreased reduction current of oxygen was linear with concentration of glucose in the range of 0.02-1.62 mmol/L with a sensitivity of 9.91 mA?mol-1?L and a detection limit of 5.0 μmol/L.The biosensor could be applied in the determination of glucose with high sensitivity and speediness.