Au/TiO2 composites were synthesized by a two-step hydrothermal approach and their photocatalytic and electrochemical properties were investigated. It is found that Au/TiO2 composites with a large specific surface area, stability and catalytic activity of Au nanoparticles (NPs) demonstrate enhanced photocatalytic activity compared to pure TiO2 microspheres. Furthermore, the electrochemical properties of the Au/TiO2 composite electrode demonstrate its ability to electrocatalyze the oxidation of hydrogen peroxide and substantially increase the response current. In addition, this modified electrode also exhibits a very rapid and sensitive response towards glucose detection at the low operating potential. The modified electrode shows a linear response from 1.0 to 15 mM with a reproducible sensitivity of 1350 μA mM−1 cm−2 within a response time of less than 6 s. The photocatalytic and electrochemical activity of Au/TiO2 composites provides a new platform for environmental remediation and biomedical applications. All these results reveal that Au/TiO2 composites exhibit great prospects for developing efficient non-enzymatic biosensors.