Heterostructure Cu₂O@CuO nanosheets are spontaneously formed during the kinetic-spraying of nanostructured CuO thin film on the fluorine-doped tin oxide (FTO) substrate at room temperature under low vacuum conditions by the NPDS technique. The Cu₂O@CuO nanosheets fabricated as a heterogeneous electrocatalyst are utilized for free enzyme detection of glucose and H₂O₂ in 0.1 M NaOH, whereby the synergy between the multivalent copper species (I and II) results in an overall improvement of electrocatalytic performance. The in-situ formation of interfacial Cu₂O@CuO heterostructure is verified using a high-resolution transmission microscope and x-ray photon electron spectroscopy. The fabricated electrocatalysts exhibit high sensitivity toward glucose oxidation of 818.5 μA·mM^-1·cm^-2 with a wide linear detection range extending from 0.02 to 5 mM and a limit of detection (LOD) of 5.8 μM. Also, the heterostructure Cu₂O@CuO nanosheets show a very wide linear detection range of H₂O₂ oxidation from 0.02 to 11 mM with a good detection sensitivity of 168 μA·mM^-1·cm^-2 and a LOD of 8.75 μM. The long-term stability for around 30 min and high oxidation selectivity with other interfering species are verified. Furthermore, Cu₂O@CuO nanosheets exhibit higher catalytic rate constant and diffusion coefficient toward glucose species compared with H₂O₂.