The rational construction of semiconductor nano-heterostructures is a feasible strategy to modulate electronic structure and increase active area of the electrocatalysts for biosensing. Herein, we develop an in situ approach, electrochemical (EC-) rebuilding of the smooth Cu surface, to construct hierarchical Co(OH)(2) nanosheets/CuO microcoral arrays (Co(OH)(2) NSs/CuO MCAs). Through engineering the heterostructures by optimizing EC-rebuilding time, the electrocatalytic activity is significantly enhanced with a higher current density of glucose oxidation. The incorporation of Co(OH)(2) NSs into CuO MCAs also leads to a large active surface area and benefits surface/interface reactions and mass transport for shorter response for glucose oxidation, higher current density, and better selectivity for glucose sensing. Both photoelectron spectra and density functional theory (DFT) calculations prove that interface charge transfers from CuO to Co(OH)(2), resulting in electron redistribution and a significant increase in the adsorption energy of glucose. Compared with recently reported enzyme-free glucose sensors, the fabricated Co(OH)(2) NSs/CuO MCAs electrode exhibits excellent performance for enzyme-free glucose-sensing in alkaline electrolytes with a short response time (3 s), wide linear range of 500 nM to 2.311 mM, ultrasensitivity of 2269 mA mM(-1) cm(-2), low limit of detection (LOD, 378 nM), and favorable reproducibility and stability. Noticeably, the outstanding response time, favorable ultrasensitivity, and great LOD are achieved in the glucose sensing. Therefore, the proposed sensor can be used for accurate quantification of glucose concentration in human serum with good repeatability, which will provide a new platform based low-cost semiconductor nano-heterostructures for rapid diagnostic tests and health monitoring.