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A series of Cu-MCM-41 catalysts were synthesized through in-situ hydrothermal preparation method, calcinated at different temperatures and characterized by thermogravimetric and differential scanning calorimetry (TG-DSC), N2 adsorption-desorption, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FI-IR), transmission electron microscopy (TEM) and hydrogen temperature-programmed (H2-TPR). The effect of calcination temperature on the structural properties and selective oxidation of dimethoxymethane (DMM) to dimethyl carbonate (DMC) were discussed in detail. The results showed that the calcination temperature played important role on the microstructure and catalytic activity of the Cu-MCM-41 catalysts. At the calcination temperature 550 °C, the Cu-MCM-41 catalyst possessed higher surface area, smaller pore diameter, stronger metal-support interaction, and better CuO dispersion compared the other catalysts, and the corresponding catalyst exhibited excellent activity and stability for the DMC synthesis. Under the reaction conditions at 2.0 MPa and 130 °C, the highest DMM conversion was 99.47% with the best DMC selectivity of 85.01%. In addition, the catalyst was reused four times without significant loss of performance.