To rapidly detect glucose, the immobilized dual-enzymes cascade reactor was designed by co-immobilizing glucose oxidase (GOx) and horseradish peroxidase (HRP). The micromorphology and chemical composition of sodium alginate hydrogel were analyzed by using the scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Meanwhile, the immobilized dual-enzymes co-catalytic system was optimized and applied for the detection of glucose in the beverage sample. Based on the enzyme-catalyzed reaction, an efficient and specific glucose detection method was established by using the cascade reaction of co-immobilized enzymes, which provids a promising strategy to design bio-detechion experiments.