【Purposes】 Combining the high selectivity of natural enzymes with the high stability of nanoenzymes in a one-pot reaction has emerged as an ideal solution to improving the diversity/complexity of biocatalytic cascades and the stability of multi-enzyme systems. 【Methods】 In this paper, we designed and synthesized an ionic magnetic Fe3O4@EB-COFs material to prepare glucose oxidase (GOx)-magnetic covalent organic framework material(GOx/Fe3O4@EB-COFs) through electrostatic interaction induction. 【Findings】 Fe3O4@EB-COFs played the role of nanoparticle enzyme and carrier material, and the obtained composite could be used for glucose concentration for colorimetric determination. Under acidic conditions, GOx/Fe3O4@EB-COFs exhibited a good cascade of catalytic glucose oxidation and 2,2′-biazo-bis-3-ethylbenzothiazoline-6-sulfonic acid oxidation. The kinetic study showed that the reaction followed the Lineweaver-Burk equation, indicating that the cascade reaction of the constructed nanoenzyme with natural enzyme still exhibited biocatalytic characteristics. GOx/Fe3O4@EB-COFs exhibited good recoverability with the addition of magnetic Fe3O4. 【Conclusions】 Therefore, the construction of a multi-enzyme catalytic system of nanoenzymes and natural enzymes is important for the realization of biomimetic catalysis.