Biofilms adhering to device surfaces impose great threat to public health and industries. Metal-organic frameworks (MOFs) replicating the functionalities of natural enzymes are highly promising for combating biofilms, however still challenged by the intrinsic inferior catalytic activity. Herein, a simple and facile strategy is demonstrated to fabricate ultrasmall Ce-MOFs in an aqueous solution at ambient temperature to achieve high hydrolytic activity under mild conditions. Benefiting from the biomimetic structure and the ultrasmall size, Ce-MOFs nanozyme exhibited remarkable hydrolytic catalytic activities for cleaving the phosphate bonds, glycosidic bonds and their mixtures, the biofilm, showing satisfactory performance in biofilm prevention and eradication. Catalytic mechanism investigation reveals that the synergistic interaction between Ce4+ and Ce3+, the dense active site leads to 3–15 folds higher hydrolytic activity of the ultrasmall sized Ce-MOFs compared with the bulk ones. This work is expected to provide insights for rational design of nanozymes and holds great promise for environmental remediation.