AbstractSome cellular enzymatic pathways are located within a single organelle, while most others involve enzymes that are located within multiple compartmentalized cellular organelles to realize the efficient multi‐step enzymatic process. Herein, bioinspired by enzyme‐mediated biosynthesis and biochemical defense, a compartmented nanoreactor (Burr‐NCs@GlSOD) was constructed through a self‐confined catalysis strategy with burr defect‐engineered molybdenum disulfide/Prussian blue analogues (MoS2/PBA) and an interfacial diffusion‐controlled hydrogel network. The specific catalytic mechanism of the laccase‐like superactivity induced hydrogelation and cascade enzyme catalytic therapy were explored. The confined hydrogelation strategy introduces a versatile means for nanointerface functionalization and provides insight into biological construction of simulated enzymes with comparable activity and also the specificity to natural enzymes.