Cascade‐Amplified Time‐Resolved Fluorescent Assay Driven by an Enzyme‐Integrated Catalytic Compartment as an Artificial Multi‐Enzyme Complex

We here report a simple and efficient strategy of fabricating artificial multi‐enzyme complex (MEC) based on the integration of natural enzyme with catalytic compartment. As a proof of concept, this strategy was demonstrated by selecting cholesterol oxidase (ChOx) and CeIII‐based nanoscale coordination polymer (Ce‐NCP) with peroxidase‐like activity as the models, which forms ChOx@Ce‐NCP. Benefitting from the confinement and sheltering effects of Ce‐NCP, superior cascade activity and stability in harsh environments were achieved in ChOx@Ce‐NCP. Meanwhile, the distinct advantage of ChOx@Ce‐NCP has also been highlighted by its negligible substrate inhibition effect and adjustable mass ratio of building blocks. Upon the doping of TbIII in ChOx@Ce‐NCP, a luminescent artificial MEC (ChOx@Ce‐NCP:Tb) was further fabricated to drive a cascade amplified time‐resolved fluorescent assay within a confined space, showing high sensitivity and specificity toward cholesterol.