Both physiological behaviors and pathological processes of living organisms are related to the synergistic co-action of various enzymes. Inspired by this, a new bionic catalytic technology, immobilized multi-enzyme cascade reactor has been proposed for accelerating the bio-reactions including synthesis, substrate degradation, transformation and recombination. By taking advantages of the good stability, reusability and remarkable high efficiency, the immobilized multi-enzyme cascade reactor has attracted more and more attention in many fields including biological sensing, simulation biology and biological transformation and so on. This review focuses on the art-of-state and progress of immobilized multi-enzyme cascade reactor in recent years. The fundamentals, preparation approaches, advantages, factors affecting the efficiency, and applications of the technology are involved. Its trends in future study are also prospected.