Enzymes are a class of natural catalysts with high efficiency, specificity, and selectivity unmatched by their synthetic counterparts and dictate a myriad of reactions that constitute various cascades in living cells. The development of suitable supports is significant for the immobilization of structurally flexible enzymes, enabling biomimetic transformation in the extracellular environment. Accordingly, porous organic frameworks, including metal organic frameworks (MOFs), covalent organic frameworks (COFs) and hydrogen-bonded organic frameworks (HOFs), have emerged as ideal supports for the immobilization of enzymes because of their structural features including ultrahigh surface area, tailorable porosity, and versatile framework compositions. Specially, organic framework-encased enzymes have shown significant enhancement in stability and reusability, and their tailorable pore opening provides a gatekeeper-like effect for guest sieving, which is beneficial for mimicking intracellular biocatalysis processes. This immobilization technique brings new insight into the development of next-generation enzyme materials and shows huge potential in healthcare applications, such as biomarker diagnosis, biostorage, and cancer and antibacterial therapies. In this review, we describe the state-of-the-art strategies for the structural immobilization of enzymes using the well-explored MOFs and burgeoning COFs and HOFs as scaffolds, with special emphasis on how these porous framework-confined technologies can provide a favorable microenvironment for mimicking natural biocatalysis. Subsequently, advanced characterization techniques for enzyme conformation, the effect of the confined microenvironment on the activity of enzymes, and the emerging healthcare applications will be surveyed.