Organic Molecular Cage-Mediated pH-Responsive Reversible Protein Self-Assembly with Multi-Enhanced Enzymatic Performance

Despite the various advantages and versatile applications of protein self-assemblies, their use as immobilized enzymes in biocatalysis, especially for organic synthesis, has rarely been reported. Here, we achieved the pH-responsive reversible self-assembly of an industrially important amine dehydrogenase (AmDH) with the assistance of porous organic molecular cages (OMCs). The thus-formed AmDH–OMC self-assemblies as heterogeneous biocatalysts demonstrated the following advantages that are highly challenging to conventional immobilization strategies: (1) multiperformance enhancements, including stability, activity, kinetics, and catalytic efficiency; (2) high recoverability and recyclability of both the enzyme and the OMCs; and (3) mechanistic elucidation of the AmDH–OMC intermolecular interaction at the molecular level by molecular dynamics simulations. This strategy also enabled the in situ preparation and application of multienzyme and enzyme–metal coimmobilized biocatalysts for the enantioselective synthesis of chiral amines in batch and flow systems, demonstrating its high synthetic utility.