Fabricating compartmented nanoreactor via protein self-regulated biomimetic mineralization for multienzyme immobilization

Constructing compartmented nanoreactors for the sequential immobilization of multienzyme cascade systems still presents formidable challenges. Here, inspired by biomolecule-mediated biomimetic mineralization, a compartmented nanoreactor (named CoMIm@CaP) was constructed by protein self-regulated biomimetic mineralization. Multienzyme cascade systems consisting of sucrose phosphorylase (SPase) and D-allulose 3-epimerase (DAE) could be compartmentally immobilized in CoMIm@CaP to form DAE/CoMIm@SPase/CaP with robust stability and superior activity. Remarkably, the catalytic activity of DAE/CoMIm via protein self-regulated biomimetic mineralization was 12.5-fold than that of DAE/ZIF-67 passively encapsulated in common ZIF-67. About 123 mg/mL of 2-O-α-GG and 38 mg/mL of D-allulose were produced from high concentrations of sucrose and glycerol via DAE/CoMIm@SPase/CaP catalysis. Our findings introduce a versatile means to construct a compartmented nanoreactor for the partitioning immobilization of multienzyme.