Facile construction of multicompartment multienzyme system through layer-by-layer self-assembly and biomimetic mineralization.

In nature, some organelles such as mitochondria and chloroplasts possess multicompartment structure, which render powerful and versatile performance in cascade conversion, selective separation, and energy transfer. In this study, mitochondria-inspired hybrid double membrane microcapsules (HDMMCs) were prepared through synergy between biomimetic mineralization and layer-by-layer (LbL) self-assembly using double templating strategy. The organic inner membrane was acquired via LbL self-assembly of oxidized alginate (o-alginate) and protamine on the CaCO(3) template, the silica template layer was then formed onto the inner membrane through biomimetic silicification using protamine as inducer and silicate as precursor, the organic-inorganic hybrid outer membrane was acquired via biomimetic mineralization of titanium precursor. After the CaCO(3) template and the silica template are removed subsequently, multicompartment microcapsules with microscale lumen and nanoscale intermembrane space were obtained. The double membrane structure of the HDMMCs was verified by high resolution scanning electron microscopy (HRSEM), and the superior mechanical stability of HDMMCs was demonstrated by osmotic pressure experiment and fluorescence microscopy. A multienzyme system was constructed by following this protocol: the first enzyme was encapsulated in the lumen of the HDMMCs, whereas the second enzyme was encapsulated in the intermembrane space. Compared to encapsulated multienzyme in single-compartment microcapsules (SCMCs) or in free form in aqueous solution, enzymatic activity, selectivity, and recycling stability of HDMMCs-enabled multienzyme system were significantly improved. Because of the inherent gentle and generic feature, the present study can be utilized to create a variety of compartment structures for the potential applications in chemical/biological catalysis and separation, drug/gene delivery systems, and biosensors.