On the Assembly of Microreactors with Parallel Enzymatic Pathways

Cell mimicry aims to create artificial structures capable of mimicking certain functions of living cells. However, cell mimics are tremendously simpler than their natural role models. Thus, increasing their complexity is of great importance for the advancement of cell mimicry concepts and for further understanding of biological intracellular processes. Here, the successful co-encapsulation of two enzymatic pathways with up to five enzymes into compartmentalized microreactors is reported. The microreactors are assembled by combining polymer layers and enzyme-loaded liposomal subunits, which physically separate the two encapsulated enzymatic pathways. Specifically, this report confirms the activity of an encapsulated enzymatic cycle that conjugates the actions of glutamate dehydrogenase and glutathione reductase, using NADP⁺ /NADPH as a common co-factor, as well as an encapsulated enzymatic cascade combining β-galactosidase, glucose oxidase, and catalase. This work represents a relevant advancement in encapsulated catalysis toward the assembly of therapeutic cell mimics.