The controlled spatial organization or compartmentalization of multi-enzyme cascade reactions to transfer a substrate from one enzyme to another for substrate channeling on scaffolds has sparked increasing interest in recent years. Here, we use graphene oxides to study the dependence of the activity of cascade reactions in a closely packed, randomly immobilized enzyme system on a 2 D scaffold. We first observe that the hydrophobicity of graphene oxides and various enzyme architectures for co-immobilized systems are important attributes for achieving high product-conversion rates. A transient time close to 0 s can be achieved if enzymes are randomly immobilized close to one another, owing to direct molecular channeling. This contributes to overcoming complications regarding control of the spatial arrangement of the enzymes. Furthermore, a fabricated bienzyme paper can be used for glucose detection with high stability, reusability, and enhanced substrate channeling. Our findings provide new guidance for enzyme orientation on 2 D scaffolds, which may be extrapolated to other multienzyme cascade systems.
Keywords: enzyme immobilization; enzyme nanoarchitectonics; enzymes; graphene; substrate channeling.
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