One-pot spatial engineering of multi-enzymes in metal–organic frameworks for enhanced cascade activity

Abstract

Enzyme encapsulation in metal–organic frameworks (MOFs) faces significant challenges, including restricted substrate diffusion, random enzyme distribution, and diminished catalytic performance, particularly in multi-enzyme systems. To address these issues, we propose a facile one-pot strategy to achieve spatial engineering of multiple enzymes in MOFs. Using the environmentally sustainable and cost-effective polymer, poly(diallyldimethylammonium chloride) (PDADMAC), the surface charge of enzymes can be selectively modified. This modification allows enzymes with a negative surface charge to accelerate MOF nucleation, leading to their primary localization in the core, while enzymes with a positive surface charge slow the nucleation process, resulting in their exclusive placement in the outer shell. This structure enables effective compartmentalization, efficient intermediate transfer, prevention of cross-reactivity, and alteration of enzyme conformation. Consequently, this approach significantly enhances the cascade reaction activity and is effective even for incompatible enzymes, achieving a 1.69- to 14.85-fold improvement in catalytic efficiency compared to unmodified systems. Additionally, the resulting biocomposites exhibit robust catalytic performance under adverse environmental conditions and maintain their efficiency over multiple cycles, making them highly promising for diverse industrial and environmental applications.