DNA-directed assembly of core-satellite MOF structures for spatially compartmentalized multi-enzyme cascade catalysis

The spatial organization of enzymes in natural cells enables efficient metabolic cascades through localized microenvironments and controlled intermediate diffusion. Mimicking this compartmentalized architecture in artificial systems remains a key challenge in biocatalysis. Here, we introduce a DNA-mediated self-assembly strategy to construct the biomimetic core-satellite structure of metal-organic frameworks (MOFs) that spatially organize multiple enzymes into defined compartments. In this system, DNA bridges larger “core” MOFs and smaller “satellite” MOFs, forming ordered assemblies with tunable microenvironments tailored for individual enzymes. Using glucose oxidase (GOx) and horseradish peroxidase (HRP) as a model cascade, we demonstrate that the core-satellite structure facilitates substrate diffusion, mitigates inhibition by intermediates, and enhances overall catalytic efficiency. This platform was further applied to support multi-enzyme cascades, including the two-enzyme system of N -acetyl hexosamine 1-kinase (NahK)/UDP- N -acetylgalactosamine pyrophosphorylase (GlmU) for glycosyl donor synthesis, and the three-enzyme system of β-galactosidase (β-Gal)/GOx/HRP for sequential cascade catalysis. Compared to the single MOF system, spatially compartmentalized MOFs exhibit up to 34-fold higher activity and improved thermal, solvent, and operational stability. This work provides a versatile platform for the design of cell-inspired biocatalytic systems and underscores the potential of DNA-guided MOF assembly in constructing programmable multi-compartment enzyme reactors.