Aspartic Acid-Mediated Potential Matching Directs Assembly of MOF-Enzyme Composites Featuring Accessible Active Sites for Enhanced Biocatalysis

The random adsorption of enzymes onto immobilized carrier interfaces often leads to conformational distortion of catalytically active sites, structural mismatch of substrate-binding pockets, and blockage of mass transfer channels, thereby severely constraining catalytic efficiency. Herein, this study proposes a computationally guided interfacial engineering strategy based on aspartic acid-mediated potential matching. By introducing aspartic acid residues at key interface positions to regulate interface potential, proteins can spontaneously adsorb onto metal–organic framework (MOF) carriers’ surface in optimal conformation, achieving high controllability of the spatial orientation of enzyme molecules (>90% active site orientation probability). Consequently, this strategy increased the catalytic activity of β-glucosidase (β-G) by 4.21-fold and the microscopic mass-transfer efficiency by 5.64-fold compared with the nonmodified system and ultimately achieved 86.6% of the total conversion efficiency of ginsenoside compound K (CK). This work demonstrates the role of interfacial potential matching in simultaneously stabilizing enzyme conformation and maximizing mass-transfer accessibility, providing a universal platform for precision enzyme immobilization.