Mechanistic Understanding of Protein–MOF Integration Through Surfactant‐Driven Interfacial Design

ABSTRACT Integration of proteins into metal–organic frameworks (Protein@MOF) offers an effective strategy for protein stabilization across materials and biomedical applications. However, the molecular mechanism of protein–MOF interactions remains poorly understood, limiting rational control over their chemical and physical properties. Here, we develop a surfactant‐guided strategy to modulate the assembly of protein@MOF through interfacial design. We discovered that the interfacial environment between proteins and MOFs is the dominant factor controlling encapsulation efficiency, structural integrity, and functional performance. Lipid‐based non‐ionic surfactants such as glycerol monooleate (GMO) increase the protein's solvent‐accessible surface area (SASA), suggesting partial remodeling of the protein surface and hydration shell. Interfacial GMO enhances protein encapsulation by 20% and accelerates MOF growth by 30%. Importantly, for horseradish peroxidase (HRP) encapsulated in MOF, incorporation of lecithin results in up to a six‐fold enhancement in retained bioactivity and a near 60‐fold increase in k _cat . All‐atom molecular dynamics simulations reveal concentration‐dependent, domain‐specific interactions between the surfactant and flexible surface residues via electrostatic and hydrophobic contacts. These findings establish surfactant‐driven interfacial design as a general molecular strategy to enhance protein@MOF stability and function, enabling robust alternatives to lipid nanodiscs for membrane protein stabilization and advancing applications in biocatalysis, biosensing, and drug delivery.