Unravelling the Potential of Crude Enzyme Extracts for Biocatalyst Entrapment in Metal-Organic Frameworks

To bolster the applicability of enzymes as catalysts, it is imperative not only to address their inherent fragility, particularly when used under harsh organic-synthetic reaction conditions, but also to mitigate deactivation during purification and enable applicability in a broad range of organic-synthetic transformations. Currently, the process of purification of crude enzyme extracts and subsequent heterogenization to obtain immobilized biocatalysts often leads to partial enzyme deactivation and represents, at least in part, a resource-intensive process that is driving up the overall production efforts. To tackle both the enzyme fragility and deactivation during purification and immobilization, we propose the direct use of crude enzyme extracts obtained from cell lysis instead of pure enzymes and their entrapment in metal-organic framework (MOF) structures. We focus on three enzyme types with varying sensitivities: aldoxime dehydratase, imine reductase, and lipase. We evaluate the effects of different metal sources (Al, Fe, Co, Ni, Cu, and Zn), their oxidation state and counterions, and MOF synthesis parameters on enzyme stability and activity during their entrapment in the MOF structures. Based on this, we optimize protocols for enzyme entrapment in Fe-MIL-88A, Fe-MIL-100, Zn-MOF-74, and Zn-ZIF-8 and develop a fast-aqueous room temperature synthesis of Al-MIL-53. Investigation of the biocatalytic performance of the enzyme@MOF biocomposites suggests that enzyme entrapment in MOFs using crude enzyme extracts can effectively maintain enzyme activity and stability in various catalytic reactions, offering a perspective for an efficient pathway for industrial applications.