Novel biocatalysts that feature enzymes immobilized onto solid supports have recently become a major research focus in an effort to create more sustainable and greener chemistries in catalysis. Many of these novel biocatalyst systems feature enzymes immobilized onto metal-organic frameworks (MOFs), which have been shown to increase enzyme activity, stability, and recyclability in industrial processes. While the strategies used for immobilizing enzymes onto MOFs can vary, the conditions always require a buffer to maintain the functionality of the enzymes during immobilization. This report brings attention to critical buffer effects important to consider when developing enzyme/MOF biocatalysts, specifically for buffering systems containing phosphate ions. A comparative analysis of different enzyme/MOF biocatalysts featuring horseradish peroxidase and/or glucose oxidase immobilized onto the MOFs UiO-66, UiO-66-NH2, and UiO-67 using a noncoordinate buffering system (MOPSO buffer) and a phosphate buffering system (PBS) show that phosphate ions can have an inhibitory effect. Previous studies utilizing phosphate buffers for enzyme immobilization onto MOFs have shown Fourier transform infrared (FT-IR) spectra that have been assigned stretching frequencies associated with enzymes after immobilization. Analyses and characterizations using zeta potential measurements, scanning electron microscopy, Brunauer-Emmett-Teller surface area, powder X-ray diffraction, Energy Dispersive X-ray Spectroscopy, and FT-IR show concerning differences in enzyme loading and activity based on the buffering system used during immobilization.
© 2023 The Authors. Published by American Chemical Society.