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• W3023285493 startingPage "22640" @default.
• W3023285493 abstract "Polyelectrolyte multilayers (PEMs) are attractive materials for immobilizing enzymes due to their unique ionic environment, which can prevent unfolding. Here, we demonstrated that the stability to dry storage and elevated pH were significantly enhanced when negatively charged nitroreductase (NfsB) was embedded in a PEM by depositing alternating layers of the enzyme and polycation (PC) onto porous silica particles. The PC strength (i.e., pKa) and the surface charge of the film were varied to probe the effects that internal and surface chemistry had on the pH stability of the entrapped NfsB. All films showed enhanced activity retention at elevated pH (>6), and inactivation at reduced pH (<6) similar to NfsB in solution, indicating that the primary stabilizing effect of immobilization was achieved through ionic interactions between NfsB and the PC and not through changes to the surface charge of the NfsB. Additionally, films that were stored dry at 4 °C for 1 month retained full activity, while those stored at room temperature lost 30% activity. Remarkably, at 50 °C, above the NfsB melting temperature, 40% activity was retained after 1 month of dry storage. Our results suggest that internal film properties are significantly more important than surface charge, which had minor effects on activity. Specifically, immobilization with the weak PC, poly(l-lysine), increased the optimal pH and the activity of immobilized NfsB (which we attribute to greater permeability), relative to immobilization with the strong PC, poly(diallyldimethylammonium chloride). However, NfsB was leached from the PLL film to a greater extent. Overall, these observations demonstrate that internal ionic cross-linking is key to the stabilizing effects of PEMs and that the pH response can be tuned by controlling the number of cross-links (e.g., by changing the strength of the PC). However, this may be at the cost of reduced loading, illustrating the necessity of simultaneously optimizing enzyme loading, internal ionic cross-linking, and substrate transport." @default.
• W3023285493 created "2020-05-13" @default.
• W3023285493 creator A5051824598 @default.
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• W3023285493 date "2020-04-30" @default.
• W3023285493 modified "2023-09-24" @default.
• W3023285493 title "Polyelectrolyte Multilayers Enhance the Dry Storage and pH Stability of Physically Entrapped Enzymes" @default.
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• W3023285493 doi "https://doi.org/10.1021/acsami.0c04964" @default.
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