Trehalose hydrogels for stabilization of enzymes to heat

Abstract

Enzymes can catalyze various reactions with high selectivity and are involved in many important biological processes. However, the general instability of enzymes against high temperature often limits their application. To address this, we synthesized a trehalose-based hydrogel in two steps from commercial starting materials with minimal purification procedures. Mono- and multi-functional trehalose monomers were cross-linked by redox-initiated radical polymerization to form a hydrogel. Phytase, an important enzyme utilized in animal feedstock, was employed to study the effectiveness of the trehalose hydrogel to stabilize proteins against heat. Addition of the phytase solution to the hydrogel resulted in enzyme internalization as confirmed by confocal microscopy. The phytase in the hydrogel retained 100% activity upon heating at 90 °C compared to 39% when the hydrogel was absent. The enzyme could also be recovered from the hydrogel. The trehalose hydrogel synthesis reported herein should be readily scalable for thermal stabilization of a wide variety of enzymes.

Trehalose Glycopolymer Enhances Both Solution Stability and Pharmacokinetics of a Therapeutic Protein

“…The polymer has the disaccharide trehalose at the side chains and stabilizes various enzymes and proteins to fluctuations in temperature and lyophilization in solution. 21–23 We have also employed the trehalose glycopolymer as a resist material and demonstrated that polymer stabilizes proteins to high vacuum and direct electron beam irradiation in solid films. 24, 25 Trehalose is known to stabilize proteins with a mechanism attributed to vitrification, water replacement, and/or water entrapment.…”

Growth Factor-Loaded Microparticles for Tissue Engineering: The Discrepancies of In Vitro Characterization Assays

“…34 The ability of trehalose to stabilize biological molecules is generally highly valued; in fact, some animals release significant levels of trehalose in response to environmental stresses, as a protection mechanism, and hence many pharmaceutical drugs use trehalose as a stabilizer. 36 Trehalose has shown great advantages as an excipient in linear polymers for proteins 37 and enzymes 36 or as a conjugate to stabilize proteins and retain protein activity against high heat conditions and lyophilization. 38 However, while many studies report protein stability data, less information is found upon sugar effect on the actual encapsulation efficiency and release of proteins from polymeric particles.…”

Lactose‐containing hydrogels for enzyme stabilization

“…The release of papain from the hydrogels was observed at room temperature with a loading ratio of 1:10 (w/w) of papain to hydrogels. 10 The papainloaded hydrogels were lyophilized for the papain release assays after incubating for 24 h. A phosphate buffer solution was added to initiate the passive diffusion of papain from the hydrogels, and the release kinetics were monitored at different time intervals.…”