Stabilizing Enzymes within Polymersomes by Coencapsulation of Trehalose

Abstract

Enzymes are essential biocatalysts and very attractive as therapeutics. However, their functionality is strictly related to their stability, which is significantly affected by the environmental changes occurring during their usage or long-term storage. Therefore, maintaining the activity of enzymes is essential when they are exposed to high temperature during usage, or when they are stored for extended periods of time. Here, we stabilize and protect enzymes by co-encapsulating them with trehalose into polymersomes. The anhydrobiotic disaccharide preserved up to about 81% of enzyme's original activity when laccase/trehalose-loaded nanoreactors were kept desiccated for 2 months at room temperature, and 75% of its activity when heated at 50 °C for 3 weeks. Moreover, the applicability of laccase/trehalose-loaded nanoreactors as catalysts for bleaching of the textile dyes Orange G, Toluidine Blue O and Indigo was proven. Our results demonstrate the advantages of co-encapsulating trehalose within polymersomes to stabilize enzymes in dehydrated state for extended periods of time, preserving their activity even when heated to elevated temperature.

Polymeric Nanoreactors as Emerging Nanoplatforms for Cancer Precise Nanomedicine

“…Interestingly, it was found that natural enzymes in polymeric nanoreactors could be highly stabilized by co‐encapsulation with trehalose. [ 33 ] For this case, even though laccase‐loaded polymeric nanoreactors were incubated at 50 °C for three weeks, 75% original activity of laccase was still maintained under the protection of trehalose.…”

The Core–Shell Structure, Not Sugar, Drives the Thermal Stabilization of Single-Enzyme Nanoparticles

“…However, in some cases, the enzyme encapsulation efficiency is relatively low, making the process uneconomical. 34 Moreover, when embedded in a polymer matrix, the diffusion of the substrate is restricted, resulting in overall decreased activity. 35 The mechanism by which trehalose protects proteins is still under debate, but the most popular hypotheses are as follows:…”

“…49,50 This can reduce the incidence of solute crystallization during storage, which is often held responsible for the damages on the protein. 34,51 However, trehalose does not always play a dramatic role in enzyme protection. For example, in the stabilization of β-galactosidase against lyophilization, trehalose was found to have a similar protective effect as poly(ethylene glycol) (PEG), 28 which is known for its disappointing stabilizing ability.…”

“…The covalent attachment of trehalose glycopolymers to proteins could further increase their stabilizing effect, presumably due to the increased local concentration of trehalose close to the enzyme. However, the modification of proteins through covalent conjugation can at the same time induce activity loss. , To reduce the initial enzyme activity loss caused by surface modification, enzymes have been loaded into either trehalose hydrogels, matrices, or polymersomes with trehalose additives without the need for a conjugation, which simplifies the synthesis and stabilization process. However, in some cases, the enzyme encapsulation efficiency is relatively low, making the process uneconomical .…”

Bioinspired Macromolecular Materials

“…Next, Nico Bruns reports on polymersome encapsulation of trehalose alongside enzymes in order to stabilize enzyme activity. 17 In this way, functional enzyme activity is maintained in spite of prolonged periods of time under desiccation as well as upon exposure to elevated heats. Julie Champion then describes work to create vesicles from recombinant fusion proteins containing elastin-like polypeptides.…”