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Immobilisation of ?-galactosidase within a lipid sponge phase: structure, stability and kinetics characterisation

Gilbert, Jennifer; Valldeperas, Maria; Dhayal, Surender K.; Barauskas, Justas; Dicko, Cedric; Nylander, Tommy

Nanoscale, 2019, vol 11, 44, pp. 21291-21301

DOI:10.1039/C9NR06675F

Abstract

In the formulation of an active enzyme enclosed in a matrix for controlled delivery, it is a challenge to achieve a high protein load and to ensure high activity of the protein. For the first time to our knowledge, we report the use of a highly swollen lipid sponge (L3) phase for encapsulation of the large active enzyme, ?-galactosidase (?-gal, 238 kDa). This enzyme has large relevance for applications in, e.g. the production of lactose free milk products. The formulation consisted of diglycerol monooleate (DGMO), and a mixture of mono-, di- and triglycerides (Capmul GMO-50) stabilised by polysorbate 80 (P80). The advantage of this type of matrix is that it can be produced on a large scale with a fairly simple and mild process as the system is in practice self-dispersing, yet it has a well-defined internal nano-structure. Minor effects on the sponge phase structure due to the inclusion of the enzyme were observed using small angle X-ray scattering (SAXS). The effect of encapsulation on the enzymatic activity and kinetic characteristics of ?-galactosidase activity was also investigated and can be related to the enzyme stability and confinement within the lipid matrix. The encapsulated ?-galactosidase maintained its activity for a significantly longer time when compared to the free solution at the same temperature. Differences in the particle size and charge of sponge-like nanoparticles (L3-NPs) with and without the enzyme were analysed by dynamic light scattering (DLS) and zeta-potential measurements. Moreover, all the initial ?-galactosidase was encapsulated within L3-NPs as revealed by size exclusion chromatography.

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