Development and performance of a 3D-printable poly(ethylene glycol) diacrylate hydrogel suitable for enzyme entrapment and long-term biocatalytic applications.

Physical entrapment of enzymes within a porous matrix is a fast and gentle process to immobilize biocatalysts to enable their recycling and long-term use. This study introduces the development of a biocompatible 3D-printing material suitable for enzyme entrapment, while having good rheological and UV-hardening properties. Three different viscosity-enhancing additives have been tested in combination with a poly(ethylene glycol) diacrylate-based hydrogel system. The addition of polyxanthan or hectorite clay particles results in hydrogels that degrade over hours or days, releasing entrapped compounds. In contrast, the addition of nanometer-sized silicate particles ensures processability while preventing disintegration of the hydrogel. Lattice structures with a total height of 6 mm consisting of 40 layers were 3D-printed with all materials and characterized by image analysis. Rheological measurements identified a shear stress window of 200 < τ < 500 Pa at shear rates of 25 s-1 and 25°C for well-defined geometries with an extrusion-based printhead. Enzymes immobilized in these long-term stable hydrogel structures retained an effective activity of approximately 10% compared to the free enzyme in solution. It could be shown that the reduction of effective activity is not caused by a significant reduction of the intrinsic enzyme activity but by mass transfer limitations within the printed hydrogel structures.