Study of hydrogel-entrapped alcohol dehydrogenase by contrast-variation SANS

Enzyme immobilisation on solid supports enhances stability and reusability, crucial for industrial applications. Our team aims to develop a novel biocatalytic flow reactor using 3D-printed enzyme-containing geometrical structures for process intensification. We use polymeric cross-linkable water-soluble resins forming an insoluble hydrogel in the 3D-printing process. The enzyme alcohol dehydrogenase (ADH) is added to the resin and physically-entrapped within the gel structures. The enzymatic activity varies with hydrogel properties, due to diffusion limitation and/or enzyme deactivation in the cross-linking process. To improve the activity of ADH, we need to determine the impact of hydrogel entrapment on ADH structure, and thus on its functionality. We will use Contrast Variation Small-Angle Neutron Scattering to study the ADH-hydrogel system. Preliminary data from Xpress beamtime on Sans2d showed promising results for soluble and hydrogel-entrapped ADH. With this proposal, three hydrogel-ADH systems will be investigated to determine the contrast match point of polymers and measure hydrogel-ADH systems at various contrasts. Data will be analysed using shape-independent fitting models and atomistic modelling. We request three days of beamtime on Sans2d to cover the necessary q-range for polymer structure determination. This study will elucidate the impact of polymer architecture on ADH structure, guiding further material development.