Dittmer, Kayla ReataKayla ReataDittmerEckert, Kathrin MarinaKathrin MarinaEckertSetiawan, SherlianaSherlianaSetiawanKißling, Patrick AndréPatrick AndréKißlingOhde, DanielDanielOhdeSmirnova, IrinaIrinaSmirnovaLiese, AndreasAndreasLiese2026-06-292026-06-292026-06-25Industrial & Engineering Chemistry Research (in Press): (2026)https://hdl.handle.net/11420/63660Enzyme immobilization on stimuli-responsive hydrogels provides a strategy to control biocatalytic activity under fluctuating process conditions. In this study, the immobilization of formate dehydrogenase on three hydrogel carriers is investigated: pH-responsive poly(2-hydroxyethyl methacrylate-co-itaconic acid), temperature-responsive poly(N-isopropylacrylamide), and dual-responsive poly(N-isopropylacrylamide-co-itaconic acid). The hydrogels were characterized in terms of swelling behavior, mechanical properties, and morphology, confirming the responsiveness of these materials. Subsequently, the immobilization performance was evaluated across two carrier geometries (monoliths and particles) and two immobilization techniques (adsorption and covalent binding). The polymer chemistry was identified as the dominant factor controlling enzyme loading density, with the dual-responsive formulation achieving the highest loading despite moderate immobilization yields. These responsive hydrogels demonstrate strong potential as smart enzyme carriers for biocatalytic applications, particularly in continuous-flow reactor designs where improved substrate availability and increased convection can mitigate diffusion limitations and fully exploit both the catalytic performance and stimuli-responsive properties of the immobilized enzyme systems.en1520-50452026American Chemical Society (ACS)https://creativecommons.org/licenses/by/4.0/Technology::660: Chemistry; Chemical Engineering::660.6: BiotechnologyNatural Sciences and Mathematics::572: BiochemistryJournal Article10.1021/acs.iecr.6c0096110.15480/882.17372