Protein immobilization on solid surfaces is a prevalent strategy in biotechnology and industry, offering advantages in terms of stability and reusability. Various methods, including covalent attachment, adsorption, covalent cross-linking, and entrapment, have been developed, with challenges in adapting these techniques to enzymes due to their catalytic pockets. In a previous study [1], we introduced a method for grafting the glycoside hydrolase Neocallimastix patriciarum endo-β-1,4-xylanase (NpXyn11A) onto paramagnetic beads, preserving its biological activity. Here, we extend this approach to polymeric surfaces, specifically polyisoprene (PI) and polydimethylsiloxane (PDMS), known for their elasticity. The surfaces are plasma-activated and functionalized with maleimide functions to immobilize engineered proteins Jo and In through a thiol-maleimide click reaction. A custom device is employed to stretch the elastomer surfaces, altering the distance between immobilized enzymes. The specific activity of the immobilized xylanase, evaluated with a chromogenic substrate, mirrors that of the free enzyme in solution. However, the use of natural polysaccharides reveals activity variations based on the extent of surface stretching, providing insights into the impact of spatial proximity on enzyme dynamics. Our study contributes to overcoming limitations in traditional enzyme immobilization, offering valuable insights into the dynamics of multi-enzymatic complexes.