Fructooligosaccharides (FOS) are important prebiotics synthesized by the enzymatic activity of inulosucrase, and understanding their molecular interactions is essential for exploring potential modulators. This study examined the interaction between inulosucrase from Aspergillus awamori (PDB ID: 1Y4W) and the compound EPE (4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid) using an in silico molecular docking approach. Since the 1Y4W structure lacks a co-crystallized ligand, EPE was selected as a surrogate ligand derived from PDB ID: 3SC7 and prepared for docking simulations. Docking with AutoDock Vina produced a best binding affinity of "“5.6 kcal/mol, with the strongest hydrogen bond observed with TRP305 (1.92 Ã…) and additional interactions with TYR279 (3.75 Ã…). These results indicate a moderate but stable interaction, weaker than the native substrate fructose reported in crystallographic studies. Overall, EPE shows potential as a modulatory compound of inulosucrase activity and provides a basis for further enzymological and biotechnological research.