Fructosyltransferase (FTase) is a key glycosidase with hydrolytic and transglycosylation functions that can utilize sucrose to generate oligofructose (FOS), which is extremely important in the food industry as well as in plants and microorganisms. However, there remain significant gaps in our understanding of the catalytic mechanism of FTase, particularly regarding the effect of regulatory mechanisms of residues on enzyme catalytic activity. In this study, molecular dynamics simulations and immobilized enzyme catalysis experiments were employed to investigate the structural dynamics and catalytic activity of QU10-FTase. The effects of structure and activity regulation of QU10-FTase induced by different environments, including the immobilized Fe