||In recent years, supramolecular hydrogels have attracted much attention because of their potential applications in biotechnology and bioengineering. Herein, on the basis of the synthesis of poly(pseudorotaxane)-tyramine-succinate ester (PPRX-TSa), poly(rotaxane)-succinate ester (PRx-Sa) and 4-hydroxyl phenyl acetamide (CHPA), the supramolecular hydrogels were fabricated under physiological condition by enzymatic crosslinking with horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). The structures of supramolecules and modified chitosan were characterized by 1H-NMR spectra. The PPRx-CHPA and PRx-CHPA in aqueous solution were rapidly formed the hydrogel around 10 and 20 seconds respectively. The physicochemical properties of the hydrogel were measured on the difference of the HRP content and H2O2 molar ratio. Obtained results suggest that enzymatic triggered supramolecular hydrogels could be applied as an injectable material in the tissue engineering and drug delivery.