||Injectable alginate hydrogels offer a wide range of attractive benefits. In this study, enzyme-mediated injectable pullulan hydrogels containing blood coagulation agents such as thrombin and calcium chloride were fabricated to use as a hydrogel type hemostatic agent. The pullulan hydrogel has many advantages such as biodegradability, flexibility, and blood coagulability. The modified pullulan was characterized by ATR-FTIR, 13C NMR and 1H NMR to confirm introduction of tyramines. Gelation, rheological properties were decided as a function of concentration of horseradish peroxidase and pullulan solutions. The in vivo animal study showed that the modified hydrogel considerably reduced bleeding quantity and time compared to the unmodified model. The modified pullulan hydrogel containing blood coagulation agents has adjusted gelation time, biodegradability, hemostatic ability as an effective hemostatic agent of massive hemorrhage.