Nattokinase is a kind of serine protease produced by the fermentation of Bacillus subtilis, which has strong thrombolytic effect.Compared with other thrombolytic agents, nattokinase is easy to be absorbed and has a rapid effect and good safety. Therefore, it is a new thrombolytic agent to be developed.The mechanism and physicochemical properties of nattokinase were summarized,Several methods of isolation and purification of nattokinase were introduced,Including column chromatography, ultrafiltration, extraction and affinity particle adsorption, as well as integrated separation and purification technology,The new method of isolation and purification of nattokinase was prospected to improve the development and application of Nattokinase.