The invention discloses a method for preparing adenosine triphosphate (ATP) with an immobilized enzyme method. The method comprises the following steps: (1) fixing an ATP production enzyme on an immobilized carrier to prepare an immobilized ATP production enzyme; (2) catalyzing by using the immobilized ATP production enzyme to prepare an adenosine triphosphate reaction solution; (3) separating the product adenosine triphosphate. According to the method disclosed by the invention, three types of novel ATP production enzymes including Ppk, Adk and Pap are adopted, and the ATP can be synthesized through only two steps of enzymatic reaction; a reaction process is simpler, a reaction is more easily controlled and the product quality is more stable; the ATP is prepared by adopting an immobilized enzyme catalysis method, and an immobilized enzyme can he continuously and repeatedly used for a plurality of times, so that the production cost is greatly reduced. Meanwhile, impurities including a lot of proteins, pigments and the like, which are introduced by utilizing yeasts, are avoided, and the product is more easily purified; an intermittent stirring reaction and enzyme reaction column continuous reaction system which is applicable to large-scale production of the ATP is established.