The invention relates to the field of a biological technology, in particular to a method for preparing triphosadenine by an
immobilized enzyme method. The method particularly comprises the following steps: (1) preparing
ATP production enzyme; (2) immobilizing the
ATP production enzyme; and (3) separating products. The ATP can be synthesized by adopting three kinds of
ATP production enzyme such asadenosine
kinase (EC 2.7.1.20, Ark), adenylate
kinase (EC 2.7.4.3, Adk) and polyphosphoric acid-adenylate
phosphotransferase (EC 2.7.4.-, Pap) and only through two-step enzymatic reaction, the reaction process is simple, the reaction is easy to control and the product quality is more stable; the ATP is prepared by an
immobilized enzyme catalysis method, the
immobilized enzyme can be used continuously and repeatedly for many times, and the production cost is greatly reduced; meanwhile, introduction of a large amount of
pigment and other types of
nucleotide impurities caused by the use of yeastis avoided and purification is facilitated; and the method is suitable for producing the ATP on a large scale.