Study On Adenosine Triphosphate Separation And Purification Process In Biosynthesis Of ATP

In the production process of adenosine triphosphate (ATP), the method of synthesis, test, separation and purification ATP play an important role. In this paper, the method of ATP biosynthesis has been discussed with theory. The analytic technique and separation and purification technique has been studied with experiment.The biosynthesis of ATP by immobilized enzyme/cells have many advantages, such as high efficiency of reactor, low costing of main raw materials, high yield and so on. So it has been developed in recent years and become the major production method of ATP. In this method, the key techniques are the appropriate enzyme/cells of ATP biosynthesis, the appropriate method of immobilization, the appropriate reactor & operational conditions. The developing trend about it was discussed, and it has advised to the research in the future.In the process of preparation and storage, it is easy to intermingle or creation impurity as AMP, ADP. Because AMP, ADP and ATP has similar structure and property, how to separate or selective detection them is akey technique in the analytic procedure of ATP. A chromatogram method has been founded to assay ATP with conditions as below: chromatographic column(6 X 500mm, D201 resin), mobile phase ( acetic acid buffer solution, pH3.0), detection wavelength 254nm, velocity of flow 10ml/min, column temperature 23 2 C. In the range of 0.1~8g/L, it has well linear relationship between the concentration of ATP and peak area , r=0.9977. Average recovery is 98.6%. The detection process can be finished in 30min. The method has well intra-laboratory reproducibility, and can be used to separate and detect ADP or AMP.From various ion-exchange resins, a kind of macroporous anion-exchange resin D201 was selected to separate ATP. The ion-exchange isotherm of ATP on D201 resin was measured. Types of resin and the choice of washing conditions in the process of ATP separation were investigated in detail. Furthermore, the ion-exchange rate curves and the break-through curves of ATP solution in ion-exchange column were measured. In the process of separate ATP, the D201 resin with Cl- type has high adsorption capacity. It is appropriate with pH3.0, 1mol/L NaCl solution to elute ATP. The higher ATP load on resin is benefit for ATP decoration and concentration. Adsorbance of ATP on D201 resin was up to 0.326g/g (resin) and the yield for ATP in whole process was 92.1%.From the study on ATP dissolution property in acetone-water andalcohol-water, process conditions of ATP crystallization can be acquired. In the process conditions as alcohol-water(3:1, V/V), 10 C, 40g/L ATP solution, the yield of ATP crystallization is 95%.Finally, the costing and economic balance of producing ATP has been evaluated. The developing trend about it was discussed.