Development of an Efficient Enzymatic Process to Synthesize Fructose 1,6-Diphosphate

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This work was undertaken to develop an efficient enzymatic process to synthesize fructose l, 6-diphosphate (FDP) from glucose and ATP.The principal results were as follows :1.The rates of reactions catalyzed by Bacillus stearothermophilus glucokinase (GK), phosphoglucose isomerase (PGl), phosphofructokinase (PFK) and acetate kinase (AK) were measured under various conditions. By analyzing the data, the mechanisms and the kinetics of the reactions were clarified.2.The enzymatic synthesis of FDP from glucose and the enzymatic ATP regeneration were performed simultaneously in an ultrafiltration hollow-fiber reactor. The process consisted of three-step synthetic reactions catalyzed by GK,PGI and PFK,and the ATP regeneration reaction catalyzed by AK.The experimental results of the yield and the recycle number agreed well with the theoretical predictlons calculated on the basis of the simple analytical model.From the high recycle number and good yields obtained in the present experiment, the enzymatic synthesis of FDP from glucose and the ATP regeneration performed simultaneously in the ultrafiltration hollow-fiber reactor in the diffusion mode was proven to be usable with very good reactor performance.3.The FDP synthesized was purified, and the conditions under which crystalline FDP with a good grade can be obtained were determined.