Continuous regeneration of ATP in enzyme membrane reactor for enzymatic syntheses.

This work shows that the enzyme membrane reactor offers the opportunity to carry out the enzymatic regeneration of ATP providing continuous operation with high performance. In this system, the coenzyme is immobilized on a water-soluble polymer. These high-molecular weight derivates are entrapped within an ultrafiltration membrane together with the enzymes for production of regeneration. Several polymer derivatives of ATP have been prepared for this immobilization technique. Coenzymatic activity of these derivatives was studied with several enzymes for both ATP-requiring and ATP-regenerating reactions. PEG-N6-aminohexyl-ATP was selected as the appropriate coenzyme for operating the enzyme membrane reactor. Acetate kinase was the only enzyme providing enough activity for regeneration. Production of glucose-6-phosphate is cited as the first example. The kinetics of acetate kinase and hexokinase were examined and used to choose the operating conditions of the process. The process operated continuously for more than 1 month. With a mean conversion of 80%, the space-time yield amounted to 348 g glucose-6-phosphate/L d. The cycle number of ATP was estimated as 20, 000 mol/mol. With the continuous production of gamma-glutamylcysteine and NADP, the feasibility of the system was proven.