Practical Enzymatic Synthesis of Adenosine 5'-O-(3-Thiotriphosphate) (ATP-7-S)t

An enzymatic procedure for the synthesis of adenosine 5'-O-(3-thiotriphosphate) (ATP-r-S) on a 5O-mmol scale from dihydroxyacetone, sodium thiophosphate, ADP, and phosphoenol pyruvate is described. The synthesis uses polyacrylamide gel immobilized glycerokinase coupled to a pymvate kinase catalyzed ATP cofactor regeneration system, and polyacrylamide gel immobilized triosephosphate isomerase, glyceraldehyde 3-phosphate dehydrogenase, and phosphoglycerate kinase coupled to a lactate dehydrogenase catalyzed NAD cofactor regeneration system. The ATP-7-S is purified by adsorption on Dowex 1 and isolated as the sodium or barium salts in -90Vo purity.