Facile and Selective Synthesis of Diadenosine Polyphosphates through Catalysis by Leucyl t-RNA Synthetase Coupled with ATP Regeneration

Abstract

Leucyl t-RNA synthetase from a thermophilic bacterium, Bacillus stearothermophilus, effectively catalyzed the synthesis of p¹, p⁴-di(adenosine 5'-)tetraphosphate (Ap₄A), p¹, p⁵-di(adenosine 5'-)-pentaphosphate (Ap₅A) and adenosine 5'-tetraphosphate (p₄A). In particular, when the reaction was coupled with an ATP recycling system involving thermostable acetate kinase and adenylate kinase, Ap₄A and Ap₅A were produced selectively in high yields. This reaction is selective, gives high yields and does not require protection of the functional groups of nucleotides, and also it can be carried out in an aqueous solution. This method is superior to the conventional organic synthesis and provides a practical means of the synthesizing diadenosine polyphosphates (Ap_nA), biologically important compounds.