Enzymatic Synthesis of Nucleoside-5′-O-(1-thiophosphates) and (SP)-Adenosine-5′-O-(1-thiotriphosphate)

• Published in: Helvetica chimica acta 2003-08, Vol.86 (8), p.2827-2832
• Main Authors: Barai, Vladimir N., Zinchenko, Anatoli I., Kvach, Sergei V., Titovich, Olga I., Rubinova, Elena B., Kalinichenko, Elena N., Mikhailopulo, Igor A.
• Format: Article
• Language: English
• ISSN: 0018-019X; 1522-2675
• DOI: 10.1002/hlca.200390233

Treatment of adenosine with PSCl3 in trimethyl phosphate gave, after ion‐exchange chromatography, adenosine‐5′‐O‐monophosphate (AMP; 28%) and adenosine‐5′‐O‐monothiophosphate (AMPS; 48%). AMPS was studied as a thiophosphate residue donor in an enzymatic transphosphorylation with nucleoside phosphotransferase (NPase) of the whole cells of Erwinia herbicola. As exemplified by a number of natural and sugar‐ and base‐modified nucleosides, it was demonstrated that NPase of the whole cells of Erwinia herbicola catalyzes the transfer of both thiophosphate and phosphate residues with a similar efficiency. An incubation of AMPS in a phosphorylating extract of Saccharomyces cerevisiae (K‐phosphate buffer (0.3 M, pH 7.0); 3% glucose; 15 mM MgCl2; 28°, 8 h), followed by ion‐exchange column chromatography afforded AMP (8%), AMPS (recovered, 23%), ATP (11%), and (SP)‐adenosine‐5′‐O‐(1‐thiotriphosphate) ((SP)‐ATPαS); (total yield 37%; 48% based on the consumed AMPS). For comparison of physicochemical properties, adenosine was chemically transformed into ATPαS as a mixture of the (SP) (53%) and (RP) (44%) diastereoisomers.