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Enzymatic Synthesis of Nucleoside‐5′‐ O ‐(1‐thiophosphates) and ( S P )‐Adenosine‐5′‐ O ‐(1‐thiotriphosphate)
Author(s) -
Barai Vladimir N.,
Zinchenko Anatoli I.,
Kvach Sergei V.,
Titovich Olga I.,
Rubinova Elena B.,
Kalinichenko Elena N.,
Mikhailopulo Igor A.
Publication year - 2003
Publication title -
helvetica chimica acta
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.74
H-Index - 82
eISSN - 1522-2675
pISSN - 0018-019X
DOI - 10.1002/hlca.200390233
Subject(s) - chemistry , adenosine , nucleoside , thiophosphate , phosphate , enzyme , column chromatography , stereochemistry , chromatography , biochemistry , organic chemistry
Treatment of adenosine with PSCl 3 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 m M MgCl 2 ; 28°, 8 h), followed by ion‐exchange column chromatography afforded AMP (8%), AMPS (recovered, 23%), ATP (11%), and ( S P )‐adenosine‐5′‐ O ‐(1‐thiotriphosphate) (( S P )‐ATP α S); (total yield 37%; 48% based on the consumed AMPS). For comparison of physicochemical properties, adenosine was chemically transformed into ATP α S as a mixture of the ( S P ) (53%) and (R P ) (44%) diastereoisomers.