3001-34-1Relevant academic research and scientific papers
32. Biosynthetic production of [N2,1,3,7,9-15N]guanosine and [1,3,7,9-15N]inosine and conversion into [N6,1,3,7,9-15N]adenosine for structure elucidation of RNA by heteronuclear NMR
Niemann,Meyer,Engeloch,Botta,Hadener,Strazewski
, p. 421 - 439 (1995)
A procedure was developed for the biosynthetic preparation of 15N-labelled guanosine and inosine through the action of a mutant Bacillus subtilis strain. Crude [N2,1,3,7,9-15N]guanosine and [1,3,7,9-15N]inosine were isolated from the culture filtrate by precipitation and anion-exchange chromatography. No cell lysis and no enzymatic degradation was necessary. The per-isobutyrylated derivatives 1 and 2 were isolated from a complex mixture, purified by virtue of their different lipophilicity, and separated in three steps involving normal- and reversed-phase silica-gel chromatography. One litre of complex nutrient medium yielded 8.44 mmol of guanosine derivative and 2.84 mmol of inosine derivative with high average 15N enrichment (83.5 and 91.9 atom-%, resp.). [N6,1,3,7,9-15N]Adenosine (4) was obtained from 2',3',5'-tri-O-isobutyryl[1,3,7,9-15N]inosine (1) through the ammonolysis of its 1,2,4-triazolyl derivative with aqueous 15NH3.
