METHOD OF RIBONUCLEOSIDE-5'-PHOSPHATES PREPARING Russian patent published in 1997 - IPC

FIELD: biochemistry. SUBSTANCE: 5'-mono-, di- and triphosphates of adenosine, uridine and cytidine were prepared by biophosphorylation of the corresponding ribonucleosides with yeast enzyme system in the presence of inorganic phosphate, the source of potassium and magnesium ions, carbohydrates and toluene. Reaction is terminated by acidification to pH = 4.5-4.8 and yeast slime is separated. Then method involves thermic treatment at pH = 3.0-5.0 at 45-70 C for 20-60 min and the formed precipitate is separated additionally. Reaction products were isolated from filtrate using strong-base anion-exchange resin by fractional elution: mono- and diphosphates were eluted with 0.02 N sodium chloride solution in 0.01 N hydrochloric acid and triphosphates were eluted with 0.5 N sodium chloride solution in 0.02 N hydrochloric acid. Reaction products were isolated from eluent by precipitation with organic solvent: acetone and ethanol cooled to (-8)-(-12) C. Then reaction product aqueous solutions were deionized by treatment with sulfocationite in H⁺-form or Trilon B at concentration 0.001-1.0%. The end product is isolated by repeated precipitation from deionized solution to excess of organic solvent indicated above. The end products were used in medicine, biology and biochemistry. EFFECT: improved method of preparing. 4 cl, 1 tbl