Abstract
A simplified preparative method of 2'-ribonucleotides has been devised. RNA is chemically hydrolyzed with sodium hydroxide and treated with 3'-nucleotidase of Bacillus subtilis. Then, each of four 2'-nucleotides is easily isolated from concomitant 2'-nucleotides, nucleosides and phosphoric acid, by using ion exchange resins.
Besides, the specificity of the 3'-nucleotidase was proved to be strictly restricted only to 3'-isomers of AMP, GMP, CMP and UMP. Possibility of the use of this enzyme in the determination of 3'-ribonucleotides in mixtures with other isomers was also indicated.
Molar ratio of 2'-to 3'-isomer of each nucleotide in the alkaline hydrolysate of the employed RNA is presented.
