Conversion of the adenosine moieties of RNA into ATP for use in cofactor recycling

We report here a practical procedure for transforming the adenosinc moieties of RNA into ATP (Scheme I). Yeast RNA and the requis i te nzymes are commercial ly avai lable and incxpensive, and diammonium acetyl phosphate (AcP) is easily prepared.r The mixture of nucleot ides generated by th is procedure (containing ATP, GMP, UMP, CMP, and other minor const i tuents) may be used without pur i f icat ion in synthet ic schemes involving enzymatic atalysis with ATP recycling.2.l This method is the most practical one presently available for generat ing thc ATP rcquired in such schemes. Thc fol lowing detai ls i l lustrate the manipulat ions involved in prepar ing the ATP-containing mixture, and in i ts use in ATP-requir ing enzymat ic synthesis. In a rcprcscntative procedure, RNA (100 g, 85% pure, from Torula yeast, Sigma Chemical Co.) was dissolved in 400 mL of water, and the solut ion adjusted to pH 5.6 wi th NaOH and t o 0 . 1 m M i n Z n ( l l ) . 4 N u c l e a s e P r s ( 8 . C . 3 . 1 . 4 . , 0 . 2 0 m g , 9 3 U6) was added and the so lu t ion was a l lowed to s t i r a t 65 oC.