Preparative-scale enzymic synthesis of D-[14C]ribulose 1,5-bisphosphate.
A procedure is described to prepare uniformly labelled D-[14C]ribulose 1,5-bisphosphate enzymically from uniformly labelled D-[14C]glucose through the coupled reactions catalysed by hexokinase (EC 2.7.1.1), glucose 6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44) and 5-phosphoribulokinase (EC 2.7.1.19). All reagents utilized in the method are commercially available. The procedure is a reliable preparative-scale method for synthesizing the dibarium salt of D-[14C]ribulose 1,5-biphosphate with a specific radioactivity up to 7 mCi/mmol and a purity near 90%. The final product was free of other 14C-labelled sugars, sugar phosphate esters, Pi and nucleotides.
D001464:Barium; D002250:Carbon Radioisotopes; D005947:Glucose; D005954:Glucosephosphate Dehydrogenase; D006593:Hexokinase; D010428:Pentosephosphates / Q000138:chemical synthesis*; D010734:Phosphogluconate Dehydrogenase; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); D012274:Ribulosephosphates / Q000138:chemical synthesis*
D002250:Carbon Radioisotopes; D010428:Pentosephosphates; D012274:Ribulosephosphates; D001464:Barium; D010734:Phosphogluconate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); D006593:Hexokinase; C022390:phosphoribulokinase; D005947:Glucose