Synthesis of various phosphodiesters and phosphomonoesters with ribonuclease N.
1. 3'-Guanylyl-ethanol, 3'-guanylyl-propanol, and 3'-guanylyl-alpha-glycerol were synthesized by ribonuclease N1 [EC 3.1.4.8] using guanosine 2',3'-cyclic phosphate as a phosphate donor and various alcohols as phosphate acceptors. The yields of these phosphodiesters were 15%, 13.5%, 38.2%, respectively, with respect to phosphate donor under the optimum conditions. No phosphodiester was synthesized when 2-propanol was used as a phosphate acceptor. Thus, primary alcoholic hydroxyl groups may be regarded as the preferred phosphate acceptor. 2. 3'-Guanylyl-glucose and 3'-guanylyl-ribose were synthesized using glucose and ribose as phosphate acceptors. Under the optimum conditions, the yields of guanylyl-glucose amounted to 52.0%, while that of guanylyl-ribose was much lower. The guanylyl-glucose can be regarded as 3'-guanylyl-6-glucopyranose, based on the results of periodate oxidation. 3. Neither hydroxyamino acids (serine and threonine) nor N-acetylserinamide could be phosphorylated under the conditions used for the above phosphorylations. 4. 3'-Guanylyl-glycerol obtained as above was hydrolyzed by snake venon phosphodiesterase to produce glycerol 3-phosphate. The latter consisted of L-glycerol 3-phosphate (ca 17%) and the D-isomer (ca. 83%). Ribonuclease N1 thus catalyzes an asymmetric synthesis.
D000433:1-Propanol / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D006150:Guanine Nucleotides / Q000096:biosynthesis*; D006157:Guanosine Monophosphate / Q000096:biosynthesis*; D010755:Organophosphates / Q000096:biosynthesis*; D009943:Organophosphorus Compounds / Q000096:biosynthesis*; D006163:Ribonuclease T1 / Q000378:metabolism*; D012260:Ribonucleases / Q000378:metabolism*; D012266:Ribose / Q000378:metabolism
D006150:Guanine Nucleotides; D010755:Organophosphates; D009943:Organophosphorus Compounds; D000431:Ethanol; D012266:Ribose; D006157:Guanosine Monophosphate; D000433:1-Propanol; D012260:Ribonucleases; D006163:Ribonuclease T1; D005947:Glucose; D005990:Glycerol