Convenient syntheses of cytidine 5'-triphosphate, guanosine 5'-triphosphate, and uridine 5'-triphosphate and their use in the preparation of UDP-glucose, UDP-glucuronic acid, and GDP-mannose

Article abstract of DOI:10.1021/jo00293a030

This paper compares enzymatic and chemical methods for the synthesis of cytidine 5'-triphosphate, guanosine 5'-triphosphate, and uridine 5'-triphosphate from the corresponding nucleoside monophosphates on scales of ~10 g. These nucleoside triphosphates are important as intermediates in Leloir pathway biosyntheses of complex carbohydrates; the nucleoside monophosphates are readily available commercially. The best route to CTP is based on phosphorylation of CMP using adenylate kinase (EC 2.7.4.3); the route to GTP involves phosphorylation of GMP using guanylate kinase (EC 2.7.4.8); chemical deamination of CTP (prepared enzymatically from CMP) is the best synthesis of UTP. For the 10-200-mmol-scale reactions described in this paper, it is more convenient to prepare phosphoenolpyruvate (PEP), used in the enzymatic preparations, from D-(-)-3-phosphoglyceric acid (3-PGA) in the reaction mixture rather than to synthesize PEP in a separate chemical step. The in situ conversion of 3-PGA to PEP requires the coupled action of phosphoglycerate mutase (EC 2.7.5.3) and enolase (EC 4.2.1.11). The enzyme-catalyzed syntheses of uridine 5'-diphosphoglucose (UDP-Glc), uridine 5'-diphosphoglucuronic acid (UDP-GlcUA), and guanosine 5'-diphosphomannose (GDP-Man) illustrate the use of the nucleoside triphosphates.