Improved synthesis of CMP-sialates using enzymes from frog liver and equine submandibular gland

An efficient method for the preparation of CMP-N-acetylneuraminic acid using crude or partially purified CTP:N-acylneuraminate cytidylyltransferase from equine submandibular gland or frog liver is described. The yield of the sugar nucleotide after purification by ion-exchange chromatography and gel filtration was 95%. The compound was studied by 360 MHz 1H NMR spectroscopy in addition to the usual chemical and physical analyses. The preparation of radioactive or unlabelled CMP 4-O-methyl-N-acetylneuraminic acid, which is not known to occur in nature, was achieved in 17% yield with the aid of the equine enzyme.