An efficient method for production of uridine 5 '-diphospho-N-acetylglucosamine

Uridine 5'-diphospho-N-acetylglucosamine (UDP-GlcNAc) has been synthesized by a yeast-based method from 5'-UMP and glucosamine, in which yeast cells c atalyze the conversion of 5'-UMP to 5'-UTP and provide enzymes involved in UDP-GlcNAc synthesis using S'-UTP and glucosamine as substrates. However, t his conventional method is not suitable for practical production of UDP-Glc NAc because of the low yield of the product. We found that the yqgR gene pr oduct of Bacillus subtilis, which has been identified as a glucokinase, can catalyze the phosphorylation of N-acetylglucosamine (GlcNAc) to give GlcNA c-6-phosphate, an intermediate of UDP-GlcNAc biosynthesis. The addition of the yqgR gene product to the yeast-based reaction system enabled us to synt hesize UDP-GlcNAc using GlcNAc in place of glucosamine. The addition of two enzymes, GlcNAc-phosphate mutase and UDP-GlcNAc pyrophosphorylase, increas ed the yield of UDP-GlcNAc, Using this novel method, UDP-GlcNAc was produce d at an amount of 78 mM from 100 mM 5'-UMP and 100 mM GlcNAc.