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 catalyze the conversion of 5'-UMP to 5'-UTP and provide enzymes involved in UDP-GlcNAc synthesis using 5'-UTP and glucosamine as substrates. However, this conventional method is not suitable for practical production of UDP-GlcNAc because of the low yield of the product. We found that the yqgR gene product of Bacillus subtilis, which has been identified as a glucokinase, can catalyze the phosphorylation of N-acetylglucosamine (GlcNAc) to give GlcNAc-6-phosphate, an intermediate of UDP-GlcNAc biosynthesis. The addition of the yqgR gene product to the yeast-based reaction system enabled us to synthesize UDP-GlcNAc using GlcNAc in place of glucosamine. The addition of two enzymes, GlcNAc-phosphate mutase and UDP-GlcNAc pyrophosphorylase, increased the yield of UDP-GlcNAc. Using this novel method, UDP-GlcNAc was produced at an amount of 78 mM from 100 mM 5'-UMP and 100 mM GlcNAc.
Okuyama
K
K
Biochemicals Division, Yamasa Corporation, Choshi, Chiba, Japan. okuyama@yamasa.com
Hamamoto
T
T
Ishige
K
K
Takenouchi
K
K
Noguchi
T
T
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004274:DNA, Recombinant; D005941:Glucokinase / Q000378:metabolism; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000235:genetics*
D017931:DNA Primers; D004274:DNA, Recombinant; D014537:Uridine Diphosphate N-Acetylglucosamine; D005941:Glucokinase