One-pot enzymatic production of beta-D-galactopyranosyl-(1-->3)-2-acetamido-2-deoxy-D-galactose (galacto-N-biose) from sucrose and 2-acetamido-2-deoxy-D-galactose (N-acetylgalactosamine).
Beta-D-Galactopyranosyl-(1-->3)-2-acetamido-2-deoxy-D-galactose (galacto-N-biose, GNB) is an important core structure in functional sugar chains such as T-antigen disaccharide and the core 1 sugar chain in mucin glycoproteins. We successfully developed a one-pot enzymatic production of GNB from sucrose and GalNAc by the concomitant action of four enzymes: sucrose phosphorylase, UDP-glucose-hexose 1-phosphate uridylyltransferase, UDP-glucose 4-epimerase, and galacto-N-biose/lacto-N-biose I phosphorylase in the presence of UDP-glucose and phosphate, by modifying the method of lacto-N-biose I production [Nishimoto, M.; Kitaoka, M., Biosci. Biotechnol. Biochem., 2007, 71, 2101-2104]. The reaction yield of GNB was 88% from GalNAc. GNB was isolated from the reaction mixture by crystallization after yeast treatment to obtain approximately 45 g of GNB in 95% purity from a 280-mL reaction mixture.
Nishimoto
Mamoru
M
National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Kitaoka
Motomitsu
M
D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000737:chemistry; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000096:biosynthesis / Q000737:chemistry*; D004186:Disaccharidases / Q000138:chemical synthesis*; D004187:Disaccharides / Q000138:chemical synthesis*; D004798:Enzymes / Q000378:metabolism*; D005964:Glucosyltransferases; D006005:Phosphorylases; D013395:Sucrose / Q000737:chemistry*; D014534:UDPglucose 4-Epimerase; D005694:UTP-Hexose-1-Phosphate Uridylyltransferase
D015295:Antigens, Tumor-Associated, Carbohydrate; D004187:Disaccharides; D004798:Enzymes; C000616516:galacto-N-biose; C016467:Thomsen-Friedenreich antigen; C033398:galactosyl-1,3-N-acetylglucosamine; D013395:Sucrose; D005964:Glucosyltransferases; D006005:Phosphorylases; C025901:sucrose phosphorylase; D005694:UTP-Hexose-1-Phosphate Uridylyltransferase; D004186:Disaccharidases; D014534:UDPglucose 4-Epimerase; D000117:Acetylglucosamine