title: Large-scale production of CMP-NeuAc and sialylated oligosaccharides through bacterial coupling.
abstract: A large-scale production system of cytidine 5'monophospho-N-acetylneuraminic acid (CMP-NeuAc) and sialyloligosaccharides was established by a whole-cell reaction through the combination of recombinant Escherichia coli strains and Corynebacterium ammonia-genes. For the production of CMP-NeuAc, two recombinant E. coli strains were generated that overexpressed the genes of CMP-NeuAc synthetase and CTP synthetase, respectively. C. ammoniagenes contributed to the formation of UTP from orotic acid. CMP-NeuAc was accumulated at 27 mM (17 g/l) after a 27-h reaction starting with orotic acid and N-acetylneuraminic acid. When E. coli cells that overexpressed the alpha-(2-->3)-sialyltransferase gene of Neisseria gonorrhoeae were put into the CMP-NeuAc production system, 3'-sialyllactose was accumulated at 52 mM (33 g/l) after an 11-h reaction starting with orotic acid, N-acetylneuraminic acid, and lactose. Almost no oligosaccharide byproducts other than 3'-sialyllactose were observed after the reaction. The production of 3'-sialyllactose at a 5-l jar fermenter scale was almost the same as that at a beaker scale, which indicated the high potential of the 3'-sialyllactose production on an industrial scale.
authors.affiliation: Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Machida, Japan. endo.tetsuo@kyowa.co.jp
authors.forename: T
authors.initials: T
authors.lastname: Endo
authors.forename: S
authors.initials: S
authors.lastname: Koizumi
authors.forename: K
authors.initials: K
authors.lastname: Tabata
authors.forename: A
authors.initials: A
authors.lastname: Ozaki
mesh_terms: D001709:Biotechnology / Q000379:methods; D003352:Corynebacterium / Q000378:metabolism*; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D007785:Lactose / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012794:Sialic Acids / Q000378:metabolism; D012799:Sialyltransferases / Q000235:genetics / Q000378:metabolism
chemical_list: D009844:Oligosaccharides; D011994:Recombinant Proteins; D012794:Sialic Acids; C016735:sialooligosaccharides; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid; D012799:Sialyltransferases; D007785:Lactose