Large-scale production of the carbohydrate portion of the sialyl-Tn epitope, alpha-Neup5Ac-(2-->6)-D-GalpNAc, through bacterial coupling.
Alpha-Neup5Ac-(2-->6)-D-GalpNAc, the carbohydrate portion of sialyl-Tn epitope of the tumor-associated carbohydrate antigen, was prepared by a whole-cell reaction through the combination of recombinant Escherichia coli strains and Corynebacterium ammoniagenes. Two recombinant E. coli strains overexpressed the CMP-Neup5Ac biosynthetic genes and the alpha-(2-->6)-sialyltransferase gene of Photobacterium damsela. C. ammoniagenes contributed to the production of UTP from orotic acid. Alpha-Neup5Ac-(2-->6)-D-GalpNAc was accumulated at 87 mM (45 g/L) after a 25-h reaction starting from orotic acid, N-acetylneuraminic acid, and 2-acetamide-2-deoxy-D-galactose.
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Tokyo, Japan. endo.tetsuo@kyowa.co.jp
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D000116:Acetylgalactosamine / Q000378:metabolism; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000096:biosynthesis* / Q000737:chemistry; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D019731:Carbon-Nitrogen Ligases / Q000235:genetics / Q000378:metabolism; D003352:Corynebacterium / Q000378:metabolism; D000939:Epitopes / Q000096:biosynthesis*; D004926:Escherichia coli / Q000378:metabolism; D007218:Industrial Microbiology; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D009963:Orotic Acid / Q000378:metabolism; D010776:Photobacterium / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012799:Sialyltransferases / Q000235:genetics / Q000378:metabolism; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase
D015295:Antigens, Tumor-Associated, Carbohydrate; D001426:Bacterial Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins; C055491:sialosyl-Tn antigen; D009963:Orotic Acid; D012799:Sialyltransferases; D019731:Carbon-Nitrogen Ligases; C019951:CTP synthetase; D019158:N-Acetylneuraminic Acid; D000116:Acetylgalactosamine; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase