Large-scale production of the carbohydrate portion of the sialyl–Tn epitope, α-Neup5Ac-(2→6)-d-GalpNAc, through bacterial coupling |
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Authors: | Tetsuo Endo Satoshi Koizumi Kazuhiko Tabata Shingo Kakita Akio Ozaki |
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Institution: | a Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6, Asahi-machi, Machida-shi, Tokyo 194-8533, Japan |
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Abstract: | α-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 α-(2→6)-sialyltransferase gene of Photobacterium damsela. C. ammoniagenes contributed to the production of UTP from orotic acid. α-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. |
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Keywords: | Enzymatic synthesis Glycosyltransferase Sugar nucleotide Oligosaccharide |
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