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Large-scale production of CMP-NeuAc and sialylated oligosaccharides through bacterial coupling

Large-scale production of CMP-NeuAc and sialylated oligosaccharides through bacterial coupling 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 ammoniagenes. 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 α-(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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied Microbiology and Biotechnology Springer Journals

Large-scale production of CMP-NeuAc and sialylated oligosaccharides through bacterial coupling

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References (20)

Publisher
Springer Journals
Copyright
Copyright © 2000 by Springer-Verlag Berlin Heidelberg
Subject
Life Sciences; Microbiology; Microbial Genetics and Genomics; Biotechnology
ISSN
0175-7598
eISSN
1432-0614
DOI
10.1007/s002530050017
Publisher site
See Article on Publisher Site

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 ammoniagenes. 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 α-(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.

Journal

Applied Microbiology and Biotechnology – Springer Journals

Published: Mar 13, 2000

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