The living factory: In vivo Production of N-acetyllactosamine containing carbohydrates in E. coli |
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Authors: | Emmanuel Bettler Eric Samain Valerie Chazalet Claude Bosso Alain Heyraud David H Joziasse Warren W Wakarchuk Anne Imberty Roberto A Geremia |
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Institution: | (1) Centre de Recherches sur les Macromolécules Végétales CNRS (Affiliated with the Joseph Fourier University), B.P. 53X, 38041 Grenoble cedex 9, France;(2) Department of Medical Chemistry, Vrije Universiteit, van der Boechstraat 7, 1081 BT Amsterdam, The Netherlands;(3) Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, K1A 0R6, Canada |
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Abstract: | Scientific and commercial interest in oligosaccharides is increasing, but their availability is limited as production relies on chemical or chemo-enzymatic synthesis. In search for a more economical, alternative procedure, we have investigated the possibility of producing specific oligosaccharides in E. coli that express the appropriate glycosyltransferases. The Azorhizobium chitin pentaose synthase NodC (a (1,4)GlcNAc-transferase), and the Neisseria (1,4)galactosyltransferase LgtB, were co-expressed in E. coli. The major oligosaccharide isolated from the recombinant strain, was subjected to LC-MS, FAB-MS and NMR analysis, and identified as Gal(1,4) GlcNAc(1,4)]4GlcNAc. High cell density culture yielded more than 1.0 gr of the hexasaccharide per liter of culture. The compound was found to be an acceptor in vitro for Gal(1,4)GlcNAc (1,3)galactosyltransferase, which suggests that the expression of additional glycosyltransferases in E. coli will allow the production of more complex oligosaccharides. |
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Keywords: | oligosaccharides glycosyltransferases glycobiology N-acetyllactosamine |
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