Bettler E, Samain E, Chazalet V, Bosso C, Heyraud A, Joziasse DH, Wakarchuk WW, Imberty A, Geremia RA The living factory: In vivo production of N-acetyllactosamine containing carbohydrates in E-coli Glycoconjugate Journal16(3) (1999)
205-212
NCBI PubMed ID:10596895 Journal NLM ID:8603310 Publisher: Kluwer Academic Publishers Correspondence: geremiacermav.cnrs.fr Institutions: Centre de Recherches sur les Macromolecules Vegetales CNRS, Grenoble cedex, France
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 β(1,4)GlcNAc-oligosaccharide synthase), and the Neisseria β(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 βGal(1,4)[βGlcNAc(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 βGal(1,4)GlcNAc α(1,3)galactosyltransferase, which suggests that the expression of additional glycosyltransferases in E. coli will allow the production of more complex oligosaccharides.
carbohydrates, carbohydrate, production, N-acetyllactosamine, in vivo, in vivo production
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