Large scale enzymatic synthesis of oligosaccharides and a novel purification process.

Herein we report the practical chemo enzymatic synthesis of trisaccharide and derivatives of iGb3 and Gb3, and a novel purification process using immobilized yeast to remove the monosaccharide from the reaction mixture. High purity oligosaccharide compounds were achieved in large scale. This study represents a facile enzymatic synthesis of and novel purification process of oligosaccharide.

[1]  L. Leloir,et al.  The biosynthetic pathway of the asparagine-linked oligosaccharides of glycoproteins. , 1982, CRC critical reviews in biochemistry.

[2]  S. Hakomori,et al.  Characterization of a glycosphingolipid antigen defined by the monoclonal antibody MBr1 expressed in normal and neoplastic epithelial cells of human mammary gland. , 1984, The Journal of biological chemistry.

[3]  Chi-Huey Wong,et al.  Enzymes for chemical synthesis , 2001, Nature.

[4]  S. Hakomori,et al.  Glycosphingolipid antigens and cancer therapy. , 1997, Chemistry & biology.

[5]  P. Wang,et al.  Large-scale synthesis of globotriose derivatives through recombinant E. coli. , 2003, Organic & biomolecular chemistry.

[6]  A. Ljungh,et al.  Carbohydrate recognition by enterohemorrhagic Escherichia coli: characterization of a novel glycosphingolipid from cat small intestine. , 2003, Glycobiology.

[7]  A. Zatorski,et al.  A second generation synthesis of the MBr1 (globo-H) breast tumor antigen: new application of the n-pentenyl glycoside method for achieving complex carbohydrate protein linkages. , 2000, Chemistry.

[8]  Peng George Wang,et al.  Highly Efficient Chemoenzymatic Synthesis of α-Galactosyl Epitopes with a Recombinant α(1→3)-Galactosyltransferase , 1998 .

[9]  F. Compostella,et al.  A formal synthesis of 3-O-(4-methoxybenzyl)-azidosphingosine by a modified Julia olefination , 2002 .

[10]  Peng George Wang,et al.  Efficient chemoenzymatic synthesis of globotriose and its derivatives with a recombinant alpha-(1-->4)-galactosyltransferase. , 2002, Carbohydrate research.

[11]  R. Olsson,et al.  An enantiospecific synthesis of D-erythro-sphingosine from D-tartaric acid , 1993 .

[12]  G. Look,et al.  Enzyme-catalyzed oligosaccharide synthesis. , 1992, Analytical biochemistry.

[13]  E. Toone,et al.  Enzyme-catalyzed synthesis of carbohydrates. , 2000, Current Opinion in Chemical Biology.

[14]  L. Kaer,et al.  NKT cells: what's in a name? , 2004, Nature Reviews Immunology.

[15]  D. Ho,et al.  Recognition of bacterial glycosphingolipids by natural killer T cells , 2005, Nature.

[16]  D. Solter,et al.  New globoseries glycosphingolipids in human teratocarcinoma reactive with the monoclonal antibody directed to a developmentally regulated antigen, stage-specific embryonic antigen 3. , 1983, The Journal of biological chemistry.

[17]  C. Bertozzi,et al.  Glycans in cancer and inflammation — potential for therapeutics and diagnostics , 2005, Nature Reviews Drug Discovery.

[18]  P. Wang,et al.  Synthesis of α-Gal epitope derivatives with a galactosyltransferase–epimerase fusion enzyme , 2000 .

[19]  M. Palcic.  Biocatalytic synthesis of oligosaccharides. , 1999, Current opinion in biotechnology.

[20]  D. Ho,et al.  Bacterial glycolipids and analogs as antigens for CD1d-restricted NKT cells. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[21]  J Zhang,et al.  Sugar nucleotide regeneration beads (superbeads): a versatile tool for the practical synthesis of oligosaccharides. , 2001, Journal of the American Chemical Society.

[22]  Chi‐Huey Wong,et al.  Enzymes in oligosaccharide synthesis: active-domain overproduction, specificity study, and synthetic use of an .alpha.-1,2-mannosyltransferase with regeneration of GDP-Man , 1993 .

[23]  K. Duan,et al.  Production of High-content Galacto-oligosaccharide by Enzyme Catalysis and Fermentation with Kluyveromyces marxianus , 2006, Biotechnology Letters.

[24]  K. Nicolaou,et al.  Total synthesis of globotriaosylceramide (Gb3) and lysoglobotriaosylceramide (lysoGb3). , 1990, Carbohydrate research.

[25]  R. Proia,et al.  Lysosomal Glycosphingolipid Recognition by NKT Cells , 2004, Science.

[26]  Chi‐Huey Wong,et al.  A highly efficient multienzyme system for the one-step synthesis of a sialyl trisaccharide in situ generation of sialic acid and N-acetyllactosamine coupled with regeneration of UDP-glucose, UDP-galactose, and CMP-sialic acid , 1991 .