Chemoenzymatic synthesis of GDP-I- fucose and the Lewis X glycan derivatives

• Published in: Proceedings of the National Academy of Sciences - PNAS 2009-09, Vol.106 (38), p.16096
• Main Authors: Wang, Wei, Hu, Tianshun, Frantom, Patrick A, Zheng, Tianqing, Gerwe, Brian, del Amo, David Soriano, Garret, Sarah, Seidel, Ronald D, Wu, Peng
• Format: Article
• Language: English
• Subjects: Biochemistry; Cells; Chemical synthesis; Enzyme kinetics; Enzymes
• ISSN: 0027-8424; 1091-6490

Lewis X (Le...)-containing glycans play important roles in numerous cellular processes. However, the absence of robust, facile, and cost-effective methods for the synthesis of Le... and its structurally related analogs has severely hampered the elucidation of the specific functions of these glycan epitopes. Here we demonstrate that chemically defined guanidine 5'-diphosphate-β-l-fucose (GDP-fucose), the universal fucosyl donor, the Le... trisaccharide, and their C-5 substituted derivatives can be synthesized on preparative scales, using a chemoenzymatic approach. This method exploits l-fucokinase/GDP-fucose pyrophosphorylase (FKP), a bifunctional enzyme isolated from Bacteroides fragilis 9343, which converts l-fucose into GDP-fucose via a fucose-1-phosphate (Fuc-1-P) intermediate. Combining the activities of FKP and a Helicobacter pylori α1,3 fucosyltransferase, we prepared a library of Le... trisaccharide glycans bearing a wide variety of functional groups at the fucose C-5 position. These neoglycoconjugates will be invaluable tools for studying Le...-mediated biological processes. (ProQuest: ... denotes formulae/symbols omitted.)