One-pot enzymatic synthesis of the Gal alpha 1-->3Gal beta 1-->4GlcNAc sequence with in situ UDP-Gal regeneration.
The trisaccharide Gal alpha 1-->3Gal beta 1-->4GlcNAc beta 1-->O-(CH2)8COOCH3 was enzymatically synthesized, with in situ UDP-Gal regeneration. By combination in one pot of only four enzymes, namely, sucrose synthase, UDP-Glc 4'-epimerase, UDP-Gal:GlcNAc beta 4-galactosyltransferase and UDP-Gal:Gal beta 1-->4GlcNAc alpha 3-galactosyltransferase, Gal alpha 1-->3Gal beta 1-->4GlcNAc beta 1-->O-(CH2)8COOCH3 was formed in a 2.2 mumol ml-1 yield starting from the acceptor GlcNAc beta 1-->O-(CH2)8COOCH3. This is an efficient and convenient method for the synthesis of the Gal alpha 1-->3Gal beta 1-->4GlcNAc epitope which pays an important role in various biological and immunological processes.
Hokke
C H
CH
Department of Medical Chemistry, Vrije Universiteit, Amsterdam, The Netherlands.
Zervosen
A
A
Elling
L
L
Joziasse
D H
DH
van den Eijnden
D H
DH
D000818:Animals; D002240:Carbohydrate Sequence; D002417:Cattle; D004795:Enzyme Stability; D005700:Galactosyltransferases / Q000737:chemistry*; D005964:Glucosyltransferases / Q000737:chemistry*; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000138:chemical synthesis*; D014534:UDPglucose 4-Epimerase / Q000737:chemistry*; D014531:Uridine Diphosphate Galactose / Q000737:chemistry*
D009844:Oligosaccharides; D014531:Uridine Diphosphate Galactose; D005700:Galactosyltransferases; D005964:Glucosyltransferases; C022699:sucrose synthase; D014534:UDPglucose 4-Epimerase