One-pot enzymatic synthesis of the Galα1→3Galβ1→4GlcNAc sequence within situ UDP-Gal regeneration

The trisaccharide Galα1→3Galβ1→4GlcNAcβ1→O-(CH2)8COOCH3 was enzymatically synthesized, within situ UDP-Gal regeneration. By combination in one pot of only four enzymes, namely, sucrose synthase, UDP-Glc 4′-epimerase, UDP-Gal:GlcNAc β4-galactosyltransferase and UDP-Gal:Galβ1→4GlcNAc α3-galactosyltransferase, Galα1→3Galβ1→4GlcNAcβ1→O-(CH2)8COOCH3 was formed in a 2.2 µmol ml−1 yield starting from the acceptor GlcNAcβ1→O-(CH2)8COOCH3. This is an efficient and convenient method for the synthesis of the Galα1→3Galβ1→4GlcNAc epitope which plays an important role in various biological and immunological processes.

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