The use of immobilised glycosyltransferases in the synthesis of sialyloligosaccharides.
The CMP-sialic acids, cytidine 5'-(5-acetamido-3,5-dideoxy-beta-D-glycero-D-galacto-2-nonulopyranosy lonic acid monophosphate) (1) and cytidine 5'-(5-acetamido-9-O-acetyl-3,5-dideoxy-beta-D-glycero-D-galacto-2- nonulopyranosylonic acid monophosphate) (2) were prepared from CMP, phosphoenolpyruvate, N-acetylneuraminic acid or its 9-acetate, and a catalytic amount of ATP in the presence of immobilised pyruvate kinase, nucleoside monophosphate kinase, inorganic pyrophosphatase, and CMP-sialic acid synthetase. CMP-NeuAc (1) was used as a donor of N-acetylneuraminic acid in the reaction catalysed by immobilised porcine liver beta-D-Galp-(1----4)-alpha-D-GlcpNAc-(2----6)-sialyltransferase, alpha-D-Neup5Ac-(2----6)-beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1--- -2)-alpha-D- Man-OMe (5) was obtained on a 0.1-mmol scale by enzymic sialylation of beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----2)-alpha-D-Man-OMe (4), prepared by enzymic galactosylation of beta-D-GlcpNAc-(1----2)-alpha-D-Man-OMe (3). Likewise, using 2, alpha-D-Neup-5,9Ac2-(2----6)-beta-D-Galp-(1----4)-D-GlcpNAc (7) was obtained from N-acetyl-lactosamine (6).
Augé
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Institut de Chimie Moléculaire d'Orsay, U.R.A. C.N.R.S., Université Paris-Sud, France.
Fernandez-Fernandez
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Gautheron
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D000818:Animals; D002240:Carbohydrate Sequence; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D004800:Enzymes, Immobilized; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000096:biosynthesis*; D051381:Rats; D012799:Sialyltransferases / Q000378:metabolism*; D013552:Swine; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D004800:Enzymes, Immobilized; D009844:Oligosaccharides; C016735:sialooligosaccharides; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase