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Enzymatic synthesis and carbon-13 nuclear magnetic resonance conformational studies of disaccharides containing. beta. -D-galactopyranosyl and. beta. -D-(1-/sup 13/C)Galactopyranosyl residues

Journal Article · · Biochemistry; (United States)
OSTI ID:5348148
Partially purified UDPgalactosyltransferase (EC 2.4.1.22) from bovine milk has been used to synthesize millimolar amounts of compounds such as Gal..beta..(1..-->..4)Glc, Gal..beta..(1..-->..4)GlcNAc-..beta..-hexanolamine, and Gal..beta..(1..-->..4)-GlcNAc..beta..(1..-->..4)GlcNAc. The same method has been used to prepare similar compounds containing /sup 13/C-enriched galactopyranosyl moieties. Gal..beta..(1..-->..4)GlcNAc-..beta..-hexanolamine was also synthesized in a solid-phase system in which the GlcNAc-..beta..-hexanolamine glycoside was covalently linked to agarose beads. At pH 7.0 and at 1 to 5 mM Mn/sup 2 + +/ the yields of the galactosyl saccharides are greater than 90% by using 10% excess of UDPGal donor. The use of a 90% enriched (1-/sup 13/C)galactosyl residue allowed the determination of the most abundant conformer about the galactopyranosyl-glycoside linkage by analysis of the carbon-carbon coupling constants from Cl to Gal to the C3', C4', and C5' of GlcNAc or Glc. 3 figures, 1 table.
Research Organization:
Michigan State Univ., East Lansing
OSTI ID:
5348148
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 19:3; ISSN BICHA
Country of Publication:
United States
Language:
English