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α(1-3,4,6)-Galactosidase (coffee bean), (formerly ProZyme) 5 U. Releases nonreducing terminal α(1-3,4,6)-linked galactose from oligosaccharides. Includes 5x Reaction Buffer [500 mM sodium citrate/phosphate (pH 6.0)] which when diluted gives 100 mM sodium citrate-phosphate pH 6.0.

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Enzyme Specificity
  • Coffee bean a-galactosidase cleaves nonreducing terminal galactose residues linked a1-3, a1-4 and a1-6 to polysaccharides, glycoproteins and glycans. The precise specificity of the enzyme is dependent on the nature of the glycoconjugate.  The enzyme is stable. At 37 °C in pH 6.0 buffer, full activity is recovered after 19 hours and 70% after 48 hours. Note: The enzyme has a slow turnover. After 18 hours at 37 °C with 5 U/ml enzyme, a 40 µM solution of asialo, biantennary, fucosylated oligosaccharide with outer arm α(1-3)-galactose is 67% de-α-galactosylated.
Enzyme Applications
  • Terminal a-linked galactose residues have been reported to occur on several glycoproteins and are of increasing interest. The galactose-a-1-3Gal group is widely found in glycoconjugates from non-primate mammals and New World monkeys although it is absent from humans and Old World monkeys. The potential presence of this modification and its associated antigenicity is an important consideration in the expression of recombinant glycoproteins by mammalian cell cultures. Both a(1-3)-linked galactose and the less abundant a(1-6)-linked galactose are cleaved by this enzyme. Other workers have used the enzyme for the modification of blood group specificity. The enzyme also has diagnostic applications for the determination of raffinose in food.
Enzyme Unit Definition
  • One unit is defined as the amount of enzyme required to hydrolyze 1 µmole of pNP-α-D-galactopyranoside per minute at pH 6.5 and 37oC.
Enzyme Formulation
  • Lyophilized from 100 mM sodium phosphate pH 6.5, containing 0.25 mg/ml bovine serum albumin. See note above on BSA amount.
Enzyme Source
  • Green coffee bean. Highly purified; β-galactosidase contaminating activity is not detectable, as determined by extended incubation with 2-AB NA2 (G2).
  • 5 U
Molecular Weight
  • ~26 kD