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GalactEXO® – Hydrolysis of β1-3,4 Galactose
GalactEXO is a β-Galactosidase mix that hydrolyses β1-3,4-linked terminal galactose on N- and O-glycosylated proteins or oligosaccharides.
SmartEnzymes™
GalactEXO is applicable for trimming of galactose on released glycan structures for exoglycosidase arrays or for the generation of antibodies with homogenous G0/G0F glycan profiles.
Why GalactEXO®?
- Highly efficient β-galactosidase mixture – fast removal of terminal β1-3 and β1-4-linked galactose
- Aids identification of glycation modifications at the subunit level
- Available immobilized in ready-to-use spin columns
β-galactose on N- and O-glycosylated proteins
2 h reaction
No need for co-factors
Hydrolyzes terminal β1-3 and β1-4-linked galactose
Product Formats
GalactEXO® Immobilized
Immobilized enzyme for hydrolysis of β1-3,4-linked galactose in spin columns
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About GalactEXO®
GalactEXO is an exoglycosidase and contains a mix of β-galactosidases for efficient removal of galactose residues on N- and O-glycosylated proteins or oligosaccharides. The mix is composed of two galactosidases for highly efficient hydrolysis of both β1-3 and β1-4-linked galactoses.
GalactEXO hydrolyzes glycoproteins under native conditions and display activity in a broad pH range, 5.5 to 7.5. The enzymes in GalactEXO are derived from Akkermansia muciniphila and expressed in E. coli. GalactEXO is composed of two galactosidases, both with His-tags, and the molecular weights of the components are 87 kDa and 109 kDa.
Applications
Removal of glycan β-galactose to simplify IgG glycan heterogeneity, improve LC-MS analysis, and reveal underlying glycation modifications.
Citations
Resources
SmartEnzymes™ for the Glycosylation Charcterization of Glycoproteins
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FAQ and Support
Does GalactEXO digest fucosylated glycan structures such as Lewis X or Lewis a?
No. Branching fucose linked to the same residue as the β-galactose inhibits GalactEXO activity and needs to be removed prior to GalactEXO treatment. The FucosEXO enzyme can be used for hydrolysis of α1-2, α1-3 and α1-4 linked fucose.
Does GalactEXO hydrolyze galactose on N and O-glycosylated proteins?
Yes, both beta1-3 linked galactose on O-glycans and beta1-4 galactose on N-glycans are hydrolyzed by GalactEXO.
How long is the reaction time for GalactEXO?
For most substrates such as antibodies or Fc-fusion proteins a 2 h incubation results in hydrolysis of over 95% of the present galactose. More complex substrates with high degree of galactosylation may require longer incubation times.
Can you combine GalactEXO with SialEXO or SialEXO 23 to remove both galactoses and sialic acids in a single reaction?
Yes, you can. All three enzymes perform well in the same reaction buffer (20 mM Tris pH 6.8). There is no need to increase incubation time when combining the enzymes.
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