GalactEXO is a β-galactosidase for efficient hydrolysis of galactose residues from N- and O-linked glycans. The GalactEXO enzyme display broad activity on β1-3 and β1-4 galactose on N- or O-glycosylated proteins or free oligosaccharides. GalactEXO is available as a 2000 units lyophilized galactosidase mix for hydrolysis of galactose from 2 mg glycoprotein.
| Product Group | |
|---|---|
| Source/Origin | Cloned from A. muciniphila, Produced in E. coli (His-tagged), 87 kDa, 109 kDa |
| Product Composition | |
| Digestion Site | |
| Specificity | GalactEXO hydrolyses terminal β1-3 and β1-4 galactose on glycoproteins |
| Substrates | |
| Reaction Conditions | |
| Digestion Buffers | |
| Buffer Compatibility | Displays high activity in buffers with pH values from 5.5 to7.5 and over a wide range of ionic strength., Some optimizations might be required if a buffer other than the recommended reaction buffer is used. |
| Unit Definition | 1 unit hydrolyzes galactoses from >95 % of 0.15 nmol Gal-β1,3-GalNAc-pNP and 2 nmol Gal-β1,4-GlcNAc-pNP when incubated in 20 mM Tris pH 6.8 at 37°C for 30 min, 1 unit hydrolysis galactose from 1 ug glycoprotein |
| Application |
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.
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.
Yes, both beta1-3 linked galactose on O-glycans and beta1-4 galactose on N-glycans are hydrolyzed by 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.
