SmartEnzymes GalactEXO workflow

Hydrolysis of β1-3,4 Galactose

GalactEXO is a β-Galactosidase mix that hydrolyses β1-3,4-linked terminal galactose in N- and O-glycosylated proteins.

The enzyme 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.
 

  • 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
Hydrolyzes terminal β1-3 and β1-4-linked galactose
2 h reaction
No need for co-factors

GalactEXO Lyophilized

Lyophilized enzyme for hydrolysis of β1-3,4-linked galactose

GalactEXO Immobilized

Immobilized enzyme for hydrolysis of β1-3,4-linked galactose in spin columns

About GalactEXO

GalactEXO workflow

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.

GalactEXO™ Lyophilized

Lyophilized enzyme for hydrolysis of β1-3,4-linked galactose on glycoproteins.

GalactEXO Lyophilized is available as a lyophilized powder in 2000 unit vials for hydrolysis of β1-3,4-linked galactose on 2 mg glycoprotein.
 

  • Flexible format for method development
  • Possible to combine enzymes for improved efficiency
  • Amenable to automated workflows
  • Ready to use – just add water!

Available Products

GalactEXO Lyophilized 2000 unitsLyophilized enzyme for hydrolysis of β1-3,4-linked galactose on 2 mg glycoproteinG1-GM1-020904.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One unit of GalactEXO Lyophilized 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 of GalactEXO Lyophilized will hydrolyze the galactose of 1 μg of glycoprotein.

Content and Storage

GalactEXO Lyophilized is reconstituted by addition of water. No preservatives added. After reconstitution, GalactEXO Lyophilized is stable for 1 month at +4-8°C. The product is shipped cold and should be stored at -20°C upon arrival.

Instructions

Product Specification

Safety Data Sheet

Certificate of Analysis

GalactEXO™ Immobilized

Immobilized enzyme for hydrolysis of β1-3,4-linked galactose on glycoproteins in spin columns.

The GalactEXO Immobilized spin columns contain GalactEXO enzyme covalently coupled to agarose beads, for hydrolysis of β1-3,4-linked galactose on glycoproteins without contaminating the final preparation with enzyme.

The Microspin columns are available for hydrolysis of 5 or 10 x 0.5 mg glycoprotein.

 

  • Easy-to-use spin columns
  • Increased enzyme-to-substrate ratio for improved digestion efficiency
  • No enzyme in the final sample

Available Products

GalactEXO Immobilized Microspin 5 x 0.5 mgImmobilized enzyme for hydrolysis of β1-3,4-linked galactose on 5 x 0.5 mg glycoprotein in spin columnsG1-GM6-025959.00Buy / Request a Quote
GalactEXO Immobilized Microspin 10 x 0.5 mgImmobilized enzyme for hydrolysis of β1-3,4-linked galactose on 10 x 0.5 mg glycoprotein in spin columnsG1-GM6-0501,566.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Activity

One column of GalactEXO Immobilized catalyzes the hydrolysis of galactoses from ≥ 95% of 41 μmol 4-nitrophenyl-β-D-galactopyranoside when incubated in 20 mM Tris, 10% ethanol pH 6.8 at room temperature for 15 minutes, as monitored by reversed-phase chromatography.

One Microspin column contains sufficient GalactEXO-coupled agarose beads to hydrolyze β1-3,4-linked galactose on 0.5 mg glycoprotein.

Content and Storage

GalactEXO Immobilized is supplied in 20% ethanol, shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the product!

Instructions

Product Specification

Safety Data Sheet

Certificate of Analysis

FAQ and Support

Popular FAQ

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.

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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