ac-cleavage-galexo-04-outline hydrolysis of galactose

GalactEXO

GalactEXO is a β-galactosidase for efficient hydrolysis of galactose residues from N- and O-linked glycans. The GalactEXO galactosidase display broad activity on β1-3 and β1-4 galactose on N- or O-glycosylated proteins or free oligosaccharides.

GalactEXO is available in two different product formats:

  • GalactEXO - lyophilized β-galactosidase mix for complete hydrolysis of galactose from 2 mg glycoprotein in 2h.
  • Immobilized GalactEXO - immobilized GalactEXO contains microspin columns for fast and efficient hydrolysis of galactose from 0.5 mg glycoprotein

Key Characteristics

  • Galactosidase hydrolyzing galactose on N- and O-glycoproteins or free oligosaccharides
  • β1-3 and β1-4 linked galactose residues
  • 2 h reaction
  • Requires no co-factors

GalactEXO Enzyme

GalactEXO is 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 molecular weights of the components are 87 kDa and 109 kDa, respectively.

 

Unit Definition

One unit of GalactEXO 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 will hydrolyze the galactose of 1 ug of glycoprotein.

 

Content and Storage

GalactEXO is supplied lyophilized in TBS pH 7.6, with no preservatives added.
GalactEXO is shipped cold and should be stored at -20 °C upon arrival.
After reconstitution GalactEXO is stable for 1 month at +4-8 °C.

Documents

Instructions

Product Specification

Safety Data Sheet

Certificate of Analysis

Applications

Immobilized GalactEXO

Immobilized GalactEXO is a resin with a mixture of two β-galactosidases covalently coupled to agarose beads for efficient removal of galactose residues (β1-3 and β1-4 linked) on N- and O-glycosylated proteins. The enzymes in GalactEXO are derived from Akkermansia muciniphila and expressed in E. coli. Degalactosylated proteins are generated without the enzymes in the final preparation. The glycoprotein sample is incubated with Immobilized GalactEXO and the hydrolyzed glycoproteins are then easily collected by a centrifugation step. The recommended buffer for immobilized GalactEXO is 20 mM Tris pH 6.8. The protocol may need optimization regarding buffer compatibility and incubation time for individual glycoproteins.

 

Unit Definition

One column of Immobilized GalactEXO 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, monitored by reversed phase chromatography.

 

Content and Storage

Immobilized GalactEXO Microspin columns contain sufficient material each to remove galactoses from 0.5 mg glycoprotein. The resin is supplied in 20% EtOH with no preservatives added.
Immobilized GalactEXO is shipped cold and should be stored at +4-8°C upon arrival.

  • G1-GM6-025

    Immobilized GalactEXO Microspin, Digest 5 x 0.5 mg

    G1-GM6-025795.00Add to cart
  • G1-GM6-050

    Immobilized GalactEXO Microspin, Digest 10 x 0.5 mg

    G1-GM6-0501,295.00Add to cart

Documents

Instructions

Product Specification

Safety Data Sheet

Certificate of Analysis

Applications

FAQ

Popular FAQ

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.

Yes, both beta1-3 linked galactose on O-glycans and beta1-4 galactose on N-glycans are hydrolyzed by GalactEXO.

No. Branching fucose linked to the same residue as the β-galactose inhibits GalactEXO activity and needs to be removed prior to GalactEXO treatment.

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GalactEXO Brochure

Removal of galactose residues using GalactEXO – a broad acting β-galactosidase

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For more information about commercial rights, please contact Genovis team at info@genovis.com.

Genovis products are intended for research use only. They are not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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