Glyko® β(1-4)-Galactosidase [GKX-5014]

$ 205.00


The enzyme releases non-reducing terminal β(1-4)-linked galactose from oligosaccharides and glycoproteins [1]. This specificity is only evident at enzyme concentrations < 100mU/ml. At higher concentrations, hydrolysis of β(1-3)-linked galactose occurs.


Figure 1: Cleavage specificity for GKX-5014 β(1-4)-Galactosidase.

Streptococcus pneumoniae.

Due to its high selectivity the enzyme is an extremely useful reagent for the identification of non-reducing terminal β(1-4)-linked galactose residues. As such the enzyme has been extensively used for detailed structural analysis in conjunction with broader specificity bovine testes β-galactosidase (GKX-5013) or Jack bean β-galactosidase (GKX-5012).

Suggestions for Use:
Procedure For De-galactosylation
1. Add up to 100 μg of asialoglycoprotein or 1 nmol of oligosaccharide to tube.
2. Add de-ionized water to a total of 14 μl.
3. Add 4 μl of 5x Reaction Buffer.
4. Add 2 μl β(1-4) Galactosidase.
5. Incubate at 37 °C for 1 hour.

For glycoproteins, cleavage may be monitored by SDS-PAGE if the size differential between native and de-galactosylated protein is sufficient for detection.

For the cleavage of Galβ(1-4) linkages occurring in isolated glycans, the substrate reaction concentration should be approximately 10-20 μM. Reconstitute the enzyme in 1X Incubation buffer to give a reaction concentration of 80 mU/mL. Incubate for up to 18 hours at 37 °C.

Ships with:
WS0049 5x Reaction Buffer B [250 mM sodium phosphate (pH 6.0)]

20 mM Tris-HCl, 25 mM NaCl (pH 7.5)

Reaction Buffer:
5X concentrated buffer which when diluted gives 50 mM sodium phosphate pH 6.0.

MW: 220-247 kD

pH optimum/range:

Unit Definition:
One unit is defined as the amount of enzyme required to hydrolyze 1 μmole oNP-β-D-galactopyranoside per min at pH 6.0 and 37°C.

Size: 200 mU (100 µl)

Concentration: ≥ 2 U/ml

Product Code: GKX-5014


  1. Paulson JC, Prieels JP, Glasgow LR, Hill RL. Sialyl- and fucosyltransferases in the biosynthesis of asparaginyl-linked oligosaccharides in glycoproteins. Mutually exclusive glycosylation by beta-galactoside alpha2 goes to 6 sialyltransferase and N-acetylglucosaminide alpha1 goes to 3 fucosyltransferase.  1978 Aug 25;253(16):5617-24.


Q. Can I use this enzyme in a digestion with multiple glycosidases?

A. We use an ammonium acetate buffer when we run multi-enzyme exoglycosidase digests that include GKX-5014 β(1-4)-Galactosidase. We suggest a 10X reaction buffer of 500mM ammonium acetate pH 5.5 w/ 0.05% azide. The reaction buffer can also be used diluted to 20X (25mM ammonium acetate) and lower with purified glycans. This buffer works with most of our exoglycosidases in an overnight digestion: GK80040, GK80021, GKX-5014, GKX-5013, GKX-5023, GKX-5007, GKX-5010. Generally, 2 μl of each exoglycosidase is used in a 20μl reaction with an overnight incubation (16 hours) at 37°C. Please Contact Us for more details.

If the glycans need to purified from the reaction prior to analysis by e.g. LC-MS, samples can be spun through Nanosep® 10K Omega spin filters to remove protein. For data using these methods, please see our poster, Characterizing Low-Abundance Glycans on Therapeutic Proteins.

Also, acetate buffers work better than phosphate for injecting onto LC (phosphate phase separates in acetonitrile), and acetate is volatile so samples can be dried down if necessary.

Product Citations
  1. Tanaka-Okamoto M, Hanzawa K, Mukai M, Takahashi H, Ohue M, Miyamoto Y. Identification of internally sialylated carbohydrate tumor marker candidates, including Sda/CAD antigens, by focused glycomic analyses utilizing the substrate specificity of neuraminidase.  2018 May 1;28(5):247-260.
  2. Epp A, Hobusch J, Bartsch YC, Petry J, Lilienthal GM, Koeleman CAM, Eschweiler S, Möbs C, Hall A, Morris SC, Braumann D, Engellenner C, Bitterling J, Rahmöller J, Leliavski A, Thurmann R, Collin M, Moremen KW, Strait RT, Blanchard V, Petersen A, Gemoll T, Habermann JK, Petersen F, Nandy A, Kahlert H, Hertl M, Wuhrer M, Pfützner W, Jappe U, Finkelman FD, Ehlers M.  Sialylation of IgG antibodies inhibits IgG-mediated allergic reactions.   2017 Jul 18. pii: S0091-6749(17)31101-6. doi: 10.1016/j.jaci.2017.06.021. [Epub ahead of print]
  3. Reesink HL, Bonnevie ED, Liu S, Shurer CR, Hollander MJ, Bonassar LJ, Nixon AJ.  Galectin-3 Binds to Lubricin and Reinforces the Lubricating Boundary Layer of Articular Cartilage.   2016 May 9;6:25463.
  4. Zhai S, Xu H, Jiang X, Sun S, Qin L, Wei D, Hu L.  Expression Depression of CD300LG-γ in Human Pulmonary Carcinoma.   2016 Apr;35(2):94-9.
  5. Tanaka-Okamoto M, Yabu M, Mukai M, Takahashi H, Fujiwara Y, Ohue M, Kamada Y, Miyoshi E, Miyamoto Y.  Elevation of CA19-9-Related Novel Marker, Core 1 Sialyl Lewis A, in Sera of Adenocarcinoma Patients Verified by a SRM-Based Method.   2016 Jan 4;15(1):152-65.
  6. Dashivets T, Thomann M, Rueger P, Knaupp A, Buchner J, Schlothauer T.  Multi-Angle Effector Function Analysis of Human Monoclonal IgG Glycovariants.   2015 Dec 11;10(12):e0143520. doi: 10.1371/journal.pone.0143520. eCollection 2015.
  7. Chachadi VB, Bhat G, Cheng PW.  Glycosyltransferases involved in the synthesis of MUC-associated metastasis-promoting selectin ligands.   2015 Sep;25(9):963-75.
  8. Song T, Aldredge D, Lebrilla CB.  A Method for In-Depth Structural Annotation of Human Serum Glycans That Yields Biological Variations.   2015 Aug 4;87(15):7754-62.
  9. Thomann M, Schlothauer T, Dashivets T, Malik S, Avenal C, Bulau P, Rüger P, Reusch D. In vitro glycoengineering of IgG1 and its effect on Fc receptor binding and ADCC activity.  2015 Aug 12;10(8):e0134949. doi: 10.1371/journal.pone.0134949. eCollection 2015.
  10. Borges CR, Rehder DS.  Glycan structure of Gc Protein-derived Macrophage Activating Factor as revealed by mass spectrometry.   2016 Sep 15;606:167-79.
  11. Nguyen-Khuong T, Everest-Dass AV, Kautto L, Zhao Z, Willcox MD, Packer NH.  Glycomic characterization of basal tears and changes with diabetes and diabetic retinopathy.   2015 Mar;25(3):269-83.
  12. Sudo M, Yamaguchi Y, Späth PJ, Matsumoto-Morita K, Ong BK, Shahrizaila N, Yuki N.  Different IVIG glycoforms affect in vitro inhibition of anti-ganglioside antibody-mediated complement deposition.   2014 Sep 26;9(9):e107772. doi: 10.1371/journal.pone.0107772. eCollection 2014.
  13. Suila H, Hirvonen T, Ritamo I, Natunen S, Tuimala J, Laitinen S, Anderson H, Nystedt J, Räbinä J, Valmu L.  Extracellular o-linked N-acetylglucosamine is enriched in stem cells derived from human umbilical cord blood.   2014 Apr 1;3(2):39-44.
  14. Chevreux G, Faid V, Scohyers JM, Bihoreau N.  N-/O-glycosylation analysis of human FVIIa produced in the milk of transgenic rabbits.   2013 Dec;23(12):1531-46. 
  15. Kurz S, Jin C, Hykollari A, Gregorich D, Giomarelli B, Vasta GR, Wilson IB, Paschinger K.  Hemocytes and plasma of the eastern oyster (Crassostrea virginica) display a diverse repertoire of sulfated and blood group A-modified N-glycans.   2013 Aug 23;288(34):24410-28.
  16. Umemoto E, Tanaka T, Kanda H, Jin S, Tohya K, Otani K, Matsutani T, Matsumoto M, Ebisuno Y, Jang MH, Fukuda M, Hirata T, Miyasaka M.  Nepmucin, a novel HEV sialomucin, mediates L-selectin-dependent lymphocyte rolling and promotes lymphocyte adhesion under flow.   2006 Jun 12;203(6):1603-14.

Product Safety Documentation for GKX-5014:

Product/Part No. Description
GKX-5014 Glyko® β(1-4)-Galactosidase (Streptococcus pneumonia) 
WS0049 5X Reaction Buffer B