Beta-(1-3,4,6) Galactosidase-multizyme

Beta-(1-3,4,6) Galactosidase cleaves all β-linked, non-reducing, terminal galactose from complex carbohydrates and glycoproteins.

A mixture of two recombinant enzymes, E-BG07 ( Streptococcus pneumoniae) and E-BG06(Xanthamonos)

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Specsheet
CofA
SDS: US   EU

Part Number – Amount of Enzyme
E-BG05       – 60 µLs¹
E-BG05-20   – 20 µLs¹
E-BG05-200 – 200 µls²
¹ includes buffer
² includes enzyme only

Product Description

β-D-galactoside galactohydrolase, Exo-(1-3,4,6)-beta-D-galactanase,
 Lactase, beta-(1-3,4,6) galactosidase

Beta-(1-3,4,6) Galactosidase recombinant mixture cleaves all β(1-3), β(1-4), and β(1-6) linked non-reducing, terminal galactose. Fucose, but not sialic acid, linked to the penultimate N-acetylglucosamine will block cleavage of the galactose.

β(1-4) galactose is by far the most common linkage found in N-linked oligosaccharides. For cleavage of this linkage we recommend β(1-4) Galactasidase (E-BG07).

Source This is a mixture of two recombinant enzymes, both expressed in E. coli. It includes E-BG07, β(1-4) galactosidase, and E-BG06, a β(1-3,6) galactosidase.

Contents
60 µls of Beta-(1-3,4,6) Galactosidase in 20 mM Tris-HCl, 25 mM NaCl, pH 7.5
5x Reaction Buffer- 250 mM NaPO4, pH 5.0

The supplied buffer concentrate provides the optimal pH for enzyme activity with the standard substrate. If glycosidase treatment is performed at suboptimal pH because of glycoprotein solubility or activity requirements, expect some diminution in enzyme activity.

Specific Activity >70 U/mg
Activity >6 U/ml

Molecular weight ~65,000 and 240,000 daltons

Rxn pH 5

Suggested usage
1. Add up to 100 μg of asialoglycoprotein or 1 nmol of oligosaccharide to tube.
2. Add deionized water to 13 µl.
3. Add 4 µl 5x Reaction Buffer 5.0
4. Add 2 µL of enzyme
5. Incubate one hour at 37˚C.

Specifictity Non-reducing terminal β-linked galactose. Fucose linked to the penultimate N- acetylglucosamine will block cleavage of the galactose.

Specific Activity One unit of ß(1-3,4,6)Galactosidase is defined as the amount of enzyme required to produce 1 µmole of p-nitrophenol (pNP) in 1 minute at 37˚C, pH 4.0 from p-nitrophenyl-Beta-D-galactopyranoside.

Storage Store enzyme at 4˚C.

Purity Each lot of β(1-3,4,6) Galactosidase is tested for contaminating protease as follows: 10 μg of denatured BSA is incubated for 24 hours at 37°C with 2 μL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation.

The production host strain has been extensively tested and does not produce any detectable glycosidases.

Stability Stable at least 12 months when stored frozen.

Beta-(1-3,4,6) Galactosidase References

Guile GR, Rudd PM, Wing DR, Prime SB, Dwek RA. A rapid high resolution high-performance liquid chromatographic method for separating glycan mixtures and analyzing oligosaccharide profiles. Anal Biochem. 1996; 240(2):210-26.

Jacob GS, Scudder P. Glycosidases in structural analysis. Methods Enzymol. 1994;230:280-99.

Distler JJ, Jourdian GW. The purification and properties of beta-galactosidase from bovine testes. J Biol Chem. 1973; 248(19):6772-80.