Glyko® β N-Acetylhexosaminidase [GK80050]

Price:
$ 275.00

Description

Cleaves all non-reducing terminal β-linked N-acetylglucosamine (GlcNAc). Also known as N-Acetyl-β-D-glucosaminidase, β-N-Acetylglucosaminidase. β(1-2,3,4,6)-specific. Use with GKX-5003 to determine the linkage position of β-linked GlcNAc residues. Does not release bisecting GlcNAc of N-glycans. No activity toward N-acetylgalactosamine (GalNAc).

Source:
 Recombinant gene from Streptococcus pneumoniae, expressed in E. coli.

Specificity:
This enzyme releases non-reducing terminal β(1-2,3,4, and 6)-linked N-acetylglucosamine from complex carbohydrates. When incubated with oligosaccharides at low concentrations (less than 50 mU/ml) 
the enzyme can differentiate between GlcNAcβ1-2Man, GlcNAcβ1-4Man and GlcNAcβ1-6Man linkages. Under such conditions, the enzyme cleaves essentially only β1-2 linked GlcNAc, with two provisos. Firstly, β1-2 GlcNAc is not hydrolyzed if the mannose to which it is substituted has a substitution at C-6. Thus, the enzyme is useful for the analysis of tri-antennary oligosaccharides. Secondly, if the β-linked mannose of the conserved pentasaccharide core is substituted with a "bisecting" GlcNAc then only the β1-2 linked GlcNAc linked to mannose on the α1-3 arm is cleaved. At higher concentrations of the enzyme, β(1-4) and β(1-6)-linked GlcNAc may also be hydrolyzed. The reported activity against O-linked oligosaccharides suggests that GlcNAc β1-3Gal and GlcNAc β1-6Gal are digested1.

Although the specificity towards oligosaccharides containing GalNAc is not known, the enzyme, in common with other glucosaminidases, also hydrolyses pNP-β-GalNAc at pH 6.0 and 37C. The activity towards this substrate is ~7 fold less than towards pNP-βGlcNAc.

 

Figure 1: Cleavage specificity for GK80050 β N-Acetylhexosaminidase.

Applications:
Due to the complex nature of the specificity of this enzyme, it has been used extensively for the analysis of the positional isomerism of GlcNAc in glycans. In this respect, it is complementary to the β-N-acetylhexosaminidase from Jack Bean (
GKX-5003).

Ships with:
WS0059 5x Incubation Buffer A [250 mM sodium phosphate pH 5.0]

Formulation:
20 mM Tris-HCl, 50 mM NaCl (pH 7.5)

Reaction Buffer:
5X concentrated buffer which when diluted gives 50 mM sodium phosphate pH 5.0.

pH optimum/range:
5.0

Unit Definition:
One unit is defined as the amount of enzyme required to catalyze the release of 1 μmole of p-nitrophenol from p-nitrophenol-N-acetyl-β-D-glucosaminide per minute at 37°C, pH 5.0.

Size: 1.6 U (40 μl)

Concentration: ≥ 40 U/ml

Product Code: GK80050

1. Yamashita K, Ohkura T, Yoshima H, Kobata A. Substrate specificity of Diplococcal beta-N-acetylhexosaminidase, a useful enzyme for the structural studies of complex type asparagine-linked sugar chains.  1981 May 15;100(1):226-32.

Product Citations
  1. 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.
Safety

Product Safety Documentation for GK80050:

Product/Part No. Description
GK80050 Glyko® β N-Acetylhexosaminidase 
WS0049 5x Reaction Buffer B 

 


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