Species : |
Xanthamonas manihotis |
Description : |
α(1-3,4) Fucosidase cleaves branched non-reducing terminal fucose, linked α(1-3) or α(1-4) to the N-acetylglucosamine of terminal Gal-GlcNAc disaccharide structures. The presence of sialic acid (but not fucose) linked to the galactose will block cleavage. |
Form : |
Sterile-filtered solution |
EC : |
3.2.1.51 |
Specificity : |
Non-reducing terminal branched fucose when linked α(1-3) or α(1-4) to GlcNAc of a Gal-GlcNAc disaccharide structure. The presence of sialic acid (but not fucose) linked to the galactose will block cleavage. |
Contents : |
α(1-3,4)-Fucosidase in 20 mM Tris-HCl, 25 mM NaCl,(pH 7.5).
5× Reaction Buffer 5.0 (250 mM sodium phosphate, pH 5.0) |
Bio-activity : |
Activity: 0.5 U/mL
Specific Activity: > 2 U/mg |
Molecular Mass : |
40 kDa |
Suggested usage : |
1. Add up to 1 nmole of oligosaccharide to a tube.
2. Add de-ionized water to a total of 15 μL.
3. Add 4 μL of 5× Reaction Buffer 5.0.
4. Add 1 μL of α(1-3,4)-Fucosidase.
5. Incubate for 1 hour at 37 centigrade. |
Unit Definition : |
One unit of Fucosidase activity is defined as the amount of enzyme required to cleave 1 μmole of fucose from Lewis X trisaccharide, 4-methylumbelliferyl glycoside in 1 minute at 37 centigrade and pH 5.0. Lewis X trisaccharide is Gal β-(1-4)[Fuc α-(1-3)]GlcNAc. |
Purity : |
Each lot of α(1-3,4) Fucosidase is tested for contaminating activities by incubating the enzyme for 24 hours at 37 centigrade with the appropriate substrates; the detection limit of this assay is 5 μU/mL (IUB). A passing lot will have no detectable activity.
For the protease assay, 10 μg of denatured BSA is incubated for 24 hours with 2 μL of enzyme. Analysis of the BSA band after SDS-PAGE should show no evidence of degradation. |
Applications : |
α(1-3, 4) Fucosidase is useful for:
- Fucose linkage determination
- Deglycosylating glycoproteins with Lewis structures |
Stability : |
Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity. |
Storage : |
Store enzyme at 4 centigrade. Do not freeze. |
Storage Buffer : |
The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, 25 mM NaCl pH 7.5. |