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.
α(1-3, 4) Fucosidase is useful for:
- Fucose linkage determination
- Deglycosylating glycoproteins with Lewis structures
Source: Isolated from Xanthamonas manihotis
Alternate Names: α-L-fucoside fucohydrolase, α-L-fucosidase, α-(1-3,4) fucosidase
α(1-3,4)-Fucosidase in 20 mM Tris-HCl, 25 mM NaCl,(pH 7.5).
5x Reaction Buffer 5.0 (250 mM sodium phosphate, pH 5.0)
Specific Activity: >2 U/mg
Activity: 0.5 U/mL
Molecular weight: 40,000 daltons
Formulation: The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, 25 mM NaCl pH 7.5.
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 5x Reaction Buffer 5.0.
4. Add 1 µL of α(1-3,4)-Fucosidase.
5. Incubate for 1 hour at 37°C.
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.
Specific Activity Assay: 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°C and pH 5.0. Lewis X trisaccharide is Gal β-(1-4)[Fuc α-(1-3)]GlcNAc.
Storage: Store enzyme at 4°C.
Purity: Each lot of α(1-3,4) Fucosidase is tested for contaminating activities by incubating the enzyme for 24 hours at 37°C 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.
Stability: Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.