Alpha-(1-6)-Fucosidase

Part number E-F006
Removes non-reducing, terminal branched fucose when linked alpha-(1-6) to GlcNAc of a GlcNAc-GlcNAc disaccharide structure that has been labeled with a fluorescent molecule. To date, the fluorescent labels ANTS, ANDSA and 2-AA have been demonstrated to function with the fucosidase. There is no cleavage of unlabeled oligosaccharides.

Source Elizabethkingia meningosepticum (was Chryseobacterium meningosepticum)
EC 3.2.1.51

Contents
60 µl aliquot of enzyme (30 mU) in 20 mM Tris-HCl, 25 mM NaCl,(pH 7.5).
200 µl 5x Reaction Buffer 5.0 (250 mM sodium phosphate, pH 5.0)

Specific Activity >1.5 U/mg
Activity 1.0 U/ml

Molecular weight 50,000 daltons

Suggested usage
1. Add up to 1 nmole of labeled 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 Alpha-(1-6)-Fucosidase.
5. Incubate for 1 hour at 37°C.

Specificity alpha(1-6) Fucosidase is specific for cleaving the alpha(1-6) core fucose from reducing terminally labeled oligosaccharides. To date, the fluorescent labels ANTS and ANDSA(aminonapthalene tri and disulfonic acid) and 2-AA(aminobenzoic acid) have been demonstrated to function with the fucosidase. There is no cleavage of unlabeled oligosaccharides whether or not they are free or glycopeptides. Oligosaccharides such as fucosylated trimannosylchitobiose can be cleave to completion using the recomended reaction conditions. However, longer oligosaccharides such as fucosylated biantennary oligosaccharides are only partially cleaved. The fucosidase will also cleave non-reducing terminal alpha 1-3 and alpha 1-4 fucose(but not alpha 1-2 fucose) provided the fucose is not branching. No labeling is required. These substrates do not apparently occur in nature.
Cleavage of labeled oligosaccharides by the fucosidase is diagnostic of alpha 1-6 core fucosylation.

Specific Activity One unit of Fucosidase activity is defined as the amount of enzyme required to cleave 1 µmole of fucose from 4−methylumbelliferyl-α-L-fucoside. in 1 minute at 37°C and pH 5.0.

Storage Store enzyme at 4°C.