α-(1,6) core mannosidase
α(1-6) Core Mannosidase cleaves unbranched non-reducing terminal mannose, α(1-6) linked to the β-linked core mannose of the conserved mannosylchitobiose core of N-linked oligosaccharides. The presence of fucose linked to the core N-acetylglucosamine has no effect on cleavage.
The enzyme may inhibit other mannosidases, such as α(1-2,3,6) mannosidase (E-AM01), if a noncleavable α(1-6) mannose is present on the substrate. It should therefore always be added subsequent to digestion by other mannosidases.
α(1-6) Core Mannosidase is useful for determining mannose linkages and rapidly removing the α(1-6) core mannose resistant to other mannosidases
α(1-6) Core Mannosidase is useful for:
Determining mannose linkages
Rapidly removing the α(1-6) core mannose resistant to other mannosidases
Source: recombinant from Xanthamonas manihotis E. Coli
Alternate Names: α-D-mannosidase, p-nitrophenyl-α-mannosidase, α-D-mannopyranosidase, core α mannosidase, exo-α-mannosidase
Core Alpha Mannosidase in 50 mM sodium phosphate, 0.1 mM ZnCl (pH 7.5).
200 µL 5x Reaction Buffer 5.0 (250 mM sodium phosphate, pH 5.0).
The enzyme is provided as a sterile solution in 50 mM sodium phosphate pH 7.5, 0.1 mM ZnCl2.
Specific Activity: >1.5 U/mg
Activity: 1 U/mL
Molecular weight: ~52,000 daltons
1. Add up to 1 nmol of oligosaccharide.
2. Add deionized water to 15 µL.
3. Add 4 µL 5x Reaction Buffer 5.0.
4. Add 1 µL of enzyme
5. Incubate ten minutes at 37°C.
Specific Activity Assay: One unit of α(1-6) Mannosidase is defined as the amount of enzyme required to produce 2 µmoles of mannose in 1 minute at 37°C, pH 5.0 from α(1-6) mannobiose.
Storage: Store enzyme at 4°C.
Purity: Each lot of α(1-6) mannosidase is tested for contaminating substances by incubating the enzyme for 24 hours at 37°C with substrates indicated in the table below. No detectable activity is evident for any of these potential contaminants. The detection limit of this assay is 5 µU/mL (IUB).
The production host strain has been extensively tested and does not produce any detectable glycosidases.
Stability: Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.