Releases non-reducing terminal α(1-2,3,6)-linked mannose from oligosaccharides. The enzyme exhibits some kinetic preference for α(1-2,3)-linked mannose over α(1-6)-linked mannose.
The enzyme has broad specificity, cleaving α(1-2,3,6)-linked mannose, although some kinetic preference has been observed (α1-2, 6 > 3). The enzyme will not cleave a single α(1-6) linked mannose residue from core β-mannose, but will, however, remove a single α1-3 linked mannose from the core β-mannose. By using enzyme concentrations of around 50 U/ml and extended incubation times (up to 18 hours) at 37°C, complete removal of all non-reducing terminal α linked mannose residues may be achieved.
To expedite glycan sequencing studies, the sluggish activity of GKX-5010 Jack Bean α mannosidase toward α(1-6)-linked mannose residues can be overcome by using the enzyme in combination with the α mannosidase from Xanthomonas mannihotis which rapidly cleaves α(1-6) linkages. The X. mannihotis α-mannosidase may inhibit the action of other mannosidases if a branched (α1-6) mannose is present in the substrate, so is typically added after incubation of the substrate with Jack bean α-mannosidase (GKX-5010).
Figure 1: Cleavage specificity for GKX-5010 α(1-2,3,6)-Mannosidase.
WS0095 GKX-5010 Incubation Buffer (500 mM sodium acetate, 10 mM zinc chloride (pH 5.0).
5X concentrated buffer which when diluted gives 100 mM sodium acetate pH 5.0 containing 2 mM Zn2+.
20 mM Tris-HCl, 20 mM NaCl (pH 7.5)
MW: ~190 kD
One unit is defined as the amount of enzyme that will hydrolyze 1 μmole of pNP-α-mannopyranoside per minute at pH 4.5 and 37°C.
Size: 10 U (70 μl)
Concentration: ≥ 150 U/ml
Product Code: GKX-5010
Q. Can this enzyme cleave O-Linked mannose?
A. Yes, α(1-2,3,6)-Mannosidase from jack bean has been shown to be capable of digesting O-linked oligomannose chains on O-mannosylated glycoproteins, and can also cleave the Man-α-Ser/Thr glycosidic bond .
1. Gomathinayagam S, Hamilton SR. In vitro enzymatic treatment to remove O-linked mannose from intact glycoproteins. Appl Microbiol Biotechnol. 2014 Mar;98(6):2545-54. [PubMed]
Product Safety Documentation for GKX-5010:
|GKX-5010||Glyko® α(1-2,3,6)-Mannosidase (jack bean)|||
|WS0095||5X Reaction Buffer|
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