| Species : |
Canavalia ensiformis |
| Source : |
Yeast |
| Description : |
α1-2,3,6 Mannosidase is a broad specificity exoglycosidase that catalyzes the hydrolysis of terminal, non-reducing α1-2, α1-3 and α1-6 linked mannose residues from oligosaccharides. |
| Molecular Mass : |
110 kDa |
| Purity : |
≥95% purity, as determined by SDS-PAGE and intact ESI-MS |
| Notes : |
1. α1-2,3,6 mannosidase is a glycosylated protein. The theoretical molecular weight is 110 kDa, and it contains two subunits of 66 kDa and 44 kDa, respectively.
2. The optimum pH value of α1-2,3,6 mannosidase is 4.5. Adding 2 mM Zn2+ into the reaction system can enhance the activity of the enzyme.
3. In order to completely remove all mannose residues linked by α at the non-reducing end, it may be necessary to increase the enzyme concentration and incubate overnight (18 hours).
4. In order to completely remove α1-6 mannose residues from complex substrates, it may be necessary to use α1-2,3,6 mannosidase and α1-6 mannosidase for enzyme digestion.
5. If a larger reaction volume is needed, the reaction system can be enlarged in proportion.
6. The amount of exonuclease is different with different substrates. In the reaction system of 10-25 μL, when 1 μg glycoprotein or 100 nM oligosaccharide is treated within 1 hour, the initial amount of enzyme ranges from 1-2 μL. If the enzyme digestion is not complete, overnight reaction is needed. |
| Storage : |
Optimal activity and stability for up to 24 months |
| Storage Buffer : |
20 mM Tris-HCl, 50 mM NaCl, pH 7.5 at 25 centigrade. |
| Unit definition : |
One unit refers to the amount of enzyme needed to hydrolyze more than 95% of terminal mannose in 1 nmol man (α 1,3)-man (β 1,4)-glcnac-7-amino-4-methyl-coumarin (AMC) in 10 μL total reaction system at 37 centigrade within one hour.
Detection condition of unit activity: In 10 μL reaction system, α1-2,3,6 mannosidase diluted twice was incubated with 1 nmol AMC labeled substrate in 1X GlycoBuffer 4 and 1× zinc. The reaction mixture was incubated at 37 centigrade for 1 hour. The separation of reaction products was observed by TLC. |
| Heat Inactivation : |
95 centigrade for 10 minutes |
| Reaction Buffer : |
1× GlycoBuffer 4, Supplement with 1× Zinc, Incubate at 37 centigrade.
1× GlycoBuffer 4: 50 mM sodium acetate (pH 4.5 at 25 centigrade) |
