Description:
α1-2,3,6Mannosidase,clonedfromJackBean,andalsoknownasJBM,isabroadspecificityexoglycosidasethatcatalyzesthehydrolysisofterminalα1-2,α1-3andα1-6linkedmannoseresiduesfromoligosaccharides.α1-2,3,6Mannosidasehasaslightpreferenceforα1-2mannoseresiduesoverα1-3andα1-6mannoseresidues.
ProductSource
ClonedfromCanavaliaensiformis(JackBean)andexpressedinPichiapastoris(1).ReagentsSupplied
Thefollowingreagentsaresuppliedwiththisproduct:
| Storeat(°C) | Concentration |
GlycoBuffer4 | -20 | 10X |
Zinc | -20 | 10X |
Notes:
α1-2,3,6Mannosidaseisaglycosylatedprotein.Thetheoreticalmolecularweightis110kDa,withtwosubunitsat66kDaand44kDa(3).TheoptimalpHforα1-2,3,6Mannosidaseis4.5.Enzymeactivityisenhancedbythepresenceof2mMZn2+inthereaction.Inordertoachievecompleteremovalofallnon-reducingterminalα-linkedmannoseresidues,itmaybenecessarytouseanincreasedenzymeconcentrationaswellasanovernightincubationtime(18hours).Toachievecompleteremovalofα1-6mannoseresiduesfromacomplexsubstrate,asequentialdigestusingα1-2,3,6Mannosidasefollowedbyα1-6Mannosidase(NEB#P0727)maybenecessary. Reactionsmaybescaled-uplinearlytoaccommodatelargerreactionvolumes.Theamountofexoglycosidaseenzymerequiredvarieswhendifferentsubstratesareused.Startwith1-2μlfor1μgofglycoproteinor100nMofoligosaccharideforonehourina10-25μlreaction.Ifthereisstillundigestedmaterial,letthereactiongoovernight.