alpha-L-fucosidefucohydrolase,alpha-L-fucosidase,alpha-(1-3,4)fucosidase

Alpha(1-3,4)Fucosidasecleavesbranchednon-reducingterminalfucose,linkedα(1-3)orα(1-4)totheN-acetylglucosamineofterminalGal-GlcNAcdisaccharidestructures.Thepresenceofsialicacid(butnotfucose)linkedtothegalactosewillblockcleavage.

Forremovingcorefucoselinkedα-(1-6)tothecoreGlcNAcofaGlcNAc-GlcNAcdisaccharidestructurewerecommendourAlpha-(1-6)Fucosidase(E-F006).

α(1-3,4)Fucosidaseisusefulfor:
nbsp;nbsp;Fucoselinkagedetermination
nbsp;nbsp;DeglycosylatingglycoproteinswithLewisstructures

SourceIsolatedfromXanthamonasmanihotis

EC3.2.1.51

Contents
Alpha-(1-3,4)-Fucosidasein20mMTris-HCl,25mMNaCl,(pH7.5).

Includedwith20µLand60µLpacksizes:
5xReactionBuffer5.0(250mMsodiumphosphate,pH5.0)

SpecificActivity>2U/mg
Activity0.5U/ml

Molecularweight40,000daltons

FormulationTheenzymeisprovidedasasterile-filteredsolutionin20mMTris-HCl,25mMNaClpH7.5.

Suggestedusage
1.Addupto1nmoleofoligosaccharidetoatube.
2.Addde-ionizedwatertoatotalof15µl.
3.Add4µlof5xReactionBuffer5.0.
4.Add1µlofAlpha-(1-3,4)-Fucosidase.
5.Incubatefor1hourat37˚C.

SpecifictityNon-reducingterminalbranchedfucosewhenlinkedalpha-(1-3)oralpha-(1-4)toGlcNAcofaGal-GlcNAcdisaccharidestructure.Thepresenceofsialicacid(butnotfucose)linkedtothegalactosewillblockcleavage.

SpecificActivityAssayOneunitofFucosidaseactivityisdefinedastheamountofenzymerequiredtocleave1µmoleoffucosefromLewisXtrisaccharide,4-methylumbelliferylglycosidein1minuteat37˚CandpH5.0.LewisXtrisaccharideisGalBeta-(1-4)[Fucalpha-(1-3)]GlcNAc.

StorageStoreenzymeat4˚C.

PurityEachlotofα(1-3,4)Fucosidaseistestedforcontaminatingactivitiesbyincubating
theenzymefor24hoursat37°Cwiththeappropriatesubstrates;thedetectionlimitofthisassayis5μU/mL(IUB).Apassinglotwillhavenodetectableactivity.

Fortheproteaseassay,10μgofdenaturedBSAisincubatedfor24hourswith2μLofenzyme.AnalysisoftheBSAbandafterSDS-PAGEshouldshownoevidenceofdegradation.

StABIlityStableatleast12monthswhenstoredproperly.Severaldaysexposuretoambienttemperatureswillnotreduceactivity.

Alpha-(1-3,4)FucosidaseReferences

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