Megazyme/1,2αLFucosidase (microbial) /EFUCM/3 Units at 37^oC

Highpurityrecombinant 1,2-α-L-Fucosidase(microbial)foruseinresearch,biochemicalenzymeassaysandinvitrodiagnosticanalysis.

EC3.2.1.63
CAZyFamily:GH95
CAS:37288-45-2

1,2-alpha-L-fucosidase;2-alpha-L-fucopyranosyl-beta-D-galactosidefucohydrolase

Recombinant.Fromamicrobialsource.
Insolution(Tris.HCl/NaCl/EDTA).
Suppliedat~12U/mL.

Specificactivity:
~160U/mg(37^oC,pH6.5on2’-fucosyllactose)

Stability: > 3yearsat4^oC.

ComparisonsofCaenorhabditisfucosyltransferasemutantsrevealamultiplicityofisomericN-glycanstructures.

Yan,S.,Jin,C.,Wilson,I.B.&Paschinger,K.(2015).JournalofProteomeResearch,14(12),5291-5305.

LinktoArticle

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RecentstudieshaveshownaremarkabledegreeofplasticityintheN-glycomeofthemodelnematodeCaenorhabditiselegans;ablationofglycosylation-relevantgenescanresultinradicallyalteredN-glycanprofilesdespiteonlyminorbiologicalphenotypiceffects.UptofourfucoseresiduesandfivedifferentlinkagesoffucoseareknownontheN-glycansofC.elegans.Duetothecomplexityinthewildtype,weestablishedthreemutantstrainsdefectiveintwocorefucosyltransferaseseach(fut-1;fut-6,fut-1;fut-8,andfut-6;fut-8).EnzymaticallyreleasedN-glycansweresubjecttoHPLCandMALDI-TOFMS/MS,incombinationwithvarioustreatments,toverifystructuraldetails.TheN-glycomeofthefut-1;fut-6mutantwasthemostcomplexofthethreedouble-mutantstrainsduetotheextensionofthecoreα1,6-fucoseaswellasthepresenceoffucoseonthebisectinggalactose.Incontrast,maximallytwofucoseswerefoundonN-glycansofthefut-1;fut-8andfut-6;fut-8strains.Thedifferentlocationsandcappingoffucosemeantthatupto13isomericstructures,manyhighlygalactosylated,weredeterminedforsomesinglemasses.ThesedatanotonlyshowthehighvariabilityoftheN-glycomiccapacityofa“simple”nematodebutalsoexemplifytheneedformultipleapproachestorevealindividualglycanstructureswithincomplexinvertebrateglycomes.

AnalysisofInvertebrateandProtistN-Glycans.

Hykollari,A.,Paschinger,K.,Eckmair,B.&Wilson,I.B.(2017).High-ThroughputGlycomicsandGlycoproteomics:MethodsandProtocols,1503,167-184.

LinktoArticle

ReadAbstract

N-glycansfrominvertebratesandprotistshaveoftenunusualstructureswhichpresentanalyticalchallenges.Bothcoreandantennalmodificationscanbequitedifferentfromthemorefamiliarvertebrateglycanmotifs;thereby,contrarytotheconceptthat“simple”organismshave“simple”N-glycans,rathercomplexoligosaccharidesstructures,includingzwitterionicandanionicones,havebeenfoundinarangeofspecies.Thus,tofacilitatetheoptimizedelucidationofthemaximalpossiblerangeofstructures,theanalyticalworkflowforglycomicsoftheseorganismsshouldincludesequentialreleaseandfractionationsteps.Peptide:N-glycosidaseFissufficienttoisolateN-glycansfromfungiandsomeprotists,butinmostinvertebratescoreα1,3-fucoseispresent,soreleaseoftheglycansfromglycopeptideswithpeptide:N-glycosidasesAisrequired.Subsequentsolid-phaseextractionwithgraphitizedcarbonandreversedphaseresinsenablesdifferentclassesofN-glycanstobeseparatedpriortohigh-pressureliquidchromatography(HPLC)andmatrix-assistedlaserdesorption/ionizationtime-of-flightmassspectrometry(MALDI-TOFMS).Dependingonthetypesandnumbersofglycanspresent,eitherreversed-ornormal-phaseHPLC(orbothinseries)enableevensingleisomericorisobaricstructurestobeseparatedpriortoMALDI-TOFMSandMS/MS.Theuseofenzymaticorchemicaltreatmentsallowsfurtherinsightstobegained,althoughsomeglycanmodifications(especiallymethylation)areresistant.Usingabatteryofmethods,sometimesupto100structuresfromasingleorganismcanbeassigned,acomplexitywhichraisesevolutionaryquestionsregardingthefunctionoftheseglycans.

DESCRIPTION

1,2-α-L-Fucosidase(microbial)

EC3.2.1.63
CAZyFamily:GH95
CAS: 37288-45-2

Synonyms:
1,2-alpha-L-fucosidase;2-alpha-L-fucopyranosyl-beta-D-galactosidefucohydrolase

Form:
Insolution(Tris.HCl/NaCl/EDTA).

Stability:
>3yearsat4^oC.

Specificactivity:
~160U/mg(37^oC,pH6.5on2’-fucosyllactose).

Unitdefinition:
OneUnitofα-1,2-L-fucosidaseactivityisdefinedastheamountofenzymerequiredtoreleaseoneμmolofα-L-fucoseperminutefrom2’-fucosyllactose(2mM)insodiumphosphatebuffer(100mM),pH6.5at37^oC.

Specificity:
Highlyspecificfornon-reducingterminalL-fucoseresidueslinkedtoD-galactoseresiduesbya1,2-α-linkage.Doesnothydrolysep-nitrophenyl-α-L-fucopyranoside.

Applications:
Foruseinglycobiologyresearch.

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Megazyme/1,2αLFucosidase (microbial) /EFUCM/3 Units at 37oC

ComparisonsofCaenorhabditisfucosyltransferasemutantsrevealamultiplicityofisomericN-glycanstructures.

AnalysisofInvertebrateandProtistN-Glycans.

Megazyme/1,2αLFucosidase (microbial) /EFUCM/3 Units at 37^oC