Description

PNGaseF,peptide-N-glycosidaseF,peptide-N⁴-(N-acetyl-β-glucosaminyl)asparagineamidase. ReleasesintactN-glycansbycleaving betweentheinnermostGlcNAcandAsn.

Source:
RecombinantgenefromElizabethkingiameningosepticum,expressedinE.coli.Thesourceorganismwaspreviouslyknownas Chryseobacterium Chryseobacterium[Flavobacterium]meningosepticum[1].

Specificity: 
CleavesallN-linkedcomplex,hybridorhighmannoseoligosaccharides,unlessα(1-3)corefucosylated,asinplantglycans.Asparaginemustbepeptide-bondedatbothtermini.Phosphate,sulfateandsialicacidgroupsattachedtotheoligosaccharidedonotaffectcleavage.EndoFfree.Highlyconcentrated,particularlyusefulfordeglycosylationundernativeconditions. 

Figure1:CleavagespecificityforGKE-5010N-Glycanase(PNGaseF).
R₁:N&CsubstitutionotherthanH
R₂: maybeHortherestofanoligosaccharide
R₃: maybeHor α(1-6)-linkedfucose

Applications: 
Toobtainefficientdeglycosylationofglycoproteinsubstratesundernon-denaturingconditions,itisnecessary touseahigherstartingconcentrationofenzyme.N-Glycanase®PLUSandULTRA(EDTA-Free)aresuppliedat≥10U/ml,andarerecommendedforallapplicationsrequiringdeglycosylationofglycoproteinsintheabsenceofdenaturants.Thehighactivityalsoallowssmaller reactionvolumesandshorterreactiontimestobeexplored.

Shipswith: 

• 1eaWS00105xN-GlycanaseReactionBuffer:100mMsodiumphosphatepH7.5,0.1%sodiumazide
• 1eaWS0012DenaturationSolution: 2%SDS,1M2-mercaptoethanol
• 1eaWS0013DetergentSolution:15%Detergent
• 1eaWS01455xN-GlycanaseTrisReactionBuffer:50mMTris-HClpH8.0

Alsoavailablein"D"(1U)packsize,whichdoesnotshipwithbuffers.

Formulation:
20mMTrisHClpH7.5,containing1mMEDTAand50mMNaCl

pHoptimum:
8.6

pHrange:
7.5-9.5

UnitDefinition:
OneunitisdefinedastheamountofenzymerequiredtocatalyzethereleaseofN-linkedoligosaccharidesfrom1 μmoleofdenaturedribonucleaseBperminute atpH7.5and37°C.

Size:400mU(40µl)

Concentration: ≥10U/ml

ProductCode:GKE-5010B

NotformulatedforFACE®Analysis,useGK50052.

1.KimKK,KimMK,LimJH,ParkHY,LeeST. TransferofChryseobacteriummeningosepticumandChryseobacteriummiricolatoElizabethkingiagen.nov.asElizabethkingiameningosepticacomb.nov.andElizabethkingiamiricolacomb.nov. IntJSystEvolMicrobiol.2005May;55(Pt3):1287-93.