PNGaseF,peptide-N-glycosidaseF,peptide-N4-(N-acetyl-β-glucosaminyl)asparagineamidase. ReleasesintactN-glycansbycleaving betweentheinnermostGlcNAcandAsn. ThesameformulationprovidedinGlykoPrep™, ≥10U/mlandEDTA-free.
Source:
RecombinantgenefromElizabethkingiameningosepticum,expressedinE.coli.Thesourceorganismwaspreviouslyknownas Chryseobacterium[Flavobacterium]meningosepticum[1].
Specificity:
CleavesallN-linkedcomplex,hybridorhighmannoseoligosaccharides,unlessα(1-3)corefucosylated,asinplantglycans.Asparaginemustbepeptide-bondedatbothtermini.Phosphate,sulfateandsialicacidgroupsattachedtotheoligosaccharidedonotaffectcleavage.EndoFfree.Highlyconcentrated,particularlyusefulfordeglycosylationundernativeconditions.
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Figure1:CleavagespecificityforGKE-5020N-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.
"B"packsizeshipswith:
Formulation:
20mMTrisHClpH7.5,50mMNaCl
pHoptimum:
8.6
pHrange:
7.5-9.5
UnitDefinition:
OneunitisdefinedastheamountofenzymerequiredtocatalyzethereleaseofN-linkedoligosaccharidesfrom1μmoleofdenaturedribonucleaseBperminuteat37°C,pH7.5.
Size:400mUin40µl
Concentration: ≥10U/ml
ProductCode:GKE-5020B
NotformulatedforFACE®Analysis,useGK50052.
1.KimKK,KimMK,LimJH,ParkHY,LeeST. TransferofChryseobacteriummeningosepticumandChryseobacteriummiricolatoElizabethkingiagen.nov.asElizabethkingiameningosepticacomb.nov.andElizabethkingiamiricolacomb.nov. IntJSystEvolMicrobiol.2005May;55(Pt3):1287-93.
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