产品资料

Megazyme/Cellulase Assay Kit (CellG3 Method)/K-CellG3/180 / 360 assays per kit / 720 (auto-analyser)

TheCellG3assayreagentforthemeasurementofendo-cellulase(endo-1,4-β-glucanase)containstwocomponents;
1)4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-D-cellotrioside(BCNPG3)and2)Thermostableβ-glucosidase.Thebenzylideneblockinggrouppreventsanyhydrolyticactionbytheβ-glucosidaseonBCNPG3. Incubationwithanendo-cellulasegeneratesanon-blockedcolourimetricoligosaccharidethatisrapidlyhydrolysedbytheancillary β-glucosidase. Therateofformationof2-chloro-4-nitrophenolisthereforedirectlyrelatedtothehydrolysisofBCNPG3bytheendo-cellulase. ThereactionisterminatedandthephenolatecolourisdevelopedonadditionofTrisbuffersolution(pH9.0).

Pleasenotethatanewassaykit(K-CellG5)isnowavailableforthemeasurementofendo-cellulase. TheCellG5reagentcontainsacellopentaosecoreandexhibitsvastlyimprovedsensitivityforsomecellulases. Inaddition,theexchangeofthebenzylideneblockinggroupinCellG3for3-keto-butylideneinCellG5improvesthesubstrate’swatersolubilitysignificantly,allowingforareductionintheconcentrationofDMSOrequiredintheassay. AsDMSOisknowntoinhibitcertaincellulases,thisisanotherbenefitinusingCellG5. MegazymenowrecommendstheuseofK-CellG5forallassaysforthemeasurementofendo-cellulase.

Suitableforauto/analyserformats.

Novelsubstratesforthemeasurementofendo-1,4-β-glucanase(endo-cellulase).

McCleary,B.V.,Mangan,D.,Daly,R.,Fort,S.,Ivory,R.&McCormack,N.(2014).CarbohydrateResearch,385,9-17.

LinktoArticle

ReadAbstract

Aspecificandsensitivesubstratefortheassayofendo-1,4-β-glucanase(cellulase)hasbeenprepared.Thesubstratemixturecomprisesbenzylideneend-blocked2-chloro-4-nitrophenyl-β-cellotrioside(BzCNPG3)inthepresenceofthermostableβ-glucosidase.Hydrolysisbyexo-actingenzymessuchasβ-glucosidaseandexo-β-glucanaseispreventedbythepresenceofthebenzylidenegrouponthenon-reducingendD-glucosylresidue.Onhydrolysisbycellulase,the2-chloro-4-nitrophenyl-β-glycosideisimmediatelyhydrolysedto2-chloro-4-nitrophenolandfreeD-glucosebytheβ-glucosidaseinthesubstratemixture.Thereactionisterminatedandcolourdevelopedbytheadditionofaweakalkalinesolution.Theassayprocedureissimpletouse,specific,accurate,robustandreADIlyadaptedtoautomation.Thisprocedureshouldfindwidespreadapplicationsinbiomassenzymologyandinthespecificassayofendo-1,4-β-glucanaseingeneral.

Quantitativefluorometricassayforthemeasurementofendo-1,4-β-glucanase.

Mangan,D.,McCleary,B.V.,Liadova,A.,Ivory,R.&McCormack,N.(2014).CarbohydrateResearch,395,47-51.

LinktoArticle

ReadAbstract

Thereisagrowingdemandforresearchtoolstoaidthescientificcommunityinthesearchforimprovedcellulaseenzymesforthebiofuelindustry.Inthiswork,wedescribeanovelfluorometricassayforcellulase(endo-1,4-β-glucanase)whichisbasedontheuseof4,6-O-benzylidene-4-methylumbelliferyl-β-cellotrioside(BzMUG3)inthepresenceofanancillaryβ-glucosidase.Thisassaycanbeusedquantitativelyoverareasonablelinearrange,orqualitativelyasasolutionscreeningtoolwhichmayfindextensiveuseintheareaofmetagenomics.

Anovelautomatableenzyme-coupledcolorimetricassayforendo-1,4-β-glucanase(cellulase).

Mangan,D.,Cornaggia,C.,McKie,V.,Kargelis.T.&V.McCleary,B.V.(2016).AnalyticalandBioanalyticalChemistry,408(15),4159-4168.

Linktoarticle

ReadAbstract

endo-1,4-β-Glucanase(endo-cellulase,EC3.2.1.4)isoneofthemostwidelyusedenzymesinindustry.Despiteitsimportance,improvedmethodsfortherapid,selective,quantitativeassayofthisenzymehavebeenslowtoemerge.In2014,anovelenzyme-coupledassaythataddressedmanyofthelimitationsoftheexistingassaymethodologywasreported.Thisinvolvedtheuseofabifunctionalsubstratechemicallyderivedfromcellotriose.Reportedhereinisamuchimprovedversionofthisassayemployinganovelsubstrate,namely4,6-O-(3-ketobutylidene)-4-nitrophenyl-β-D-cellopentaoside.

Colourimetricmethodforthedeterminationofendo-1,4-β-
glucanase(cellulase)inenzymepreparationsandfermentation
products

Principle:
                    (endo-1,4-β-glucanase)
(1)Benzylidene-G₃-β-CNP+H₂O→Blocked-G_X+G_(3-X)-β-CNP

        (thermostableβ-glucosidase)
(2)G_(3-X)-β-CNP+H₂O→D-glucose+CNP

   (alkalinesolution)
(3)CNP→phenolateion(yellowcolour)
Note:CNP=2-Chloro-4-nitrophenol

Kitsize:                          180/360assays/720(auto-analyser)
Method:                          Spectrophotometricat400nm
Totalassaytime:             ~20min
Detectionlimit:              0.05U/mL
Applicationexamples:
Fermentationbroths,industrialenzymepreparationsandbiofuelsresearch
Methodrecognition:     Novelmethod
Note:                                Theanalogousfluorimetricreagent(R-CELLFLR)
                                          isalsoavailablewith10-foldgreatersensitivity.

Advantages

Verycosteffective
Allreagentsstablefor>2yearsafterpreparation
Completelyspecificforcellulase(endo-1,4-glucanase).Thesubstrateisnothydrolysedbyβ-glucosidase,cellbiohyrolaseoranyotherenzymestested
KineticassayspossIBLeduetosignificantphenolateionpresence(andUVabsorbance)atpH5-6
Simpleformat.Wellsuitedtoautomation
Standardincluded

对照品标准品，生物试剂盒，gibco胎牛血清，试剂采购网，中国试剂网，细胞因子试剂盒

Megazyme/Cellulase Assay Kit (CellG3 Method)/K-CellG3/180 / 360 assays per kit / 720 (auto-analyser)

Novelsubstratesforthemeasurementofendo-1,4-β-glucanase(endo-cellulase).

Quantitativefluorometricassayforthemeasurementofendo-1,4-β-glucanase.

Anovelautomatableenzyme-coupledcolorimetricassayforendo-1,4-β-glucanase(cellulase).