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Megazyme/Cellulase Assay Kit (CellG5 Method)/K-CellG5-4V/120 / 240 assays (manual) / 480 (auto-analyser)
TheCellG5assayreagentforthemeasurementofendo-cellulase(endo-1,4-β-glucanase)containstwocomponents;
1)4,6-O-(3-Ketobutylidene)-4-nitrophenyl-β-D-cellopentaoside(BPNPG5)and2)Thermostableβ-glucosidase.Theketoneblockinggrouppreventsanyhydrolyticactionbytheβ-glucosidaseonBPNPG5. Incubationwithanendo-cellulasegeneratesanon-blockedcolourimetricoligosaccharidethatisrapidlyhydrolysedbytheancillaryβ-glucosidase.Therateofformationof4-nitrophenolisthereforedirectlyrelatedtothehydrolysisofBPNPG5bytheendo-cellulase.ThereactionisterminatedandthephenolatecolourisdevelopedonadditionofTrisbuffersolution(pH9.0).
TheCellG5assayrepresentsahugestepforwardinthemethodologyforthemeasurementofcellulasethattrADItionallyreliedonsubstratessuchasCM-cellulose,Avicel,cellooligosaccharides,filterpaperordyedpolysaccharidesincludingCMCCongoredorcelluloseazure.
Novelsubstratesforthemeasurementofendo-1,4-β-glucanase(endo-cellulase).
Quantitativefluorometricassayforthemeasurementofendo-1,4-β-glucanase.
Anovelautomatableenzyme-coupledcolorimetricassayforendo-1,4-β-glucanase(cellulase).
CompletegenomesequenceofBacillussp.275,producingextracellularcellulolytic,xylanolyticandligninolyticenzymes.
Colourimetricmethodforthedeterminationof
endo-1,4-β-glucanase(cellulase)inenzymepreparationsandfermentationproducts
Principle:
(endo-1,4-β-glucanase)
(1)3-Ketobutylidene-G5-β-PNP+H2O→Blocked-GX +G(5-X)-β-PNP
(thermostableβ-glucosidase)
(2)G(5-X)-β-PNP+H2O→D-glucose+PNP
(alkalinesolution)
(3)PNP→phenolateion(yellowcolour)
Note: PNP=4-nitrophenol
Kitsize:
K-CellG5-4V120/240assays(manual)/480(auto-analyser)
or
K-CellG5-2V60/120assays(manual)/240(auto-analyser)Method: Spectrophotometricat400nm
Totalassaytime: 10min
Detectionlimit: 3.5x10-4U/mL
Applicationexamples:
Fermentationbroths,industrialenzymepreparationsandbiofuelsresearch
Methodrecognition: Novelmethod
Advantages
- Verycosteffective
- Allreagentsstablefor>4years
- Completelyspecificforcellulase(endo-1,4-glucanase)
- Generallyapplicableandhighlysensitive
- Simpleformat.Wellsuitedtoautomation
- Standardincluded