Reagent for the specific measurement of endo-cellulase (endo-1,4-β-glucanase)

Shipping temperature: ambient

Storage temperature:
- Short term stability: 2-8^oC,
- Long term stability: see individual component labels

Assay format: spectrophotometer, auto-analyser

Detection method: absorbance - 400

Signal response: increase

Total assay time: ~ 10 min

Detection limit: 1,2 x 10-3 U/ml

Reproducibility (%): ~ 3%

Stability: > 2 years under recommended storage conditions

The CellG5 assay reagent for the measurement of endo-cellulase (endo-1,4-β-glucanase) contains two components : 
1) 4,6-O-(3-ketobutylidene)-4-nitrophenyl-β-D-cellopentaoside (BPNPG5)
2) thermostable β-glucosidase.

The ketone blocking group prevents any hydrolytic action by the β-glucosidase on BPNPG5.  Incubation with an endo-cellulase generates a non-blocked colourimetric oligosaccharide that is rapidly hydrolysed by the ancillary β-glucosidase. The rate of formation of 4-nitrophenol is therefore directly related to the hydrolysis of BPNPG5 by the endo-cellulase. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH 9.0).
The CellG5 assay represents a huge step forward in the methodology for the measurement of cellulase that traditionally relied on substrates such as CM-cellulose, Avicel, cellooligosaccharides, filter paper or dyed polysaccharides including CMC Congo red or cellulose azure.

Novel method

• Very cost effective
• All reagents stable for > 4 years after preparation
• Completely specific for cellulase (endo-1,4-glucanase) 
• Generally applicable and highly sensitive 
• Simple format. Well suited to automation 
• Standard included

Fermentation broths, industrial enzyme preparations and biofuels research

Data Booklet Safety data sheet Validation report Data calculator

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