Method for the measurement of endo-xylanase (XylX6 method)

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 ; 405

Signal response: increase 

Total assay time: 10 min

Detection limit: 5,3 x 10-4 U/ml

Reproducibility (%): ~ 3%

Stability: > 2 years under recommended storage conditions

The XylX6 assay reagent contains two components:
1) 4,6-O-(3-ketobutylidene)-4-nitrophenyl-β-D-45-glucosyl-xylopentaoside
2) β-xylosidase

The ketone blocking group prevents any hydrolytic action by the β-xylosidase or other exo-acting glycosidases on the XylX6 substrate. Incubation with an endo-xylanase generates a non-blocked colourimetric oligosaccharide that is rapidly hydrolysed by the ancillary β-Xxylosidase. The rate of formation of 4-nitrophenol is therefore directly related to the hydrolysis of XylX6 by the endo-xylanase. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH = 10,0).

Note that standard curves relating the absorbance obtained using the XylX6 assay to endo-xylanase activity on the native substrates, wheat arabinoxylan and beechwood xylan, are provided in the Supporting Information file under the Documentation tab.

Novel method
   

• Very cost effective
• All reagents stable for > 4 years
• Completely specific for endo-1,4-β-xylanase
• Generally applicable and highly sensitive
• Simple format
• Suitable for manual and auto-analyzer
• Excellent reproducibility
• Standard included

Enzyme extracts, fermentation broths or commercial enzyme preparations

Safety data sheet Validation report Data calculator Supporting information Data Booklet

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