The XylX6 assay reagent for the measurement of endo-xylanase (endo-1,4-β-xylanase) contains two components; 1) 4,6-O-(3-Ketobutylidene)-4-nitrophenyl-β-D-4⁵-glucosyl-xylopentaoside and 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-xylosidase. 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).

Colourimetric method for the determination of endo-1,4-β-xylanase in any 
sample type including crude enzyme extracts, fermentation broths or 
commercial enzyme preparations

Principle:
                                               (endo-1,4-β-xylanase)
(1) 3-Ketobutylidene-GX₅-β-PNP + H₂O → Blocked-GX_Y + X_(5-Y)-β-PNP

                            (β-glucosidase)
(2) X_(5-Y)-β-PNP + H₂O → D-xylose + PNP

       (alkaline solution)
(3) PNP → phenolate ion (yellow colour)
Note: PNP = 4-nitrophenol

Kit size:
K-XylX6-1V                        100 assays (manual) / 200 (auto-analyser)
                                           or
K-XylX6-2V                        200 assays (manual) / 400 (auto-analyser)
Method:                            Spectrophotometric at 400/405 nm
Total assay time:             10 min
Detection limit:                 5.3 x 10-4 U/mL
Application examples: 
Fermentation broths, industrial enzyme preparations, animal feed, biofuels research, barley 
malt analysis
Method recognition:        Novel method

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