内切纤维素酶检测试剂盒[CELLG5方法]

英文名：Cellulase Assay Kit (CELLG5 Method)

货号：K-CELLG5-2V

规格：60 / 120 assays (manual) / 240 (auto-analyser)

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) and 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).

Colourimetric method for the determination of

endo-1,4-β-glucanase (cellulase) in enzyme preparations and fermentation products

Principle:

(endo-1,4-β-glucanase)

(1) 3-Ketobutylidene-G₅-β-PNP + H₂O → Blocked-G_X + G_(5-X)-β-PNP

(thermostable β-glucosidase)

(2) G_(5-X)-β-PNP + H₂O → D-glucose + PNP

(alkaline solution)

(3) PNP → phenolate ion (yellow colour)

Note: PNP = 4-nitrophenol

Kit size:

K-CELLG5-4V 120 / 240 assays (manual) / 480 (auto-analyser)

or

K-CELLG5-2V 60 / 120 assays (manual) / 240 (auto-analyser)

Method: Spectrophotometric at 400 nm

Total assay time: 10 min

Detection limit: 3.5 x 10^-4 U/mL

Application examples:

Fermentation broths, industrial enzyme preparations, biofuels research

Method recognition: Novel method

Advantages

• Very cost effective

• All reagents stable for > 4 years

• Completely specific for cellulase (endo-1,4-glucanase)

• Generally applicable and highly sensitive

• Simple format. Well suited to automation

• Standard included

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