內(nèi)切纖維素酶檢測(cè)試劑盒[CELLG3方法] Cellulase Assay Kit (CELLG3 Method) 貨號(hào)：K-CELLG3 Megazyme中文站

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內(nèi)切纖維素酶檢測(cè)試劑盒[CELLG3方法]

英文名：Cellulase Assay Kit (CELLG3 Method)

貨號(hào)：K-CELLG3

規(guī)格：180 / 360 assays per kit / 720 (auto-analyser)

 纖維素酶是一種重要的酶產(chǎn)品，是一種復(fù)合酶，主要由外切β-葡聚糖酶、內(nèi)切β-葡聚糖酶和β-葡萄糖苷酶等組成，還有很高活力的木聚糖酶。

提供內(nèi)切纖維素酶檢測(cè)試劑盒，色度法（400 nm）測(cè)定酶制品和發(fā)酵產(chǎn)品中的纖維素酶（內(nèi)切-1,4-β-葡聚糖酶）。

內(nèi)切纖維素酶檢測(cè)試劑盒(CELLG3方法) K-CELLG3

使用高純度β-葡萄糖苷酶和苯亞甲基阻斷，2 – 氯-4 – 硝基苯基-β-Dcellotrioside（BClPNPβ-G3）。 β-葡萄糖苷酶的能力確保測(cè)定的可靠的最大的靈敏度。BClPNPβ-G3的水解為苯亞甲基，通過(guò)纖維素酶阻斷纖維二糖和2-氯-4 -硝基苯基-β-D-葡萄糖， 2-Cl-4-硝基苯基-β-D-葡萄糖通過(guò)β-葡萄糖苷酶立即裂解為D-葡萄糖和游離的2 – 氯-4 – 硝基苯酚（ClPNP）。

反應(yīng)時(shí)間：16min

適用樣品：酶制品和發(fā)酵產(chǎn)品

優(yōu)點(diǎn)：

價(jià)格低廉（每次檢測(cè)成本）

特異性高

方法簡(jiǎn)單

包含標(biāo)準(zhǔn)品

 

 The CELLG3 assay reagent for the measurement of endo-cellulase (endo-1,4-β-glucanase) contains two components; 

1) 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-D-cellotrioside (BCNPG3) and 2) thermostable β-glucosidase. The benzylidene blocking group prevents any hydrolytic action by the β-glucosidase on BCNPG3.  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 2-chloro-4-nitrophenol is therefore directly related to the hydrolysis of BCNPG3 by the endo-cellulase.  The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH 9.0).
Please note that a new assay kit (K-CELLG5) is now available for the measurement of endo-cellulase.  The CELLG5 reagent contains a cellopentaose core and exhibits vastly improved sensitivity for some cellulases.  In addition, the exchange of the benzylidene blocking group in CELLG3 for 3-keto-butylidene in CELLG5 improves the substrate’s water solubility significantly, allowing for a reduction in the concentration of DMSO required in the assay.  As DMSO is known to inhibit certain cellulases, this is another benefit in using CELLG5.  Megazyme now recommends the use of K-CELLG5 for all assays for the measurement of endo-cellulase.

Colourimetric method for the determination of 
endo-1,4-β-glucanase (cellulase) in enzyme preparations and fermentationproducts

Principle:
                                       (endo-1,4-β-glucanase)
(1) 3-Ketobutylidene-G5-β-PNP + H2O → Blocked-GX + 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

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