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For the specific measurement of cellulase (endo-1,4-β-glucanase) in enzyme preparations and fermentation products.
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) Benzylidene-G3-β-CNP + H2O → Blocked-GX + G(3-X)-β-CNP
(thermostable β-glucosidase)
(2) G(3-X)-β-CNP + H2O → D-glucose + CNP
(alkaline solution)
(3) CNP → phenolate ion (yellow colour)
Note: CNP = 2-Chloro-4-nitrophenol
Kit size: 180 / 360 assays / 720 (auto-analyser)
Method: Spectrophotometric at 400 nm
Total assay time: ~ 20 min
Detection lim-it: 0.05 U/mL
Application examples:
Fermentation broths, industrial enzyme preparations, biofuels research
Method recognition: Novel method
Note: The analogous fluorimetric reagent (R-CELLF)
is also available with 10-fold greater sensitivity
Advantages
- Very cost effective
- All reagents stable for > 2 years after preparation
- Completely specific for cellulase (endo-1,4-glucanase). The substrate is not hydrolysed by β-glucosidase, cellbiohyrolase or any other enzymes tested
- Kinetic assays possible due to significant phenolate ion presence (and UV absorbance) at pH 5-6
- Simple format. Well suited to automation
- Standard included
爱尔兰Megazyme公司是全球知名的酶及酶法分析试剂盒的生产厂家，其中大多数产品都被AOAC(Association of Official Analytical Chemists)、AACC(American Association of Cereal Chemists)、ICC(International Association for Cereal Science and Technology)、 EBC(European Brewing Convention)、RACI(Royal Australian Chemical Institute)等国际组织采用。其中欧洲检测果汁及葡萄酒等发酵产品基本都采用该公司产品。 MEGAZYME公司产品分以下系列： -------------------------------------------------------------------------------- 1. 生物及食品酶法分析试剂盒 (Diagnostic Kits) 2. 试剂混合物(Reagent Mixtures) 3. 协同因子(Cofactors) 4. 糖酶片剂试验(Carbohydrase Tablet Tests) 5. 蛋白酶片剂试验(Protease Test Tablets) 6. 可溶性呈色底物(Soluble Chromogenic Substrates) 7. 不可溶性（交联的）呈色底物(Insoluble (Crosslinked) Chromogenic Substrates) 8. 酶(Enzymes) 9. 植物凝血素(Lectins) 10. 多聚糖(Polysaccharides) 11. 低聚糖(Oligosaccharides) 12. 相关产品 (General) -------------------------------------------------------------------------------- 其中产品被国际机构采用，建立国际检测方法如下： -------------------------------------------------------------------------------- 总淀粉检测盒（Total Starch Assay Kit） • AOAC Method 996.11 • AACC Method 76-13 • ICC Standard Method no. 168 • RACI Standard Method 总膳食纤维检测盒（Total Dietary Fibre Assay Kit） • AOAC Method 985.29 • AOAC Method 991.42 • AOAC Method 993.19 • AOAC Method 991.43 • AACC Method 32.07 • AACC Method 32.21 • AACC Method 32.05 淀粉损失检测盒（Starch Damage Assay Kit） • AACC Method 76-31 • ICC Standard No. 164 • RACI Standard Method α-淀粉酶检测盒Ceralpha (α-Amylase) Assay Kit • AOAC Method 2002.01 • ICC Standard No. 303 • RACI Standard Method • CCFRA Flour Testing Working Group Method 0018 • AACC - under evaluation. Alpha淀粉酶检测盒Amylazyme (α-Amylase Assay) • AACC Method 22.05 • RACI Standard Method 果聚糖检测盒Fructan Assay Kit • AOAC Method 999.03 • AACC Method 32-32 木聚糖酶AX片剂（Xylazyme AX Tablets） • Xylazyme AX Tablets have been widely adopted in the fermentation and feeds industries for the measurement of Xylanase. β-葡聚糖检测盒β-Glucan (Mixed linkage) Assay Kit • AACC Method 32-23 • EBC Methods 3.11.1, 4.16.1 and 8.11.1 • AOAC Method 995.16 • ICC Standard No. 166 • RACI Standard Method 极限糊精酶和支链淀粉酶检测片剂（Limit-Dextrizyme） • RACI Standard Method β-葡聚糖酶检测盒（β-Glucanase (Malt and Microbial) Assay Kit） • RACI Standard Method β-淀粉酶检测盒（Betamyl (β-Amylase Assay Kit)） • RACI Standard Method. 木聚糖酶检测底物（Azo-Wheat Arabinoxylan ） • Azo-Wheat Arabinoxylan, Azo-CM-Cellulose and Ceralpha methods for the assay of Xylanase, cellulase and ?-Amylase, respectively, have been adopted by the UK silage industry. 抗性淀粉检测盒（Resistant Starch Assay Kit） • AOAC Method 2002.02