内切纤维素酶检测试剂盒(CELLG5方法)

用于测量内切纤维素酶（内切-1,4-β-葡聚糖酶）的CellG5测定试剂包含两种成分：1）4,6-O-（3-酮丁叉基）-4-硝基苯基-β-D-纤维五糖苷（BPNPG5）和2）热稳定性β-葡萄糖苷酶。酮基阻断基团可防止β-葡萄糖苷酶对BPNPG5的任何水解作用。与内切纤维素酶一起孵育会产生一种未被阻断的比色寡糖，该寡糖可被辅助β-葡萄糖苷酶迅速水解。因此，4-硝基苯酚的形成速率与内切纤维素酶对BPNPG5的水解直接相关。反应终止后，加入Tris缓冲液（pH 9.0）即可使酚盐显色。

 

CellG5测定法在纤维素酶测定方法上迈出了巨大的一步，传统方法依赖于CM-纤维素、微晶纤维素、纤维寡糖、滤纸或染色多糖（包括CMC刚果红或纤维素天蓝）等底物。

优势

非常划算

所有试剂的稳定期均大于4年

对纤维素酶（内切-1,4-葡聚糖酶）具有完全特异性

普遍适用且高度敏感

格式简洁。非常适合自动化

已包含标准配置

内容：（K-CellG5-4V）120次/240次手动测定/480次自动分析仪测定

运输温度：常温

储存温度：短期稳定性：2-8℃，

长期稳定性：请参阅各组件标签

稳定性：在推荐的储存条件下，可保持>2年

分析物：内切纤维素酶

测定格式：分光光度计，自动分析仪

检测方法：吸光度

波长（纳米）：400

信号响应：增加

检测限：1.2 x 10-3 U/mL

再现性（%）：约3%

总测定时间：10分钟

应用示例：发酵液、工业酶制剂和生物燃料研究。

方法识别：新颖方法

 

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).

 

The CellG5 assay represents a huge step forward in the methodology for the measurement of cellulase that traditionally relied on substrates such as CM-cellulose, Avicel, cellooligosaccharides, filter paper or dyed polysaccharides including CMC Congo red or cellulose azure.

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

Content: (K-CellG5-4V) 120 / 240 assays (manual) / 480 assays (auto-analyser) 

Shipping Temperature: Ambient

Storage Temperature: Short term stability: 2-8oC,

Long term stability: See individual component labels

Stability: > 2 years under recommended storage conditions

Analyte: endo-Cellulase

Assay Format: Spectrophotometer, Auto-analyser

Detection Method: Absorbance

Wavelength (nm): 400

Signal Response: Increase

Limit of Detection: 1.2 x 10-3 U/mL

Reproducibility (%): ~ 3%

Total Assay Time: 10 min

Application examples: Fermentation broths, industrial enzyme preparations and biofuels research.