PullG6 assay for the measurement of pullulanase employs a water soluble defined substrate, namely 4,6-O-benzylidene-4-nitrophenyl-63-α-D-maltotriosyl-maltotriose (BPNPG3G3), coupled with the ancillary enzymes α-glucosidase and β-glucosidase. Upon hydrolysis of the substrate at the 1,6-α-linkage by pullulanase or limit-dextrinase, the released 4-nitrophenyl-β-maltotrioside is immediately hydrolysed to glucose and 4-nitrophenol by the concerted action of the α-glucosidase and β-glucosidase enzymes in the reagent mixture. The reaction is terminated and phenolate ions are developed by addition of dilute alkali. The absorbance is read at 400 nm and the value obtained correlates directly with pullulanase activity.

 

Content: 100 / 200 assays per kit

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: Pullulanase/Limit-Dextrinase

Assay Format: Spectrophotometer

Detection Method: Absorbance

Wavelength (nm): 400

Signal Response: Increase

Limit of Detection: 0.18 U/mL for pullulanase preparations (50-fold dilution)

0.01 U/g for limit dextrinase in milled malt

Reproducibility (%): ~ 3%

Total Assay Time: ~ 10 min (Pullanase),

~ 30 min (Limit-Dextrinase)

Application examples: Assay of microbial pullulanase preparations. Measurement of limit-dextrinase in malt extracts.

Method recognition: Novel method

 

Advantages

High sensitivity 

Suitable for manual and auto-analyser formats 

No transglycosylation interference 

Very cost effective 

All reagents stable for > 1 year after preparation 

Very specific

Simple format

Standard included

 

 

用于测定普鲁兰酶的PullG6测定法使用水溶性确定的底物，即4,6-O-亚苄基-4-硝基苯-63-α-D-麦芽三糖基-麦芽三糖酶（BPNPG3G3），与辅助酶α-葡萄糖苷酶和β-葡糖苷酶偶联。在1，6-α-键的底物被普鲁兰酶或极限糊精酶水解后，释放的4-硝基苯基-β-麦芽三糖苷在试剂混合物中的α-葡萄糖苷酶和β-葡萄糖苷酶类的协同作用下立即水解为葡萄糖和4-硝基酚。通过加入稀碱终止反应并产生酚根离子。在400nm处读取吸光度，并且所获得的值与普鲁兰酶活性直接相关。

含量：每个试剂盒100/200次测定

运输温度：环境温度

储存温度：短期稳定性：2-8oC，

长期稳定性：参见单个组件标签

稳定性：在推荐的储存条件下超过2年

分析物：普鲁兰酶/极限糊精

测定形式：分光光度计

检测方法：吸光度

波长（nm）：400

信号响应：增加

检出限：普鲁兰酶制剂为0.18 U/mL（50倍稀释）

研磨麦芽中极限糊精酶为0.01U/g

再现性（%）：~3%

总测定时间：约10分钟（Pullanase），

约30分钟（限制右旋糖苷酶）

应用实例：微生物普鲁兰酶制剂的测定。麦芽提取物中极限糊精酶的测定。

方法识别：一种新颖的方法

优点

高灵敏度

适用于手动和自动分析仪格式

无转糖基化干扰

非常经济高效

所有试剂在制备后稳定超过1年

非常具体

简单格式

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