Megazyme K-PULLG6 普鲁兰酶/极限醍醐精检测试剂盒

Megazyme K-PULLG6 普鲁兰酶/极限醍醐精检测试剂盒

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Megazyme K-PULLG6 普鲁兰酶/极限醍醐精检测试剂盒

中文名称：普鲁兰酶/极限醍醐精检测试剂盒

英文名：Pullulanase/Limit-Dextrinase Assay Kit (PULLG6 Method)

货号：K-PULLG6

规格：100 assays (manual)

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.

PullG 6法测定普鲁兰酶采用水溶性底物，即4，6-O-苄基-4-硝基苯-63-α-D-麦芽糖-丙三糖(BPNPG3G3)，与α-葡萄糖苷酶和β-葡萄糖苷酶结合.当底物在1，6-α-链上被普鲁兰酶或极限糊精酶水解后，释放出的4-硝基苯基-β-麦芽三糖苷在试剂混合物中α-葡萄糖苷酶和β-葡萄糖苷酶的协同作用下，立即水解成葡萄糖和4-硝基酚。反应结束后，加入稀碱，形成酚酸根离子。在400 nm处读取吸光度，得到的吸光度值与普鲁兰酶活性直接相关。

Colourimetric method for the determination of pullanase
or limit-dextrinase

Principle:
(pullulanase/limit-dextrinase)
(1) Benzylidene-G3-(α-1,6)-G3-β-PNP + H2O → Benzylidene-G3 
+ G3-β-PNP

(thermostable α-glucosidase and β-glucosidase)
(2) G3-β-PNP + H2O → D-glucose + PNP

(alkaline solution)
(3) PNP → phenolate ion (yellow colour)

Note: PNP = 4-nitrophenol

Kit size: 100 assays
Method: Spectrophotometric at 400 nm
Total assay time: 10 min for pullanase preparations
30 min for malt extracts containing
limit-dextrinase
Detection limit: 0.18 U/mL for pullulanase preparations
(50-fold dilution)
0.01 U/g for limit dextrinase in milled malt
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