Megazyme K-PULLG6 普魯蘭酶/極限醍醐精檢測試劑盒

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Megazyme K-PULLG6 普魯蘭酶/極限醍醐精檢測試劑盒

中文名稱：普魯蘭酶/極限醍醐精檢測試劑盒

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

貨號：K-PULLG6

規(guī)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法測定普魯蘭酶采用水溶性底物，即4，6-O-芐基-4-硝基苯-6³-α-D-麥芽糖-丙三糖(BPNPG3G3)，與α-葡萄糖苷酶和β-葡萄糖苷酶結(jié)合.當?shù)孜镌?，6-α-鏈上被普魯蘭酶或極限糊精酶水解后，釋放出的4-硝基苯基-β-麥芽三糖苷在試劑混合物中α-葡萄糖苷酶和β-葡萄糖苷酶的協(xié)同作用下，立即水解成葡萄糖和4-硝基酚。反應結(jié)束后，加入稀堿，形成酚酸根離子。在400 nm處讀取吸光度，得到的吸光度值與普魯蘭酶活性直接相關(guān)。

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

Megazyme是分析試劑，酶和檢測試劑盒的全球供應商，Megazyme能夠為世界各地的實驗室開發(fā)，制造和供應產(chǎn)品。 我們的產(chǎn)品已應用于各種行業(yè)，包括食品和飲料，生物燃料，動物飼料，葡萄酒和釀造，乳制品和谷物。
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