Method for the measurement of Pullulanase/Limit Dextrinase

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.
he absorbance is read at 400 nm and the value obtained correlates directly with pullulanase activity.

Shipping temperature: ambient

Storage temperature:
- Short term stability: 2-8^oC,
- Long term stability: see individual component labels

Assay format: spectrophotometer

Detection method: absorbance - 400

Signal response: increase 

Total assay time: *10 min for pullanase preparations
                               *30 min for malt extracts containing limit-dextrinase

Detection limit: 18 U/ml for pullulanase preparations (50-fold dilution) and 0,01 U/g for limit dextrinase in milled malt

Reproducibility (%): ~ 3%

Stability: > 2 years under recommended storage condition

• 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

Assay of microbial pullulanase preparations and measurement of limit-dextrinase in malt extracts

Safety data sheet Calculator data Validation report Data Booklet

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