Pullulanase/Limit-Dextrinase Assay Kit (PullG6 Method)

- Overview
  - 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 400nm and the value obtained correlates directly with pullulanase activity.
    
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- Properties
  - Details
    
    Limit of Detection: 0.18U/mL for pullulanase preparations (50-fold dilution) 0.01U/g for limit dextrinase in milled malt
    
    Storage
    
    Short term stability: 2-8°C
    Long term stability: See individual component labels
    
    Stability
    
    > 2 years under recommended storage conditions
    
    * For research use only.
- Applications
  - Application
    
    Activity Assay
    
    Application Description
    
    Assay of microbial pullulanase preparations. Measurement of limit-dextrinase in malt extracts.
- Reference
  - 1.Diastatic power and maltose value: a method for the measurement of amylolytic enzymes in malt. Charmier, L. M., McLoughlin, C. & McCleary, B. V. (2021). Journal of the Institute of Brewing, In Press.
    2.Prediction of potential malt extract and beer filterability using conventional and novel malt assays. Cornaggia, C., Evans, D. E., Draga, A., Mangan, D. & McCleary, B. V. (2019). Journal of Institute of Brewing, 125(3), 294-309.
    3.Colourimetric and fluorimetric substrates for the assay of limit dextrinase. Mangan, D., McCleary, B. V., Cornaggia, C., Ivory, R., Rooney, E. & McKie, V. (2015). Journal of Cereal Science, 62, 50-57.
    4.Colourimetric and fluorometric substrates for measurement of pullulanase activity. McCleary, B. V., Mangan, D., McKie, V., Cornaggia, C., Ivory, R. & Rooney, E. (2014). Carbohydrate Research, 393, 60-69.
    5.A quick and simple gel diffusion assay to visualize and quantify pullulanase activity for resistant starch content in food crops. Krishnan, V., Awana, M., Kulshreshta, A., Praveen, S. & Singh, A. (2022). Journal of Plant Biochemistry and Biotechnology, 1-7.
    6.Effects of post-heading high temperature on some quality traits of malt barley. Ni, S. J., Zhao, H. F. & Zhang, G. P. (2020). Journal of Integrative Agriculture, 19(11), 2674-2679.
    7.High amylose wheat starch structures display unique fermentability characteristics, microbial community shifts and enzyme degradation profiles. Bui, A., Williams, B., Hoedt, E., Morrison, M., Mikkelsen, D. & Gidley, M. (2020). Food & Function, 6.
    8.Changes in malt quality during production in two commercial malt houses. Yousif, A. M. & Evans, D. E. (2020). Journal of the Institute of Brewing, 126 (3), 233-252.
    9.The influence of drought stress on malt quality traits of the wild and cultivated barleys. Hong, Y. & Zhang, G. P. (2020). Journal of Integrative Agriculture, 19(8), 2009-2015.
    10.NTRC and Thioredoxin f Overexpression Differentially Induces Starch Accumulation in Tobacco Leaves. Ancín, M., Larraya, L., Millán, F. S., Veramendi, J., Burch-Smith, T. & Farran, I. (2019). Plants, 8(12), 543.
    11.Gene Expression Profiling in Short‐Term Imbibition of Wheat: Tools for Dissecting of Pasting Properties of Imbibed Wheat Seeds. Tamura, T., Akuzawa, S. & Mura, K. (2019). Journal of Food Science, 84(5), 946-953.
    12.Rice malting optimization for the production of top‐fermented gluten‐free beer. Ceccaroni, D., Marconi, O., Sileoni, V., Wray, E. & Perretti, G. (2019). Journal of the Science of Food and Agriculture, 99(6), 2726-2734.
    13.Gluten-free sources of fermentable extract: effect of temperature and germination time on quality attributes of teff [Eragrostis tef (zucc.) trotter] malt and wort. Di Ghionno, L., Marconi, O., Lee, E. G., Marconi, O., Rice, C. J., Sileoni, V. & Perretti, G. (2017). Journal of Agricultural and Food Chemistry, 65(23), 4777-4785.
    14.Optimization of the production of an extracellular and thermostable amylolytic enzyme by Thermus thermophilus HB8 and basic characterization. Akassou, M. & Groleau, D. (2017). Extremophiles, 1-14.

MSDS

Pullulanase/Limit-Dextrinase Assay Kit (PullG6 Method)

Pullulanase/Limit-Dextrinase Assay Kit (PullG6 Method)

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