100 assays (manual) / 400 assays (auto-analyser)
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(Malt β-glucanase) 100 assays (manual) / 400 (auto-analyser) Or
(Lichenase) 100 / 200 assays (manual) / 330 (auto-analyser)
Short term stability: 2-8oC,
Long term stability: See individual component labels
|Stability:||> 2 years under recommended storage conditions|
|Assay Format:||Spectrophotometer, Auto-analyser|
|Limit of Detection:||
(Malt β-glucanase) 4.3 x 10-4 U/mL Or
(Lichenase) 9.1 x 10-5 U/mL
|Reproducibility (%):||~ 3%|
|Total Assay Time:||
(Malt β-glucanase) ~ 20 min Or
(Lichenase) ~ 10 min
|Application examples:||Crude malt extracts, industrial enzyme preparations.|
|Method recognition:||Novel method|
The MBG4 reagent contains a single substrate, namely 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-(31-β-D-cellotriosyl-glucoside) (BCNPBG4). The benzylidene acetal group prevents any hydrolytic action by exo-acting hydrolytic enzymes such as β-glucosidase or cellobiohydrolase.
Mixed linkage β-glucanase (endo-1,3:1,4-β-glucanase) / lichenase (EC 184.108.40.206) acts specifically to release 2-chloro-4-nitrophenol (CNP) from this substrate. The rate of release of CNP is directly related to the β-glucanase/lichenase activity in a sample. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH = 10.0).
Note that the substrate is not hydrolysed by β-glucosidase or cellobiohydrolase. The substrate can be hydrolysed by certain endo-cellulases (e.g. Trichoderma sp.) but this does not result in an increase in absorbance.
Data calculators are located in the Documents tab.
- Very cost effective
- All reagents stable for > 2 years
- Specific for endo-1,3:1,4-β-glucanase/lichenase
- Simple, convenient, rapid assay
- Well suited to automation
- Malt flour standard and lichenase standard included
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, In Press.
Colourimetric assays were used to measure the activities of six key hydrolases endogenous to barley: β‐glucanase, xylanase, cellulase, α-amylase, beta‐amylase and limit dextrinase. The analysed barley malt samples were previously characterised by 27 conventional malt quality descriptors. Correlations between enzymatic activities and brewing parameters such as extract yield, fermentability, viscosity and filterability were investigated. A single extraction protocol for all six hydrolases was optimised and used for multi‐enzyme analysis using fully automatable assay formats. A regression analysis between malt parameters was undertaken to produce a relationship matrix linking enzyme activities and conventional malt quality descriptors. This regression analysis was used to inform a multi‐linear regression approach to create predictive models for extract yield, apparent attenuation limit, viscosity and filterability using the Small‐scale Wort rapId Filtration Test (SWIFT) and two different mashing protocols – Congress and a modified infusion mash at 65oC (MIM 65oC). It was observed that malt enzyme activities displayed significant correlations with the analysed brewing parameters. Both starch hydrolases and cell wall hydrolase activities together with modification parameters (i.e. Kolbach index) were found to be highly correlated with extract yield and apparent attenuation limit. Interestingly, it was observed that xylanase activity in malts was an important predictor for wort viscosity and filterability. It is envisaged that the automatable measurement of enzyme activity could find use in plant breeding progeny selection and for routine assessment of the functional brewing performance of malt batches. This analytical approach would also contribute to brewing process consistency, product quality and reduced processing times.Hide Abstract
Mangan, D., Liadova, A., Ivory, R. & McCleary, B. V. (2016). Carbohydrate Research, 435, 162-172.
We report herein the development of a novel assay procedure for the measurement of β-glucanase and lichenase (EC 220.127.116.11) in crude enzyme extracts. Two assay formats based on a) a direct cleavage or b) an enzyme coupled substrate were initially investigated. The ‘direct cleavage’ substrate, namely 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-31-cellotriosyl-β-glucopyranoside (MBG4), was found to be the more generally applicable reagent. This substrate was fully characterised using a crude malt β-glucanase extract, a bacterial lichenase (Bacillus sp.) and a non-specific endo-1,3(4)-β-glucanase from Clostridium thermocellum (EC 18.104.22.168). Standard curves were derived that allow the assay absorbance response to be directly converted to β-glucanase/lichenase activity on barley β-glucan. The specificity of MBG4 was confirmed by analysing the action of competing glycosyl hydrolases that are typically found in malt on the substrate. Manual and automated assay formats were developed for the analysis of a) β-glucanase in malt flour and b) lichenase enzyme extracts and the repeatability of these assays was fully investigated.Hide Abstract