10 mL; 3,000 U/mL
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Content: | 10 mL; 3,000 U/mL |
Shipping Temperature: | Ambient |
Storage Temperature: | Below -10oC |
Formulation: | In 50% (v/v) glycerol plus 0.02% sodium azide |
Physical Form: | Solution |
Stability: | > 4 years below -10oC |
Enzyme Activity: | α-Amylase |
EC Number: | 3.2.1.1 |
CAZy Family: | GH13 |
CAS Number: |
9000-90-2, 9000-85-5 |
Synonyms: | alpha-amylase; 4-alpha-D-glucan glucanohydrolase |
Source: | Bacillus sp. |
Molecular Weight: | 58,000 |
Concentration: | Supplied at ~ 3,000 U/mL |
Expression: | Purified from Bacillus sp. |
Specificity: | Hydrolysis of α-1,4 glucosidic linkages in linear α-1,4 glucan (e.g. amylose regions in starch). |
Specific Activity: | ~ 170 U/mg (40oC, pH 6.5 on Ceralpha reagent) |
Unit Definition: | One Unit of α-amylase activity is defined as the amount of enzyme required to release one μmole of p-nitrophenol from blocked p-nitrophenyl-maltoheptaoside per minute (in the presence of excess α-glucosidase) at pH 6.5 and 40oC. |
Temperature Optima: | 100oC |
pH Optima: | 7 |
Application examples: | For use in Megazyme Total Starch Assay Kits (K-TSTA and K-TSHK). |
High purity α-Amylase (Thermostable) (Bacillus sp.) for use in research, biochemical enzyme assays and in vitro diagnostic analysis. This enzyme is exceptionally thermostable and is the ideal choice for use in the total starch assay kits (K-TSTA and K-TSHK). It is considerably more thermostable than related enzymes such as the heat stable α-amylase (Bacillus licheniformis, E-BLAAM). Please see the data sheet for experimental stability data.
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Measurement of available carbohydrates, digestible, and resistant starch in food ingredients and products.
McCleary, B. V., McLoughlin, C., Charmier, L. M. J. & McGeough, P. (2019). Cereal Chemistry, 97(1), 114-137.
Background and objectives: The importance of selectively measuring available and unavailable carbohydrates in the human diet has been recognized for over 100 years. The levels of available carbohydrates in diets can be directly linked to major diseases of the Western world, namely Type II diabetes and obesity. Methodology for measurement of total carbohydrates by difference was introduced in the 1880s, and this forms the basis of carbohydrate determination in the United States. In the United Kingdom, a method to directly measure available carbohydrates was introduced in the 1920s to assist diabetic patients with food selection. The aim of the current work was to develop simple, specific, and reliable methods for available carbohydrates and digestible starch (and resistant starch). The major component of available carbohydrates in most foods is digestible starch. Findings: Simple methods for the measurement of rapidly digested starch, slowly digested starch, total digestible starch, resistant starch, and available carbohydrates have been developed, and the digestibility of phosphate cross‐linked starch has been studied in detail. The resistant starch procedure developed is an update of current procedures and incorporates incubation conditions with pancreatic α‐amylase (PAA) and amyloglucosidase (AMG) that parallel those used AOAC Method 2017.16 for total dietary fiber. Available carbohydrates are measured as glucose, fructose, and galactose, following complete and selective hydrolysis of digestible starch, maltodextrins, maltose, sucrose, and lactose to glucose, fructose, and galactose. Sucrose is hydrolyzed with a specific sucrase enzyme that has no action on fructo‐oligosaccharides (FOS). Conclusions: The currently described “available carbohydrates” method together with the total dietary fiber method (AOAC Method 2017.16) allows the measurement of all carbohydrates in food products, including digestible starch. Significance and novelty: This paper describes a simple and specific method for measurement of available carbohydrates in cereal, food, and feed products. This is the first method that provides the correct measurement of digestible starch and sucrose in the presence of FOS. Such methodology is essential for accurate labeling of food products, allowing consumers to make informed decisions in food selection.
Hide AbstractEvaluation of brewer's spent grain hydrolysate as a substrate for production of thermostable α-amylase by Bacillus stearothermophilus.
Ravindran, R., Williams, G. A. & Jaiswal, A. K. (2019). Bioresource Technology Reports, 5, 141-149.
In the present study, BSG was hydrolysed using cellulolytic enzymes and used as a growth medium supplement for cultivation of the thermophilic bacterium, Bacillus stearothermophilus in the production of α-amylase. A central composite design involving five parameters and four levels viz. starch, peptone, KCl, and MgSO4 along with BSG hydrolysate was used to derive the optimal media composition. The fermentation was conducted using shake flasks for 36 h at a temperature of 50°C and pH 7.0 at 220 rpm. Optimization trials revealed that maximal amylase production (198.09 U/ml) occurred with a medium composition of starch (0.2% w/v), peptone (0.2% w/v), KCl·4 H2O (0.02% w/v), MgSO4·7 H2O (0.01% w/v) and hydrolysate (0.22% v/v). A 1.3-fold increase in amylase activity was obtained following novel media composition. All the factors considered in the study were found to be significant. The enzyme was purified by three step purification strategy, characterised and tested for anti-biofilm activity.
Hide Abstract