Amylolytic Enzyme Activity Assessment Service

Efficient Amylolytic Enzymes Activity Assessment Service at Creative Biolabs

Amylolytic enzymes are a class of enzymes that degrade starch and other polysaccharides. For example, α-amylase hydrolyzes the α-1,4-glucosidic bonds within starch molecules, thereby breaking them down into shorter chain-like sugar molecules such as maltose, dextrin, and glucose. β-amylase acts on the non-reducing end of the starch molecule and hydrolyzes it into maltose. The main function of glucoamylase is to hydrolyze the non-reducing terminal α-1,4-glucosidic bond of starch molecules and break them into glucose molecules, and it is an important digestive enzyme and industrial enzyme. These different types of amylolytic enzymes play important roles in food processing, winemaking, biofuels, and other fields, promoting the degradation and utilization of starch. Each type of enzyme works under specific conditions, working together to complete the complete degradation process of starch molecules. Creative Biolabs has rich experience in Enzyme Activity Assessment services. We design appropriate experimental plans and conduct reliable enzyme activity tests according to client needs, such as Cellulases, Cellulolytic Enzymes, and amylolytic enzymes.

We provide high-level analysis for many types of amylolytic enzyme activity through the following steps, including α-amylase, β-amylase, and glucoamylase.

• Sample pretreatment

Before performing amylolytic enzyme activity analysis, sample pretreatment is crucial. We perform appropriate pretreatment of the samples according to the experimental purpose. For example: We centrifuge samples to remove impurities or solid particles. The sample is then diluted to an appropriate concentration range to ensure the accuracy of the experimental results.

• Preparation of enzyme reaction system

We add amylolytic enzyme solution and mix substrate solution (usually containing starch) in appropriate buffers and mix them thoroughly in the same system. We control experimental conditions, such as temperature, pH, and reaction time, to start the catalytic reaction of amylolytic enzymes. The reaction is then terminated by adding an appropriate reaction stop reagent, and the reaction mixture is centrifuged to obtain a clear supernatant or precipitate for subsequent analysis.

• Enzyme activity assay

We evaluate amylolytic enzyme activity by measuring the products generated by the reaction or the substrate consumed. Different methods differ in terms of operational procedures, sensitivity, and quantitative accuracy. Common measurement methods include:

• Photometry: Enzyme activity is assessed by measuring the change in absorbance resulting from substrate conversion.
• Titration: Enzyme activity is measured by measuring the amount of a specific product (such as a sugar) produced in a reaction.
• Chromatography: We use high-performance liquid chromatography, ion chromatography, and other techniques to separate and quantify substrates and products to evaluate enzyme activity.
• Mass spectrometry: Combined with mass spectrometry technology, we accurately identify substrates and products to assess enzyme activity.

Combined with a standard curve or a specific calculation formula, we convert the measured data into amylolytic enzyme activity units (the amount of product released or the amount of substrate consumed per unit time). Then we perform statistical analysis on the experimental results and deliver the analysis results to the client in the form of a report.

Creative Biolabs has experienced and professional teams that accurately perform enzyme activity analysis and interpret the results. We are equipped with advanced laboratory equipment, such as high-performance liquid chromatography, mass spectrometer, etc., to ensure accurate and rapid analysis of amylolytic enzyme activity. Don't hesitate to contact us promptly if you need high-quality amylolytic enzyme activity analysis, we are your best choice.

Published data

α-Amylase is an endoglucanase that works by cleaving α-1,4-glycosidic bonds within starch to produce shorter sugar molecules. Glucoamylase is an exoglucanase that releases the monosaccharide glucose from the non-reducing end of starch. The hydrolytic activity of α- or glucoamylase is generally measured by measuring an increase in the amount of product (reducing sugars) or a decrease in the amount of substrate. The authors customized a protocol for measuring amylolytic enzyme activity. They took advantage of the micro-thermal cycler's ability to accurately and quickly control temperature and conducted micro-measurements on the activities of α-amylase and glucoamylase in the thermal cycler through the Somogyi-Nelson test (SNT) and the starch iodine test (SIT). The results showed that microSIT was more sensitive in detecting endoglucanase activity, while microSNT was more sensitive in detecting exoglucanase activity. This assay scheme reduced reagent and sample volumes, further improved the sensitivity of the assay, and helped develop new biocatalysts and analyzed sugar-containing samples.

Fig.1 Activity evaluation results of glucoamylase (a) and α-amylase (b).1

FAQs

Q1: What is your experience analyzing different types of enzymes? What does your analytics service include?

A1: Our team has extensive experience and is familiar with various types of enzymes and their characteristics, and conducts accurate analysis of different enzymes. For example, we have successfully analyzed various amylolytic enzymes such as α-amylase and glucoamylase. We provide comprehensive amylolytic enzyme activity analysis services, including standard curve preparation, sample processing, absorbance measurement, data analysis, and result interpretation.

Q2: What sample types are your amylolytic enzyme analysis services suitable for?

A2: Our analysis services are suitable for various types of samples, such as purified enzyme samples, enzyme samples extracted from cells or tissues, fermentation broth, cereals, starches, and other sample types.

Q3: How are your experimental conditions controlled?

A3: We strictly control experimental conditions to ensure experimental repeatability and stability. We regularly calibrate instruments, use quality-controlled reagents, and conduct blank control experiments to ensure the accuracy of analysis results.

Customer Review

Reference

1. Borkowska, Monika, et al. "Rapid micro-assays for amylolytic activities determination: customization and validation of the tests." Applied Microbiology and Biotechnology 103 (2019): 2367-2379.

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