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Enzyme Activity Measurement for Lyases

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Lyases are a diverse class of enzymes that catalyze the breaking of chemical bonds by means other than hydrolysis or oxidation, often forming new double bonds or ring structures in the process. Their unique reaction mechanisms make them indispensable tools in metabolic pathways, biotechnology, and industrial biocatalysis. At Creative Enzymes, we provide precise, reliable, and customized enzyme activity measurement services for Lyases, helping clients gain accurate insights into enzyme kinetics, specificity, and functional behavior.

Understanding Lyases Activity Measurement

Lyases play important roles in cellular metabolism by catalyzing the removal of groups from substrates or the addition of groups to double bonds. They are categorized into subclasses such as decarboxylases, dehydratases, synthases, and aldolases, each with unique applications.

Applications of lyase activity measurement include:

  • Metabolic Engineering: Optimizing biosynthetic pathways.
  • Drug Discovery and Pharmacology: Identifying and characterizing enzyme targets.
  • Industrial Biotechnology: Biocatalysis for green chemistry and manufacturing.
  • Environmental Science: Studying enzymes that degrade pollutants or complex biomolecules.

Accurate activity measurement is essential to fully understand their biochemical behavior and harness their potential.

Comprehensive Assay Services

How It Works

Step Procedure Details
1 Enzyme Preparation
  • Handling of natural, recombinant, or engineered lyases.
  • Optional purification to improve measurement consistency.
2 Substrate Selection
  • Careful choice of physiological or analog substrates.
  • Consideration of stability, availability, and relevance to the intended application.
3 Assay Development
  • Customized assay design, depending on the enzyme type and client needs.
  • Available assay formats:
    • Spectrophotometric assays for real-time monitoring.
    • Fluorescent assays for high sensitivity and throughput.
    • Chromatographic methods for precise product identification.
4 Enzyme Activity Determination
  • Quantitative determination of reaction rates under optimized conditions.
  • Real-time or endpoint measurements supported.
5 Data Analysis & Reporting
  • Kinetic analysis, including Km, Vmax, and kcat determination.
  • Expert interpretation of substrate specificity, turnover rates, and inhibition patterns.

Contact Our Team

Why Choose Creative Enzymes

Extensive Expertise

In-depth knowledge of lyase subclasses and reaction mechanisms.

Flexible Assay Formats

Multiple detection platforms for sensitive and accurate measurement.

Customized Solutions

Assays designed to fit specific enzyme properties and project goals.

Reliable Data Quality

Robust protocols and strict quality controls for reproducibility.

Confidential & Secure

Strong protection of client intellectual property and results.

Delivery

Streamlined project workflows ensure rapid turnaround.

Case Studies and Real-World Applications

Case 1: Rational Engineering of Lysine Decarboxylase for Efficient Cadaverine Production

Cadaverine, a key C5 platform compound for polyamides, can be sustainably produced via enzymatic decarboxylation of L-lysine. However, alkaline conditions during production reduce lysine decarboxylase (EcCadA) activity, limiting industrial applications. To address this, rational engineering introduced interfacial disulfide bond mutations (M1, M2, M3) to stabilize the enzyme. Variant M3 (V12C/D41C) showed a sixfold increase in cadaverine production at pH 10.0 by enhancing decamer stability. Fed-batch bioconversion in a 5 L fermenter yielded 418 g/L cadaverine, the highest titer reported. This work highlights an efficient strategy for industrial-scale, eco-friendly cadaverine biosynthesis with reduced acid consumption.

Enhancing pH stability of lysine decarboxylase via rational engineering and its application in cadaverine industrial productionFigure 1. The relative activities of different lysine decarboxylases under different pH conditions. (Gao et al., 2022)

Case 2: Characterization of Fructose-1,6-Bisphosphate Aldolase in Echinococcus multilocularis

Glycolysis is a key energy pathway in Echinococcus multilocularis, with fructose-1,6-bisphosphate aldolase (FBA) playing a central role. Genome-wide analysis identified four FBA genes, each containing the FBA I domain and conserved active sites. EmFBA1 was localized mainly in the germinal layer and posterior protoscolex. Biochemical characterization showed an enzyme activity of 67.42 U/mg, with Km of 1.75 mM and Vmax of 0.5 mmol/min. EmFBA1 activity was strongly inhibited by Fe3+ but unaffected by Na+, Ca2+, K+, or Mg2+. These findings provide new insights into EmFBA1's biochemical properties and its potential role in parasite glycolysis.

Enzyme kinetics of fructose-1,6-bisphosphate aldolase in Echinococcus multilocularisFigure 2. Enzyme kinetics of EmFBA1. (C) The enzyme activity of EmFBA1. The reaction was conducted in 50 mM Tris-HCl (pH 7.5) at 37 °C for 10 min, and absorbance values at 340 nm were recorded at an interval of 1 min. (D) Determination of Km and Vmax of EmFBA1. (He et al., 2021)

FAQs

  • Q: Which subclasses of lyases can you measure?

    A: We measure all types of lyases, including decarboxylases, dehydratases, aldolases, synthases, and more.

  • Q: What types of assays do you provide?

    A: Depending on your enzyme and research goals, we offer spectrophotometric, fluorescent, and chromatographic assays.

  • Q: Can you provide kinetic data in addition to activity measurements?

    A: Yes, we provide full kinetic profiling, including Km, Vmax, kcat, and inhibitor studies if required.

  • Q: Do you work with crude extracts or only purified enzymes?

    A: Both. We can measure enzyme activity in crude extracts, partially purified samples, or fully purified enzymes.

  • Q: How long does a typical lyase activity measurement project take?

    A: Most projects are completed within 2–4 weeks, depending on enzyme complexity and assay requirements.

  • Q: Do you customize assays for unique hydrolase applications?

    A: Absolutely. Each hydrolase behaves differently, so we design customized assays to match the enzyme's natural substrate and your research objectives, ensuring results are both relevant and accurate.

References:

  1. Gao S, Zhang A, Ma D, et al. Enhancing pH stability of lysine decarboxylase via rational engineering and its application in cadaverine industrial production. Biochemical Engineering Journal. 2022;186:108548. doi:10.1016/j.bej.2022.108548
  2. He X, Zhang J, Sun Y, et al. Characterization of Fructose-1,6-Bisphosphate Aldolase 1 of Echinococcus multilocularis. Veterinary Sciences. 2021;9(1):4. doi:10.3390/vetsci9010004
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For research and industrial use only, not for personal medicinal use.

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For research and industrial use only, not for personal medicinal use.