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Enzyme Discovery by Direct Activity Measurement

About Enzyme Discovery by Direct Activity Measurement

Discovery of new enzymes by direct measurement of enzyme activity is a method for screening and discovering new enzymes based on their catalytic activity. Unlike traditional enzyme discovery methods based on gene or protein sequences, it enables rapid discovery of potential enzyme activities, including unknown enzymes or enzymes that have not yet been characterized. However, it requires ultra-high-throughput activity measurement capabilities and strong method development skills. This strategy is useful when specific enzymatic activity is discovered in a certain type of natural sample but without clear parameters such as substrate specificity or enantioselectivity. For example, novel amino acid oxidases can be screened for substrate specificity with enriched culture of microorganism samples. Microorganisms such as bacteria and yeasts are easier to handle than other higher organisms. For cultivable microorganisms, high-throughput microbial screening for enzymes can be carried out with colony pickers and robots, followed by biochemical detection of the enzyme activities. Also, various selective isolation technologies are applied, such as the "enrichment culture technique" and "acclimation technique". In one study, FAD-dependent L-arginine oxidase specific to L-arginine was discovered in newly isolated Pseudomonas sp. TPU 7192 with a simple enzymatic method. After triple-enrichment cultures, the selected strain was identified with genomic DNA sequencing, and the activity was evaluated spectrophotometrically by measuring the rate of hydrogen peroxide formation. In another example, enzymes were extracted from plant seeds. HPLC determined the activity and the stereochemistry.

Service Content for Enzyme Discovery by Direct Activity Measurement

We provide professional services for enzyme discovery by direct activity measurement and are committed to providing efficient, accurate, and reliable enzyme discovery solutions for our customers. With state-of-the-art high-throughput enzyme screening platforms and equipment, as well as a team of experienced scientists, we can utilize various enzyme activity measurement technologies for the rapid discovery of new enzymes. Our specialized one-stop services include but are not limited to:

  • Natural sample enrichment and identification.
  • Enzyme extraction in the native and active state.
  • High-throughput activity screening including method development.
  • Enzyme isolation and purification strategy development.
  • Activity, specificity, and structure characterization of new enzymes.
  • Amino acid analysis and secondary structure prediction.

Sample range: It includes but is not limited to samples from different natural sources such as soil microorganisms, plants, and animals.

Range of target enzymes: It includes but is not limited to hydrolases, oxidoreductases, and lytic enzymes.

Activity assay: High-throughput activity assay.

Service Flow for Enzyme Discovery by Direct Activity Measurement

  1. Project consultation and planning: Understanding your enzyme discovery needs and goals, we develop a direct activity measurement protocol that fits your project based on key factors such as sample type, reaction conditions, substrate selection, and assay method.
  2. Sample preparation: You can provide the samples or we can handle and prepare the samples according to your requirements.
  3. Direct activity measurement experiments: We will use advanced enzyme activity measurement techniques to measure the catalytic activity of the sample under appropriate reaction conditions. These techniques include absorbance measurements, fluorescence measurements, mass spectrometry, etc., and are selected according to different enzyme types and substrate properties.
  4. Data analysis and interpretation: We perform a comprehensive data analysis of the direct activity measurements, accompanied by detailed reports and graphs, and provide explanations and recommendations on the activity of the discovered enzymes, enabling you to gain insight into the properties and potential of the discovered enzymes.

With our direct activity measurement for enzyme discovery service, you can quickly obtain accurate and reliable enzyme activity data to analyze enzyme function and catalytic mechanism, providing strong support for your research and applications. Whether you are engaged in academic research or biopharmaceutical development, we can provide you with the best quality experimental services. Please contact our team to learn more about our services and solutions.

References:

  1. [1] Matsui, D., Terai, A., Asano, Y. (2016) L-Arginine oxidase from Pseudomonas sp. TPU 7192: Characterization, gene cloning, heterologous expression, and application to L-arginine determination. Enzyme and Microbial Technology. 82: 151-157.
  2. [2] Asano, Y., Tamura, K., Doi, N., Ueatrongchit, T., H-Kittikun, A., Ohmiya, T. (2005) Screening for new hydroxynitrilases from plants. Bioscience, Biotechnology and Biochemistry. 69: 2349-2357.

Novel enzymes identification from environmental sample Figure 1. Representation of steps leading to the identification of novel enzymes.
(Biotechnology, Agronomy, Society and Environment 2016)