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Enzyme Discovery by Direct Activity Measurement

Detection of desired enzymes by directly measuring activities in natural sources is a promising method of novel enzyme discovery. The principle is straightforward as suggested by the name, but it quires a large capacity of activity measurement and strong skills in method development, which is the reason that Creative Enzymes is one of the few companies providing such service.

This strategy is useful when specific enzymatic activity was discovered in a certain type of natural samples, 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. Compared with other higher organisms, microorganisms such as bacteria and yeasts are easier to handle. For cultivable microorganisms, high-throughput microbial screening for enzymes can be carried out with colony picker and robots, followed by biochemical detection of the enzyme activities. Also, various selective isolation technologies are applied, such as “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. The activity and the stereochemistry were determined by HPLC.

Creative Enzymes has extensive experience on discovering novel enzymes from different natural sources, such as soil microbes, plants and animals, based on their activities. We have the knowledge to establish high-throughput activity screening for various target enzymes, such as hydrolases, oxidoreductases, and lyases. With our top-tier capacity and years of professional experience in enzyme discovery, we offer professional one-stop services:

References

  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. 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.

Related sections

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



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