Email this article DETERMINATION OF BIOCHEMICAL ACTIVITY OF ARGININE DEIMINASE
- Authors: Egidarova E.1, Grudinina N.A.2, Zabrodskaya Y.A.3,4,5, Suvorov A.N.6, Sokolov A.V.7
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Affiliations:
- Federal State Budgetary Scientific Institution «Institute of Experimental Medicine»
- Institute of Experimental Medicine RAS
- Smorodintsev Research Institute of Influenza
- Petersburg Nuclear Physics Institute named by B.P. Konstantinov of NRC “Kurchatov Institute”
- Peter the Great Saint Petersburg Polytechnic University
- Institute of Experimental Medicine
- Smorodintsev Research Institute of Influenza Peter the Great St Petersburg University Federal State Budgetary Scientific Institution "Institut of Experimental Medicine"
- Section: Original research
- Published: 17.10.2025
- URL: https://journals.eco-vector.com/MAJ/article/view/677017
- DOI: https://doi.org/10.17816/MAJ677017
- ID: 677017
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Abstract
The creation of new antitumor drugs remains relevant. Targeted drugs are especially in demand at present. It is advisable to obtain this enzyme in a recombinant form, active and soluble, taking into account the cytotoxic properties of arginine deiminase.
Objective: To obtain purified, active recombinant arginine deiminase and biochemically evaluate its properties.
Materials and methods: A method for determining the activity of arginine deiminase based on a modified Sakaguchi reaction was developed. Recombinant arginine deiminase was obtained by cloning of arcA gene in E. coli cells. Also a new sorbent for metal-chelate chromatography was developed for purifying the recombinant enzyme.
Results: Cloning of the arcA gene resulted in purified, soluble, and enzymatically active rADI. It is necessary to maintain a pH of about 6,6 to maintain enzyme activity during purification. The active enzyme (specific activity – 25 micromole of Arg per minute per mg of protein) was obtained through the use of a new variant of metal chelate chromatography. It was determined that to preserve the frozen enzyme, it is necessary to add NaCl to a final concentration of 1 M and 2-mercaptoethanol to a final concentration of 0,5 mM.
Conclusion: The recombinant enzyme, freed from LPS impurities, can be used to determine its cytotoxicity on various cancer cell lines, as well as in in vivo experiments on laboratory animal models. The revealed high enzymatic activity of arginine deiminase in vitro makes further research promising for the creation of a new class of antitumor drugs.
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About the authors
Elena Egidarova
Federal State Budgetary Scientific Institution «Institute of Experimental Medicine»
Email: lena.egidarova.97@mail.ru
ORCID iD: 0000-0002-5716-0836
SPIN-code: 3449-0970
Researcher, Department of Molecular Biology, Genetics and Fundamental Medicine Russian Federation
Natalia A. Grudinina
Institute of Experimental Medicine RAS
Email: strangecatnap@gmail.com
ORCID iD: 0000-0001-8483-8967
SPIN-code: 4304-3410
Cand. of Biol. Sci., senior researcher, Department of Molecular Genetics
Russian Federation, 12, Academic Pavlov street, Saint-Petersburg, 197376Yana A. Zabrodskaya
Smorodintsev Research Institute of Influenza; Petersburg Nuclear Physics Institute named by B.P. Konstantinov of NRC “Kurchatov Institute”; Peter the Great Saint Petersburg Polytechnic University
Email: zabryaka@yandex.ru
ORCID iD: 0000-0003-2012-9461
SPIN-code: 3907-8702
PhD in Physic, Researcher of the Department of Molecular Biology of Viruses
Russian Federation, St. PetersburgAlexander N. Suvorov
Institute of Experimental Medicine
Email: alexander_suvorov1@hotmail.com
ORCID iD: 0000-0003-2312-5589
SPIN-code: 8062-5281
Scopus Author ID: 7101829979
ResearcherId: J-6921-2013
MD, Dr. Sci. (Med.), Professor, Corresponding Member of the Russian Academy of Sciences, Head of the Department of Molecular Microbiology
Russian Federation, Saint PetersburgAlexey V. Sokolov
Smorodintsev Research Institute of InfluenzaPeter the Great St Petersburg University
Federal State Budgetary Scientific Institution "Institut of Experimental Medicine"
Author for correspondence.
Email: biochemsokolov@gmail.com
ORCID iD: 0000-0001-9033-0537
SPIN-code: 7427-7395
Doctor of Biological Sciences, Researcher, Laboratory of Systems Virology Department of Molecular Biology of Viruses
Federal State Budgetary Institution "A.A. Smorodintsev Research Institute of Influenza" of the Ministry of Health of the Russian Federation
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