Improvement of Biocatalytic Properties and Cytotoxic Activity of L-Asparaginase from <i>Rhodospirillum rubrum</i> by Conjugation with Chitosan-Based Cationic Polyelectrolytes
L-asparaginases (L-ASNases, EC 3.5.1.1) are a family of enzymes that are widely used for the treatment of lymphoblastic leukemias. L-ASNase from <i>Rhodospirillum rubrum</i> (RrA) has a low molecular weight, low glutaminase activity, and low immunogenicity, making it a promising enzyme f...
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MDPI AG
2022-03-01
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| author | Natalia V. Dobryakova Dmitry D. Zhdanov Nikolay N. Sokolov Svetlana S. Aleksandrova Marina V. Pokrovskaya Elena V. Kudryashova |
| author_facet | Natalia V. Dobryakova Dmitry D. Zhdanov Nikolay N. Sokolov Svetlana S. Aleksandrova Marina V. Pokrovskaya Elena V. Kudryashova |
| author_sort | Natalia V. Dobryakova |
| collection | DOAJ |
| description | L-asparaginases (L-ASNases, EC 3.5.1.1) are a family of enzymes that are widely used for the treatment of lymphoblastic leukemias. L-ASNase from <i>Rhodospirillum rubrum</i> (RrA) has a low molecular weight, low glutaminase activity, and low immunogenicity, making it a promising enzyme for antitumor drug development. In our work, the complex formation and covalent conjugation of the enzyme with synthetic or natural polycationic polymers was studied. Among non-covalent polyelectrolyte complexes (PEC), polyethyleneimine (PEI) yielded the highest effect on RrA, increasing its activity by 30%. The RrA-PEI complex had increased stability to trypsinolysis, with an inactivation constant decrease up to 10-fold compared to that of the native enzyme. The covalent conjugation of RrA with chitosan-PEI, chitosan-polyethylene glycol (chitosan-PEG), and chitosan-glycol resulted in an increase in the specific activity of L-asparagine (up to 30%). RrA-chitosan-PEG demonstrated dramatically (by 60%) increased cytotoxic activity for human chronic myeloma leukemia K562 cells in comparison to the native enzyme. The antiproliferative activity of RrA and its conjugates was significantly higher (up to 50%) than for that of the commercially available EcA at the same concentration. The results of this study demonstrated that RrA conjugates with polycations can become a promising strategy for antitumor drug development. |
| first_indexed | 2024-03-09T10:30:45Z |
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| language | English |
| last_indexed | 2024-03-09T10:30:45Z |
| publishDate | 2022-03-01 |
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| series | Pharmaceuticals |
| spelling | doaj.art-0c06139e142744e68451b684a8a3e0ff2023-12-01T21:18:01ZengMDPI AGPharmaceuticals1424-82472022-03-0115440610.3390/ph15040406Improvement of Biocatalytic Properties and Cytotoxic Activity of L-Asparaginase from <i>Rhodospirillum rubrum</i> by Conjugation with Chitosan-Based Cationic PolyelectrolytesNatalia V. Dobryakova0Dmitry D. Zhdanov1Nikolay N. Sokolov2Svetlana S. Aleksandrova3Marina V. Pokrovskaya4Elena V. Kudryashova5Chemical Faculty, Lomonosov Moscow State University, Leninskie Gory St. 1, 119991 Moscow, RussiaLaboratory of Medical Biotechnology, Institute of Biomedical Chemistry, Pogodinskaya St. 10/8, 119121 Moscow, RussiaLaboratory of Medical Biotechnology, Institute of Biomedical Chemistry, Pogodinskaya St. 10/8, 119121 Moscow, RussiaLaboratory of Medical Biotechnology, Institute of Biomedical Chemistry, Pogodinskaya St. 10/8, 119121 Moscow, RussiaLaboratory of Medical Biotechnology, Institute of Biomedical Chemistry, Pogodinskaya St. 10/8, 119121 Moscow, RussiaChemical Faculty, Lomonosov Moscow State University, Leninskie Gory St. 1, 119991 Moscow, RussiaL-asparaginases (L-ASNases, EC 3.5.1.1) are a family of enzymes that are widely used for the treatment of lymphoblastic leukemias. L-ASNase from <i>Rhodospirillum rubrum</i> (RrA) has a low molecular weight, low glutaminase activity, and low immunogenicity, making it a promising enzyme for antitumor drug development. In our work, the complex formation and covalent conjugation of the enzyme with synthetic or natural polycationic polymers was studied. Among non-covalent polyelectrolyte complexes (PEC), polyethyleneimine (PEI) yielded the highest effect on RrA, increasing its activity by 30%. The RrA-PEI complex had increased stability to trypsinolysis, with an inactivation constant decrease up to 10-fold compared to that of the native enzyme. The covalent conjugation of RrA with chitosan-PEI, chitosan-polyethylene glycol (chitosan-PEG), and chitosan-glycol resulted in an increase in the specific activity of L-asparagine (up to 30%). RrA-chitosan-PEG demonstrated dramatically (by 60%) increased cytotoxic activity for human chronic myeloma leukemia K562 cells in comparison to the native enzyme. The antiproliferative activity of RrA and its conjugates was significantly higher (up to 50%) than for that of the commercially available EcA at the same concentration. The results of this study demonstrated that RrA conjugates with polycations can become a promising strategy for antitumor drug development.https://www.mdpi.com/1424-8247/15/4/406L-asparaginase<i>Rhodospirillum rubrum</i>catalytic activitycationic polymerspolyelectrolyte complexescovalent conjugates |
| spellingShingle | Natalia V. Dobryakova Dmitry D. Zhdanov Nikolay N. Sokolov Svetlana S. Aleksandrova Marina V. Pokrovskaya Elena V. Kudryashova Improvement of Biocatalytic Properties and Cytotoxic Activity of L-Asparaginase from <i>Rhodospirillum rubrum</i> by Conjugation with Chitosan-Based Cationic Polyelectrolytes Pharmaceuticals L-asparaginase <i>Rhodospirillum rubrum</i> catalytic activity cationic polymers polyelectrolyte complexes covalent conjugates |
| title | Improvement of Biocatalytic Properties and Cytotoxic Activity of L-Asparaginase from <i>Rhodospirillum rubrum</i> by Conjugation with Chitosan-Based Cationic Polyelectrolytes |
| title_full | Improvement of Biocatalytic Properties and Cytotoxic Activity of L-Asparaginase from <i>Rhodospirillum rubrum</i> by Conjugation with Chitosan-Based Cationic Polyelectrolytes |
| title_fullStr | Improvement of Biocatalytic Properties and Cytotoxic Activity of L-Asparaginase from <i>Rhodospirillum rubrum</i> by Conjugation with Chitosan-Based Cationic Polyelectrolytes |
| title_full_unstemmed | Improvement of Biocatalytic Properties and Cytotoxic Activity of L-Asparaginase from <i>Rhodospirillum rubrum</i> by Conjugation with Chitosan-Based Cationic Polyelectrolytes |
| title_short | Improvement of Biocatalytic Properties and Cytotoxic Activity of L-Asparaginase from <i>Rhodospirillum rubrum</i> by Conjugation with Chitosan-Based Cationic Polyelectrolytes |
| title_sort | improvement of biocatalytic properties and cytotoxic activity of l asparaginase from i rhodospirillum rubrum i by conjugation with chitosan based cationic polyelectrolytes |
| topic | L-asparaginase <i>Rhodospirillum rubrum</i> catalytic activity cationic polymers polyelectrolyte complexes covalent conjugates |
| url | https://www.mdpi.com/1424-8247/15/4/406 |
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