Improving the Efficiency of Electrocatalysis of Cytochrome P450 3A4 by Modifying the Electrode with Membrane Protein Streptolysin O for Studying the Metabolic Transformations of Drugs
In the present work, screen-printed electrodes (SPE) modified with a synthetic surfactant, didodecyldimethylammonium bromide (DDAB) and streptolysin O (SLO) were prepared for cytochrome P450 3A4 (CYP3A4) immobilization, direct non-catalytic and catalytic electrochemistry. The immobilized CYP3A4 demo...
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MDPI AG
2023-04-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/13/4/457 |
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| author | Polina I. Koroleva Andrei A. Gilep Sergey V. Kraevsky Tatiana V. Tsybruk Victoria V. Shumyantseva |
| author_facet | Polina I. Koroleva Andrei A. Gilep Sergey V. Kraevsky Tatiana V. Tsybruk Victoria V. Shumyantseva |
| author_sort | Polina I. Koroleva |
| collection | DOAJ |
| description | In the present work, screen-printed electrodes (SPE) modified with a synthetic surfactant, didodecyldimethylammonium bromide (DDAB) and streptolysin O (SLO) were prepared for cytochrome P450 3A4 (CYP3A4) immobilization, direct non-catalytic and catalytic electrochemistry. The immobilized CYP3A4 demonstrated a pair of redox peaks with a formal potential of −0.325 ± 0.024 V (vs. the Ag/AgCl reference electrode). The electron transfer process showed a surface-controlled mechanism (“protein film voltammetry”) with an electron transfer rate constant (k<sub>s</sub>) of 0.203 ± 0.038 s<sup>−1</sup>. Electrochemical CYP3A4-mediated reaction of N-demethylation of erythromycin was explored with the following parameters: an applied potential of −0.5 V and a duration time of 20 min. The system with DDAB/SLO as the electrode modifier showed conversion of erythromycin with an efficiency higher than the electrode modified with DDAB only. Confining CYP3A4 inside the protein frame of SLO accelerated the enzymatic reaction. The increases in product formation in the reaction of the electrochemical N-demethylation of erythromycin for SPE/DDAB/CYP3A4 and SPE/DDAB/SLO/CYP3A4 were equal to 100 ± 22% and 297 ± 7%, respectively. As revealed by AFM images, the SPE/DDAB/SLO possessed a more developed surface with protein cavities in comparison with SPE/DDAB for the effective immobilization of the CYP3A4 enzyme. |
| first_indexed | 2024-03-11T05:12:24Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-11T05:12:24Z |
| publishDate | 2023-04-01 |
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| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-c8364465c1f2433e91bdc428db511bfb2023-11-17T18:30:57ZengMDPI AGBiosensors2079-63742023-04-0113445710.3390/bios13040457Improving the Efficiency of Electrocatalysis of Cytochrome P450 3A4 by Modifying the Electrode with Membrane Protein Streptolysin O for Studying the Metabolic Transformations of DrugsPolina I. Koroleva0Andrei A. Gilep1Sergey V. Kraevsky2Tatiana V. Tsybruk3Victoria V. Shumyantseva4Institute of Biomedical Chemistry, Pogodinskaya Street, 10, Build 8, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Street, 10, Build 8, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Street, 10, Build 8, 119121 Moscow, RussiaInstitute of Bioorganic Chemistry of the National Academy of Sciences of Belarus, 220141 Minsk, BelarusInstitute of Biomedical Chemistry, Pogodinskaya Street, 10, Build 8, 119121 Moscow, RussiaIn the present work, screen-printed electrodes (SPE) modified with a synthetic surfactant, didodecyldimethylammonium bromide (DDAB) and streptolysin O (SLO) were prepared for cytochrome P450 3A4 (CYP3A4) immobilization, direct non-catalytic and catalytic electrochemistry. The immobilized CYP3A4 demonstrated a pair of redox peaks with a formal potential of −0.325 ± 0.024 V (vs. the Ag/AgCl reference electrode). The electron transfer process showed a surface-controlled mechanism (“protein film voltammetry”) with an electron transfer rate constant (k<sub>s</sub>) of 0.203 ± 0.038 s<sup>−1</sup>. Electrochemical CYP3A4-mediated reaction of N-demethylation of erythromycin was explored with the following parameters: an applied potential of −0.5 V and a duration time of 20 min. The system with DDAB/SLO as the electrode modifier showed conversion of erythromycin with an efficiency higher than the electrode modified with DDAB only. Confining CYP3A4 inside the protein frame of SLO accelerated the enzymatic reaction. The increases in product formation in the reaction of the electrochemical N-demethylation of erythromycin for SPE/DDAB/CYP3A4 and SPE/DDAB/SLO/CYP3A4 were equal to 100 ± 22% and 297 ± 7%, respectively. As revealed by AFM images, the SPE/DDAB/SLO possessed a more developed surface with protein cavities in comparison with SPE/DDAB for the effective immobilization of the CYP3A4 enzyme.https://www.mdpi.com/2079-6374/13/4/457cytochrome P450 3A4electrochemical analysisstreptolysin Oerythromycinbioreactorelectron transfer |
| spellingShingle | Polina I. Koroleva Andrei A. Gilep Sergey V. Kraevsky Tatiana V. Tsybruk Victoria V. Shumyantseva Improving the Efficiency of Electrocatalysis of Cytochrome P450 3A4 by Modifying the Electrode with Membrane Protein Streptolysin O for Studying the Metabolic Transformations of Drugs Biosensors cytochrome P450 3A4 electrochemical analysis streptolysin O erythromycin bioreactor electron transfer |
| title | Improving the Efficiency of Electrocatalysis of Cytochrome P450 3A4 by Modifying the Electrode with Membrane Protein Streptolysin O for Studying the Metabolic Transformations of Drugs |
| title_full | Improving the Efficiency of Electrocatalysis of Cytochrome P450 3A4 by Modifying the Electrode with Membrane Protein Streptolysin O for Studying the Metabolic Transformations of Drugs |
| title_fullStr | Improving the Efficiency of Electrocatalysis of Cytochrome P450 3A4 by Modifying the Electrode with Membrane Protein Streptolysin O for Studying the Metabolic Transformations of Drugs |
| title_full_unstemmed | Improving the Efficiency of Electrocatalysis of Cytochrome P450 3A4 by Modifying the Electrode with Membrane Protein Streptolysin O for Studying the Metabolic Transformations of Drugs |
| title_short | Improving the Efficiency of Electrocatalysis of Cytochrome P450 3A4 by Modifying the Electrode with Membrane Protein Streptolysin O for Studying the Metabolic Transformations of Drugs |
| title_sort | improving the efficiency of electrocatalysis of cytochrome p450 3a4 by modifying the electrode with membrane protein streptolysin o for studying the metabolic transformations of drugs |
| topic | cytochrome P450 3A4 electrochemical analysis streptolysin O erythromycin bioreactor electron transfer |
| url | https://www.mdpi.com/2079-6374/13/4/457 |
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