Effect of Pollyallylamine on Alcoholdehydrogenase Structure and Activity
In this article, the effect of polyallylamine (PAA) on the structure and catalytic characteristics of alcohol dehydrogenase (ADH) was studied. For this research, we used methods of stationary kinetics and fluorescence spectroscopy. It has been shown that PAA non-competitively inhibits ADH activity w...
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MDPI AG
2020-04-01
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| Series: | Polymers |
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| Online Access: | https://www.mdpi.com/2073-4360/12/4/832 |
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| author | Aleksandr L. Kim Egor V. Musin Alexey V. Dubrovskii Sergey A. Tikhonenko |
| author_facet | Aleksandr L. Kim Egor V. Musin Alexey V. Dubrovskii Sergey A. Tikhonenko |
| author_sort | Aleksandr L. Kim |
| collection | DOAJ |
| description | In this article, the effect of polyallylamine (PAA) on the structure and catalytic characteristics of alcohol dehydrogenase (ADH) was studied. For this research, we used methods of stationary kinetics and fluorescence spectroscopy. It has been shown that PAA non-competitively inhibits ADH activity while preserving its quaternary structure. It was established that 0.1 M ammonium sulfate removes the inhibitory effect of PAA on ADH, which is explained by the binding of sulfate anion (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub> with polyallylamine amino groups. As a result, the rigidity of the polymer chain increases and the ability to bind to the active loop of the enzyme increases. It is also shown that sodium chloride removes the inhibitory effect of PAA on ADH due to an electrostatic screening of the enzyme from polyelectrolyte. The method of encapsulating ADH in polyelectrolyte microcapsules was adapted to the structure and properties of the enzyme molecule. It was found that the best for ADH is its encapsulation by adsorption into microcapsules already formed on CaCO<sub>3</sub> particles. It was shown that the affinity constant of encapsulated alcohol dehydrogenase to the substrate is 1.7 times lower than that of the native enzyme. When studying the affinity constant of ADH in a complex with PAA to ethanol, the effect of noncompetitive inhibition of the enzyme by polyelectrolyte was observed. |
| first_indexed | 2024-03-10T20:39:02Z |
| format | Article |
| id | doaj.art-997950fcbbb3493b83e4a2eb40008098 |
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| issn | 2073-4360 |
| language | English |
| last_indexed | 2024-03-10T20:39:02Z |
| publishDate | 2020-04-01 |
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| series | Polymers |
| spelling | doaj.art-997950fcbbb3493b83e4a2eb400080982023-11-19T20:48:38ZengMDPI AGPolymers2073-43602020-04-0112483210.3390/polym12040832Effect of Pollyallylamine on Alcoholdehydrogenase Structure and ActivityAleksandr L. Kim0Egor V. Musin1Alexey V. Dubrovskii2Sergey A. Tikhonenko3Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya st., 3, Puschino 142290, Moscow Reg., RussiaInstitute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya st., 3, Puschino 142290, Moscow Reg., RussiaInstitute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya st., 3, Puschino 142290, Moscow Reg., RussiaInstitute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya st., 3, Puschino 142290, Moscow Reg., RussiaIn this article, the effect of polyallylamine (PAA) on the structure and catalytic characteristics of alcohol dehydrogenase (ADH) was studied. For this research, we used methods of stationary kinetics and fluorescence spectroscopy. It has been shown that PAA non-competitively inhibits ADH activity while preserving its quaternary structure. It was established that 0.1 M ammonium sulfate removes the inhibitory effect of PAA on ADH, which is explained by the binding of sulfate anion (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub> with polyallylamine amino groups. As a result, the rigidity of the polymer chain increases and the ability to bind to the active loop of the enzyme increases. It is also shown that sodium chloride removes the inhibitory effect of PAA on ADH due to an electrostatic screening of the enzyme from polyelectrolyte. The method of encapsulating ADH in polyelectrolyte microcapsules was adapted to the structure and properties of the enzyme molecule. It was found that the best for ADH is its encapsulation by adsorption into microcapsules already formed on CaCO<sub>3</sub> particles. It was shown that the affinity constant of encapsulated alcohol dehydrogenase to the substrate is 1.7 times lower than that of the native enzyme. When studying the affinity constant of ADH in a complex with PAA to ethanol, the effect of noncompetitive inhibition of the enzyme by polyelectrolyte was observed.https://www.mdpi.com/2073-4360/12/4/832polyallylaminealcohol dehydrogenaseNaCl(NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>alcohol dehydrogenase structurecatalytic characteristics |
| spellingShingle | Aleksandr L. Kim Egor V. Musin Alexey V. Dubrovskii Sergey A. Tikhonenko Effect of Pollyallylamine on Alcoholdehydrogenase Structure and Activity Polymers polyallylamine alcohol dehydrogenase NaCl (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub> alcohol dehydrogenase structure catalytic characteristics |
| title | Effect of Pollyallylamine on Alcoholdehydrogenase Structure and Activity |
| title_full | Effect of Pollyallylamine on Alcoholdehydrogenase Structure and Activity |
| title_fullStr | Effect of Pollyallylamine on Alcoholdehydrogenase Structure and Activity |
| title_full_unstemmed | Effect of Pollyallylamine on Alcoholdehydrogenase Structure and Activity |
| title_short | Effect of Pollyallylamine on Alcoholdehydrogenase Structure and Activity |
| title_sort | effect of pollyallylamine on alcoholdehydrogenase structure and activity |
| topic | polyallylamine alcohol dehydrogenase NaCl (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub> alcohol dehydrogenase structure catalytic characteristics |
| url | https://www.mdpi.com/2073-4360/12/4/832 |
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