The Protein Engineering of Zearalenone Hydrolase Results in a Shift in the pH Optimum of the Relative Activity of the Enzyme
An acidic shift in the pH profile of <i>Clonostachys rosea</i> zearalenone hydrolase (ZHD), the most effective and well-studied zearalenone-specific lactone hydrolase, is required to extend the range of applications for the enzyme as a decontamination agent in the feed and food productio...
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MDPI AG
2024-12-01
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| author | Anna Dotsenko Igor Sinelnikov Ivan Zorov Yury Denisenko Aleksandra Rozhkova Larisa Shcherbakova |
| author_facet | Anna Dotsenko Igor Sinelnikov Ivan Zorov Yury Denisenko Aleksandra Rozhkova Larisa Shcherbakova |
| author_sort | Anna Dotsenko |
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| description | An acidic shift in the pH profile of <i>Clonostachys rosea</i> zearalenone hydrolase (ZHD), the most effective and well-studied zearalenone-specific lactone hydrolase, is required to extend the range of applications for the enzyme as a decontamination agent in the feed and food production industries. Amino acid substitutions were engineered in the active center of the enzyme to decrease the pKa values of the catalytic residues E126 and H242. The T216K substitution provided a shift in the pH optimum by one unit to the acidic region, accompanied by a notable expansion in the pH profile under acidic conditions. The engineered enzyme demonstrated enhanced activity within the pH range of 3–5 and improved the activity within the pH ranging from 6 to 10. The D31N and D31A substitutions also resulted in a two-unit shift in the pH optimum towards acidic conditions, although this was accompanied by a significant reduction in the enzyme activity. The D31S substitution resulted in a shift in the pH profile towards the alkaline region. The alterations in the enzyme properties observed following the T216K substitution were consistent with the conditions required for the ZHD application as decontamination enzymes at acidic pH values (from 3.0 to 6.0). |
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| issn | 2072-6651 |
| language | English |
| publishDate | 2024-12-01 |
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| series | Toxins |
| spelling | doaj-art-d867a23b471349b49103e2bb6a35d1e32025-08-20T02:01:29ZengMDPI AGToxins2072-66512024-12-01161254010.3390/toxins16120540The Protein Engineering of Zearalenone Hydrolase Results in a Shift in the pH Optimum of the Relative Activity of the EnzymeAnna Dotsenko0Igor Sinelnikov1Ivan Zorov2Yury Denisenko3Aleksandra Rozhkova4Larisa Shcherbakova5Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences (RAS), 119071 Moscow, RussiaFederal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences (RAS), 119071 Moscow, RussiaDepartment of Chemistry, Lomonosov Moscow State University, 119991 Moscow, RussiaFederal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences (RAS), 119071 Moscow, RussiaFederal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences (RAS), 119071 Moscow, RussiaAll-Russian Research Institute of Phytopathology of RAS, Bolshie Vyazemy, 143050 Moscow, RussiaAn acidic shift in the pH profile of <i>Clonostachys rosea</i> zearalenone hydrolase (ZHD), the most effective and well-studied zearalenone-specific lactone hydrolase, is required to extend the range of applications for the enzyme as a decontamination agent in the feed and food production industries. Amino acid substitutions were engineered in the active center of the enzyme to decrease the pKa values of the catalytic residues E126 and H242. The T216K substitution provided a shift in the pH optimum by one unit to the acidic region, accompanied by a notable expansion in the pH profile under acidic conditions. The engineered enzyme demonstrated enhanced activity within the pH range of 3–5 and improved the activity within the pH ranging from 6 to 10. The D31N and D31A substitutions also resulted in a two-unit shift in the pH optimum towards acidic conditions, although this was accompanied by a significant reduction in the enzyme activity. The D31S substitution resulted in a shift in the pH profile towards the alkaline region. The alterations in the enzyme properties observed following the T216K substitution were consistent with the conditions required for the ZHD application as decontamination enzymes at acidic pH values (from 3.0 to 6.0).https://www.mdpi.com/2072-6651/16/12/540zearalenoneenzymatic degradationlactonohydrolaserecombinant proteinsprotein engineeringpH profile |
| spellingShingle | Anna Dotsenko Igor Sinelnikov Ivan Zorov Yury Denisenko Aleksandra Rozhkova Larisa Shcherbakova The Protein Engineering of Zearalenone Hydrolase Results in a Shift in the pH Optimum of the Relative Activity of the Enzyme Toxins zearalenone enzymatic degradation lactonohydrolase recombinant proteins protein engineering pH profile |
| title | The Protein Engineering of Zearalenone Hydrolase Results in a Shift in the pH Optimum of the Relative Activity of the Enzyme |
| title_full | The Protein Engineering of Zearalenone Hydrolase Results in a Shift in the pH Optimum of the Relative Activity of the Enzyme |
| title_fullStr | The Protein Engineering of Zearalenone Hydrolase Results in a Shift in the pH Optimum of the Relative Activity of the Enzyme |
| title_full_unstemmed | The Protein Engineering of Zearalenone Hydrolase Results in a Shift in the pH Optimum of the Relative Activity of the Enzyme |
| title_short | The Protein Engineering of Zearalenone Hydrolase Results in a Shift in the pH Optimum of the Relative Activity of the Enzyme |
| title_sort | protein engineering of zearalenone hydrolase results in a shift in the ph optimum of the relative activity of the enzyme |
| topic | zearalenone enzymatic degradation lactonohydrolase recombinant proteins protein engineering pH profile |
| url | https://www.mdpi.com/2072-6651/16/12/540 |
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