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Directed evolution unlocks oxygen reactivity for a nicotine-degrading flavoenzyme
The flavoenzyme nicotine oxidoreductase (NicA2) is a promising injectable treatment to aid in the cessation of smoking, a behavior responsible for one in ten deaths worldwide. NicA2 acts by degrading nicotine in the bloodstream before it reaches the brain. Clinical use of NicA2 is limited by its poo...
Autores principales: | , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group US
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10611581/ https://www.ncbi.nlm.nih.gov/pubmed/37770699 http://dx.doi.org/10.1038/s41589-023-01426-y |
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author | Dulchavsky, Mark Mitra, Rishav Wu, Kevin Li, Joshua Boer, Karli Liu, Xiaomeng Zhang, Zhiyao Vasquez, Cristian Clark, Christopher T. Funckes, Kaitrin Shankar, Kokila Bonnet-Zahedi, Selene Siddiq, Mohammad Sepulveda, Yadira Suhandynata, Raymond T. Momper, Jeremiah D. Calabrese, Antonio N. George, Olivier Stull, Frederick Bardwell, James C. A. |
author_facet | Dulchavsky, Mark Mitra, Rishav Wu, Kevin Li, Joshua Boer, Karli Liu, Xiaomeng Zhang, Zhiyao Vasquez, Cristian Clark, Christopher T. Funckes, Kaitrin Shankar, Kokila Bonnet-Zahedi, Selene Siddiq, Mohammad Sepulveda, Yadira Suhandynata, Raymond T. Momper, Jeremiah D. Calabrese, Antonio N. George, Olivier Stull, Frederick Bardwell, James C. A. |
author_sort | Dulchavsky, Mark |
collection | PubMed |
description | The flavoenzyme nicotine oxidoreductase (NicA2) is a promising injectable treatment to aid in the cessation of smoking, a behavior responsible for one in ten deaths worldwide. NicA2 acts by degrading nicotine in the bloodstream before it reaches the brain. Clinical use of NicA2 is limited by its poor catalytic activity in the absence of its natural electron acceptor CycN. Without CycN, NicA2 is instead oxidized slowly by dioxygen (O(2)), necessitating unfeasibly large doses in a therapeutic setting. Here, we report a genetic selection strategy that directly links CycN-independent activity of NicA2 to growth of Pseudomonas putida S16. This selection enabled us to evolve NicA2 variants with substantial improvement in their rate of oxidation by O(2). The encoded mutations cluster around a putative O(2) tunnel, increasing flexibility and accessibility to O(2) in this region. These mutations further confer desirable clinical properties. A variant form of NicA2 is tenfold more effective than the wild type at degrading nicotine in the bloodstream of rats. [Image: see text] |
format | Online Article Text |
id | pubmed-10611581 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group US |
record_format | MEDLINE/PubMed |
spelling | pubmed-106115812023-10-29 Directed evolution unlocks oxygen reactivity for a nicotine-degrading flavoenzyme Dulchavsky, Mark Mitra, Rishav Wu, Kevin Li, Joshua Boer, Karli Liu, Xiaomeng Zhang, Zhiyao Vasquez, Cristian Clark, Christopher T. Funckes, Kaitrin Shankar, Kokila Bonnet-Zahedi, Selene Siddiq, Mohammad Sepulveda, Yadira Suhandynata, Raymond T. Momper, Jeremiah D. Calabrese, Antonio N. George, Olivier Stull, Frederick Bardwell, James C. A. Nat Chem Biol Article The flavoenzyme nicotine oxidoreductase (NicA2) is a promising injectable treatment to aid in the cessation of smoking, a behavior responsible for one in ten deaths worldwide. NicA2 acts by degrading nicotine in the bloodstream before it reaches the brain. Clinical use of NicA2 is limited by its poor catalytic activity in the absence of its natural electron acceptor CycN. Without CycN, NicA2 is instead oxidized slowly by dioxygen (O(2)), necessitating unfeasibly large doses in a therapeutic setting. Here, we report a genetic selection strategy that directly links CycN-independent activity of NicA2 to growth of Pseudomonas putida S16. This selection enabled us to evolve NicA2 variants with substantial improvement in their rate of oxidation by O(2). The encoded mutations cluster around a putative O(2) tunnel, increasing flexibility and accessibility to O(2) in this region. These mutations further confer desirable clinical properties. A variant form of NicA2 is tenfold more effective than the wild type at degrading nicotine in the bloodstream of rats. [Image: see text] Nature Publishing Group US 2023-09-28 2023 /pmc/articles/PMC10611581/ /pubmed/37770699 http://dx.doi.org/10.1038/s41589-023-01426-y Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Dulchavsky, Mark Mitra, Rishav Wu, Kevin Li, Joshua Boer, Karli Liu, Xiaomeng Zhang, Zhiyao Vasquez, Cristian Clark, Christopher T. Funckes, Kaitrin Shankar, Kokila Bonnet-Zahedi, Selene Siddiq, Mohammad Sepulveda, Yadira Suhandynata, Raymond T. Momper, Jeremiah D. Calabrese, Antonio N. George, Olivier Stull, Frederick Bardwell, James C. A. Directed evolution unlocks oxygen reactivity for a nicotine-degrading flavoenzyme |
title | Directed evolution unlocks oxygen reactivity for a nicotine-degrading flavoenzyme |
title_full | Directed evolution unlocks oxygen reactivity for a nicotine-degrading flavoenzyme |
title_fullStr | Directed evolution unlocks oxygen reactivity for a nicotine-degrading flavoenzyme |
title_full_unstemmed | Directed evolution unlocks oxygen reactivity for a nicotine-degrading flavoenzyme |
title_short | Directed evolution unlocks oxygen reactivity for a nicotine-degrading flavoenzyme |
title_sort | directed evolution unlocks oxygen reactivity for a nicotine-degrading flavoenzyme |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10611581/ https://www.ncbi.nlm.nih.gov/pubmed/37770699 http://dx.doi.org/10.1038/s41589-023-01426-y |
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