Rationally Guided Improvement of NOV1 Dioxygenase for the Conversion of Lignin-Derived Isoeugenol to Vanillin

Detalhes bibliográficos
Autor(a) principal: De Simone, Mario
Data de Publicação: 2023
Outros Autores: Alvigini, Laura, Alonso-Cotchico, Lur, Brissos, Vânia, Caroli, Jonatan, Lucas, Maria Fátima, Monza, Emanuele, Melo, Eduardo Pinho, Mattevi, Andrea, Martins, Lígia O.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/158531
Resumo: Funding Information: This project has received funding from the Biobased Industries Joint Undertaking (JU) under Grant Agreement 837890 (SMARTBOX) and from Marie Skłodowska-Curie Research and Innovation Staff Exchange (RISE) Grant 824017 (B-Ligzymes). The JU receives support from the European Union’s Horizon 2020 research and innovation program and the Biobased Industries Consortium (BIC). Fundação para a Ciência e a Tecnologia (FCT) additionally supported this work through R&D Unit MOSTMICRO-ITQB (UIDB/04612/2020 and UIDP/04612/2020), LS4FUTURE Associated Laboratory (LA/P/0087/2020), and Project UID/Multi/04326/2019, from the operational programs CRESC Algarve 2020 and COMPETE for 2020 through Project EMBRC.PT ALG-01-0145-FEDER-022121. M.D.S. acknowledges a Ph.D. grant (2020.08246.BD) from FCT. Publisher Copyright: © 2022 American Chemical Society.
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spelling Rationally Guided Improvement of NOV1 Dioxygenase for the Conversion of Lignin-Derived Isoeugenol to VanillinBiochemistryFunding Information: This project has received funding from the Biobased Industries Joint Undertaking (JU) under Grant Agreement 837890 (SMARTBOX) and from Marie Skłodowska-Curie Research and Innovation Staff Exchange (RISE) Grant 824017 (B-Ligzymes). The JU receives support from the European Union’s Horizon 2020 research and innovation program and the Biobased Industries Consortium (BIC). Fundação para a Ciência e a Tecnologia (FCT) additionally supported this work through R&D Unit MOSTMICRO-ITQB (UIDB/04612/2020 and UIDP/04612/2020), LS4FUTURE Associated Laboratory (LA/P/0087/2020), and Project UID/Multi/04326/2019, from the operational programs CRESC Algarve 2020 and COMPETE for 2020 through Project EMBRC.PT ALG-01-0145-FEDER-022121. M.D.S. acknowledges a Ph.D. grant (2020.08246.BD) from FCT. Publisher Copyright: © 2022 American Chemical Society.Biocatalysis is a key tool in both green chemistry and biorefinery fields. NOV1 is a dioxygenase that catalyzes the one-step, coenzyme-free oxidation of isoeugenol into vanillin and holds enormous biotechnological potential for the complete valorization of lignin as a sustainable starting material for biobased chemicals, polymers, and materials. This study integrates computational, kinetic, structural, and biophysical approaches to characterize a new NOV1 variant featuring improved activity and stability compared to those of the wild type. The S283F replacement results in a 2-fold increased turnover rate (kcat) for isoeugenol and a 4-fold higher catalytic efficiency (kcat/Km) for molecular oxygen compared to those of the wild type. Furthermore, the variant exhibits a half-life that is 20-fold higher than that of the wild type, which most likely relates to the enhanced stabilization of the iron cofactor in the active site. Molecular dynamics supports this view, revealing that the S283F replacement decreases the optimal pKa and favors conformations of the iron-coordinating histidines compatible with an increased level of binding to iron. Importantly, whole cells containing the S283F variant catalyze the conversion of ≤100 mM isoeugenol to vanillin, yielding >99% molar conversion yields within 24 h. This integrative strategy provided a new enzyme for biotechnological applications and mechanistic insights that will facilitate the future design of robust and efficient biocatalysts.Instituto de Tecnologia Química e Biológica António Xavier (ITQB)RUNDe Simone, MarioAlvigini, LauraAlonso-Cotchico, LurBrissos, VâniaCaroli, JonatanLucas, Maria FátimaMonza, EmanueleMelo, Eduardo PinhoMattevi, AndreaMartins, Lígia O.2023-09-30T22:19:28Z2023-01-172023-01-17T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article10application/pdfhttp://hdl.handle.net/10362/158531eng0006-2960PURE: 72429487https://doi.org/10.1021/acs.biochem.2c00168info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:41:00Zoai:run.unl.pt:10362/158531Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:57:11.601007Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Rationally Guided Improvement of NOV1 Dioxygenase for the Conversion of Lignin-Derived Isoeugenol to Vanillin
title Rationally Guided Improvement of NOV1 Dioxygenase for the Conversion of Lignin-Derived Isoeugenol to Vanillin
spellingShingle Rationally Guided Improvement of NOV1 Dioxygenase for the Conversion of Lignin-Derived Isoeugenol to Vanillin
De Simone, Mario
Biochemistry
title_short Rationally Guided Improvement of NOV1 Dioxygenase for the Conversion of Lignin-Derived Isoeugenol to Vanillin
title_full Rationally Guided Improvement of NOV1 Dioxygenase for the Conversion of Lignin-Derived Isoeugenol to Vanillin
title_fullStr Rationally Guided Improvement of NOV1 Dioxygenase for the Conversion of Lignin-Derived Isoeugenol to Vanillin
title_full_unstemmed Rationally Guided Improvement of NOV1 Dioxygenase for the Conversion of Lignin-Derived Isoeugenol to Vanillin
title_sort Rationally Guided Improvement of NOV1 Dioxygenase for the Conversion of Lignin-Derived Isoeugenol to Vanillin
author De Simone, Mario
author_facet De Simone, Mario
Alvigini, Laura
Alonso-Cotchico, Lur
Brissos, Vânia
Caroli, Jonatan
Lucas, Maria Fátima
Monza, Emanuele
Melo, Eduardo Pinho
Mattevi, Andrea
Martins, Lígia O.
author_role author
author2 Alvigini, Laura
Alonso-Cotchico, Lur
Brissos, Vânia
Caroli, Jonatan
Lucas, Maria Fátima
Monza, Emanuele
Melo, Eduardo Pinho
Mattevi, Andrea
Martins, Lígia O.
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
RUN
dc.contributor.author.fl_str_mv De Simone, Mario
Alvigini, Laura
Alonso-Cotchico, Lur
Brissos, Vânia
Caroli, Jonatan
Lucas, Maria Fátima
Monza, Emanuele
Melo, Eduardo Pinho
Mattevi, Andrea
Martins, Lígia O.
dc.subject.por.fl_str_mv Biochemistry
topic Biochemistry
description Funding Information: This project has received funding from the Biobased Industries Joint Undertaking (JU) under Grant Agreement 837890 (SMARTBOX) and from Marie Skłodowska-Curie Research and Innovation Staff Exchange (RISE) Grant 824017 (B-Ligzymes). The JU receives support from the European Union’s Horizon 2020 research and innovation program and the Biobased Industries Consortium (BIC). Fundação para a Ciência e a Tecnologia (FCT) additionally supported this work through R&D Unit MOSTMICRO-ITQB (UIDB/04612/2020 and UIDP/04612/2020), LS4FUTURE Associated Laboratory (LA/P/0087/2020), and Project UID/Multi/04326/2019, from the operational programs CRESC Algarve 2020 and COMPETE for 2020 through Project EMBRC.PT ALG-01-0145-FEDER-022121. M.D.S. acknowledges a Ph.D. grant (2020.08246.BD) from FCT. Publisher Copyright: © 2022 American Chemical Society.
publishDate 2023
dc.date.none.fl_str_mv 2023-09-30T22:19:28Z
2023-01-17
2023-01-17T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/158531
url http://hdl.handle.net/10362/158531
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 0006-2960
PURE: 72429487
https://doi.org/10.1021/acs.biochem.2c00168
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 10
application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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