Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives

Autores
Eggerichs, Daniel; Weindorf, Nils; Mascotti, María Laura; Welzel, Natalie; Fraaije, Marco Wilhelmus; Tischler, Dirk
Año de publicación
2023
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Vanillyl alcohol oxidases (VAOs) belong to the 4-phenol oxidases family and are found predominantly in lignin-degrading ascomycetes. Systematical investigation of the enzyme family at the sequence level resulted in discovery and characterization of the second recombinantly produced VAO member, DcVAO, from Diplodia corticola. Remarkably high activities for 2,6-substituted substrates like 4-allyl-2,6-dimethoxy-phenol (3.5 ± 0.02 U mg−1) or 4-(hydroxymethyl)-2,6-dimethoxyphenol (6.3 ± 0.5 U mg−1) were observed, which could be attributed to a Phe to Ala exchange in the catalytic center. In order to rationalize this rare substrate preference among VAOs, we resurrected and characterized three ancestral enzymes and performed mutagenesis analyses. The results indicate that a Cys/Glu exchange was required to retain activity for ɣ-hydroxylations and shifted the acceptance towards benzyl ethers (up to 4.0 ± 0.1 U mg−1). Our findings contribute to the understanding of the functionality of VAO enzyme group, and with DcVAO, we add a new enzyme to the repertoire of ether cleaving biocatalysts.
Fil: Eggerichs, Daniel. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; Alemania
Fil: Weindorf, Nils. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; Alemania
Fil: Mascotti, María Laura. University of Groningen. Faculty of Mathematics and Natural Sciences; Países Bajos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
Fil: Welzel, Natalie. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; Alemania
Fil: Fraaije, Marco Wilhelmus. University of Groningen. Faculty of Mathematics and Natural Sciences; Países Bajos
Fil: Tischler, Dirk. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; Alemania
Materia
ANCESTRAL SEQUENCE RECONSTRUCTION
BIOCATALYSIS
ENZYME ENGINEERING
ETHER CLEAVAGE
FLAVOPROTEIN OXIDASE
LIGNIN
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/227397
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/227397
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivativesEggerichs, DanielWeindorf, NilsMascotti, María LauraWelzel, NatalieFraaije, Marco WilhelmusTischler, DirkANCESTRAL SEQUENCE RECONSTRUCTIONBIOCATALYSISENZYME ENGINEERINGETHER CLEAVAGEFLAVOPROTEIN OXIDASELIGNINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Vanillyl alcohol oxidases (VAOs) belong to the 4-phenol oxidases family and are found predominantly in lignin-degrading ascomycetes. Systematical investigation of the enzyme family at the sequence level resulted in discovery and characterization of the second recombinantly produced VAO member, DcVAO, from Diplodia corticola. Remarkably high activities for 2,6-substituted substrates like 4-allyl-2,6-dimethoxy-phenol (3.5 ± 0.02 U mg−1) or 4-(hydroxymethyl)-2,6-dimethoxyphenol (6.3 ± 0.5 U mg−1) were observed, which could be attributed to a Phe to Ala exchange in the catalytic center. In order to rationalize this rare substrate preference among VAOs, we resurrected and characterized three ancestral enzymes and performed mutagenesis analyses. The results indicate that a Cys/Glu exchange was required to retain activity for ɣ-hydroxylations and shifted the acceptance towards benzyl ethers (up to 4.0 ± 0.1 U mg−1). Our findings contribute to the understanding of the functionality of VAO enzyme group, and with DcVAO, we add a new enzyme to the repertoire of ether cleaving biocatalysts.Fil: Eggerichs, Daniel. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; AlemaniaFil: Weindorf, Nils. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; AlemaniaFil: Mascotti, María Laura. University of Groningen. Faculty of Mathematics and Natural Sciences; Países Bajos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; ArgentinaFil: Welzel, Natalie. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; AlemaniaFil: Fraaije, Marco Wilhelmus. University of Groningen. Faculty of Mathematics and Natural Sciences; Países BajosFil: Tischler, Dirk. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; AlemaniaAmerican Society for Biochemistry and Molecular Biology2023-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/227397Eggerichs, Daniel; Weindorf, Nils; Mascotti, María Laura; Welzel, Natalie; Fraaije, Marco Wilhelmus; et al.; Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 299; 7; 6-2023; 1-290021-9258CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbc.2023.104898info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0021925823019269info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:18:23Zoai:ri.conicet.gov.ar:11336/227397instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 11:18:24.036CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives
title Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives
spellingShingle Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives
Eggerichs, Daniel
ANCESTRAL SEQUENCE RECONSTRUCTION
BIOCATALYSIS
ENZYME ENGINEERING
ETHER CLEAVAGE
FLAVOPROTEIN OXIDASE
LIGNIN
title_short Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives
title_full Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives
title_fullStr Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives
title_full_unstemmed Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives
title_sort Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives
dc.creator.none.fl_str_mv Eggerichs, Daniel
Weindorf, Nils
Mascotti, María Laura
Welzel, Natalie
Fraaije, Marco Wilhelmus
Tischler, Dirk
author Eggerichs, Daniel
author_facet Eggerichs, Daniel
Weindorf, Nils
Mascotti, María Laura
Welzel, Natalie
Fraaije, Marco Wilhelmus
Tischler, Dirk
author_role author
author2 Weindorf, Nils
Mascotti, María Laura
Welzel, Natalie
Fraaije, Marco Wilhelmus
Tischler, Dirk
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv ANCESTRAL SEQUENCE RECONSTRUCTION
BIOCATALYSIS
ENZYME ENGINEERING
ETHER CLEAVAGE
FLAVOPROTEIN OXIDASE
LIGNIN
topic ANCESTRAL SEQUENCE RECONSTRUCTION
BIOCATALYSIS
ENZYME ENGINEERING
ETHER CLEAVAGE
FLAVOPROTEIN OXIDASE
LIGNIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Vanillyl alcohol oxidases (VAOs) belong to the 4-phenol oxidases family and are found predominantly in lignin-degrading ascomycetes. Systematical investigation of the enzyme family at the sequence level resulted in discovery and characterization of the second recombinantly produced VAO member, DcVAO, from Diplodia corticola. Remarkably high activities for 2,6-substituted substrates like 4-allyl-2,6-dimethoxy-phenol (3.5 ± 0.02 U mg−1) or 4-(hydroxymethyl)-2,6-dimethoxyphenol (6.3 ± 0.5 U mg−1) were observed, which could be attributed to a Phe to Ala exchange in the catalytic center. In order to rationalize this rare substrate preference among VAOs, we resurrected and characterized three ancestral enzymes and performed mutagenesis analyses. The results indicate that a Cys/Glu exchange was required to retain activity for ɣ-hydroxylations and shifted the acceptance towards benzyl ethers (up to 4.0 ± 0.1 U mg−1). Our findings contribute to the understanding of the functionality of VAO enzyme group, and with DcVAO, we add a new enzyme to the repertoire of ether cleaving biocatalysts.
Fil: Eggerichs, Daniel. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; Alemania
Fil: Weindorf, Nils. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; Alemania
Fil: Mascotti, María Laura. University of Groningen. Faculty of Mathematics and Natural Sciences; Países Bajos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Instituto Multidisciplinario de Investigaciones Biológicas de San Luis; Argentina
Fil: Welzel, Natalie. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; Alemania
Fil: Fraaije, Marco Wilhelmus. University of Groningen. Faculty of Mathematics and Natural Sciences; Países Bajos
Fil: Tischler, Dirk. Ruhr-universität Bochum. Fakultät Für Biologie & Biotechnologie; Alemania
description Vanillyl alcohol oxidases (VAOs) belong to the 4-phenol oxidases family and are found predominantly in lignin-degrading ascomycetes. Systematical investigation of the enzyme family at the sequence level resulted in discovery and characterization of the second recombinantly produced VAO member, DcVAO, from Diplodia corticola. Remarkably high activities for 2,6-substituted substrates like 4-allyl-2,6-dimethoxy-phenol (3.5 ± 0.02 U mg−1) or 4-(hydroxymethyl)-2,6-dimethoxyphenol (6.3 ± 0.5 U mg−1) were observed, which could be attributed to a Phe to Ala exchange in the catalytic center. In order to rationalize this rare substrate preference among VAOs, we resurrected and characterized three ancestral enzymes and performed mutagenesis analyses. The results indicate that a Cys/Glu exchange was required to retain activity for ɣ-hydroxylations and shifted the acceptance towards benzyl ethers (up to 4.0 ± 0.1 U mg−1). Our findings contribute to the understanding of the functionality of VAO enzyme group, and with DcVAO, we add a new enzyme to the repertoire of ether cleaving biocatalysts.
publishDate 2023
dc.date.none.fl_str_mv 2023-06
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/227397
Eggerichs, Daniel; Weindorf, Nils; Mascotti, María Laura; Welzel, Natalie; Fraaije, Marco Wilhelmus; et al.; Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 299; 7; 6-2023; 1-29
0021-9258
CONICET Digital
CONICET
url http://hdl.handle.net/11336/227397
identifier_str_mv Eggerichs, Daniel; Weindorf, Nils; Mascotti, María Laura; Welzel, Natalie; Fraaije, Marco Wilhelmus; et al.; Vanillyl alcohol oxidase from Diplodia corticola: Residues Ala420 and Glu466 allow for efficient catalysis of syringyl derivatives; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 299; 7; 6-2023; 1-29
0021-9258
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbc.2023.104898
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0021925823019269
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.982451

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