Development of a flow microreactor with immobilized catechol-1,2-dioxygenase for the biocatalytic production of cis,cis-muconic acid

A low cost microreactor was developed by stable immobilization of catechol-1,2-dioxygenase for the flow biocatalysis of catechol to produce cis,cis-muconic acid, a direct precursor of industrially relevant adipic acid. The microreactors of polydimethylsiloxane were manufactured with different design...

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Autores: Rosso, Samuele, Almazán, Fernando, Pellejero, Ismael, Gandía Pascual, Luis, Dezzani, Sabrina, Barbieri, Lisa, Gilardi, Gianfranco, Valetti, Francesca, Di Nardo, Giovanna
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2025
País:España
Institución:Universidad Pública de Navarra
Repositorio:Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
OAI Identifier:oai:academica-e.unavarra.es:2454/55618
Acceso en línea:https://hdl.handle.net/2454/55618
Access Level:acceso abierto
Palabra clave:Flow biocatalysis
PDMS microreactors
cis,cis-muconic acid
Enzyme immobilization
Catechol-1,2-dioxygenase
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spelling Development of a flow microreactor with immobilized catechol-1,2-dioxygenase for the biocatalytic production of cis,cis-muconic acidRosso, SamueleAlmazán, FernandoPellejero, IsmaelGandía Pascual, LuisDezzani, SabrinaBarbieri, LisaGilardi, GianfrancoValetti, FrancescaDi Nardo, GiovannaFlow biocatalysisPDMS microreactorscis,cis-muconic acidEnzyme immobilizationCatechol-1,2-dioxygenaseA low cost microreactor was developed by stable immobilization of catechol-1,2-dioxygenase for the flow biocatalysis of catechol to produce cis,cis-muconic acid, a direct precursor of industrially relevant adipic acid. The microreactors of polydimethylsiloxane were manufactured with different designs by straightforward, cost- effective soft lithography techniques, and were tested with different immobilization strategies focused on preserving enzyme catalytic activity and stability over time. The best immobilization strategy was covalent immobilization by amino-silanization followed by glutaraldehyde linker grafting. The biocatalytic activity of the immobilized enzyme was assessed by measuring muconic acid production through the flow of 50 µM catechol at room temperature. Catechol conversion rates of up to 95 % were achieved over a 5-hour period, and volumes of up to 180 mL were processed using only 0.32 nmol of immobilized protein, resulting in the production of 1.33 mg⋅L 1⋅h 1. This work demonstrates the feasibility of using microreactor-based flow biocatalysis for sustainable muconic acid production and highlights its promising application at industrial scale.Financial support from University of Turin “GRANT for INTERNATIONALIZATION - GFI 2022” and the Spanish Ministerio de Ciencia e Innovación and Spanish Agencia Estatal de Investigación (grants PID2019–106687RJ-I00, PID2021–127265OB-C21) is gratefully acknowledged. Open access funding provided by University of Torino.ElsevierCienciasZientziakInstitute for Advanced Materials and Mathematics - INAMAT22025info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/mswordhttps://hdl.handle.net/2454/55618reponame:Academica-e. Repositorio Institucional de la Universidad Pública de Navarrainstname:Universidad Pública de NavarraInglésinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-106687RJ-I00info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-127265OB-C21© 2025 The Author(s). This is an open access article under the CC BY license.https://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:academica-e.unavarra.es:2454/556182026-06-17T12:41:47Z
dc.title.none.fl_str_mv Development of a flow microreactor with immobilized catechol-1,2-dioxygenase for the biocatalytic production of cis,cis-muconic acid
title Development of a flow microreactor with immobilized catechol-1,2-dioxygenase for the biocatalytic production of cis,cis-muconic acid
spellingShingle Development of a flow microreactor with immobilized catechol-1,2-dioxygenase for the biocatalytic production of cis,cis-muconic acid
Rosso, Samuele
Flow biocatalysis
PDMS microreactors
cis,cis-muconic acid
Enzyme immobilization
Catechol-1,2-dioxygenase
title_short Development of a flow microreactor with immobilized catechol-1,2-dioxygenase for the biocatalytic production of cis,cis-muconic acid
title_full Development of a flow microreactor with immobilized catechol-1,2-dioxygenase for the biocatalytic production of cis,cis-muconic acid
title_fullStr Development of a flow microreactor with immobilized catechol-1,2-dioxygenase for the biocatalytic production of cis,cis-muconic acid
title_full_unstemmed Development of a flow microreactor with immobilized catechol-1,2-dioxygenase for the biocatalytic production of cis,cis-muconic acid
title_sort Development of a flow microreactor with immobilized catechol-1,2-dioxygenase for the biocatalytic production of cis,cis-muconic acid
dc.creator.none.fl_str_mv Rosso, Samuele
Almazán, Fernando
Pellejero, Ismael
Gandía Pascual, Luis
Dezzani, Sabrina
Barbieri, Lisa
Gilardi, Gianfranco
Valetti, Francesca
Di Nardo, Giovanna
author Rosso, Samuele
author_facet Rosso, Samuele
Almazán, Fernando
Pellejero, Ismael
Gandía Pascual, Luis
Dezzani, Sabrina
Barbieri, Lisa
Gilardi, Gianfranco
Valetti, Francesca
Di Nardo, Giovanna
author_role author
author2 Almazán, Fernando
Pellejero, Ismael
Gandía Pascual, Luis
Dezzani, Sabrina
Barbieri, Lisa
Gilardi, Gianfranco
Valetti, Francesca
Di Nardo, Giovanna
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ciencias
Zientziak
Institute for Advanced Materials and Mathematics - INAMAT2
dc.subject.none.fl_str_mv Flow biocatalysis
PDMS microreactors
cis,cis-muconic acid
Enzyme immobilization
Catechol-1,2-dioxygenase
topic Flow biocatalysis
PDMS microreactors
cis,cis-muconic acid
Enzyme immobilization
Catechol-1,2-dioxygenase
description A low cost microreactor was developed by stable immobilization of catechol-1,2-dioxygenase for the flow biocatalysis of catechol to produce cis,cis-muconic acid, a direct precursor of industrially relevant adipic acid. The microreactors of polydimethylsiloxane were manufactured with different designs by straightforward, cost- effective soft lithography techniques, and were tested with different immobilization strategies focused on preserving enzyme catalytic activity and stability over time. The best immobilization strategy was covalent immobilization by amino-silanization followed by glutaraldehyde linker grafting. The biocatalytic activity of the immobilized enzyme was assessed by measuring muconic acid production through the flow of 50 µM catechol at room temperature. Catechol conversion rates of up to 95 % were achieved over a 5-hour period, and volumes of up to 180 mL were processed using only 0.32 nmol of immobilized protein, resulting in the production of 1.33 mg⋅L 1⋅h 1. This work demonstrates the feasibility of using microreactor-based flow biocatalysis for sustainable muconic acid production and highlights its promising application at industrial scale.
publishDate 2025
dc.date.none.fl_str_mv 2025
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2454/55618
url https://hdl.handle.net/2454/55618
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-106687RJ-I00
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-127265OB-C21
dc.rights.none.fl_str_mv © 2025 The Author(s). This is an open access article under the CC BY license.
https://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv © 2025 The Author(s). This is an open access article under the CC BY license.
https://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/msword
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
instname:Universidad Pública de Navarra
instname_str Universidad Pública de Navarra
reponame_str Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
collection Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
repository.name.fl_str_mv
repository.mail.fl_str_mv
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