Colloidal Protein–Silver Nanoparticle Metalloenzyme as Artificial Redox Biocatalyst

Efficient and sustainable catalytic processes are crucial for advancing green chemical manufacturing. Here, we describe the synthesis of novel silver artificial metalloenzymes in colloidal form in aqueous media and room temperature. The strategy is based on the in situ generation of silver nanoparti...

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Autores: Bojanov, Glenn, García-Sanz, Carla, Palomo, José Miguel
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2025
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/379479
Acesso em linha:http://hdl.handle.net/10261/379479
Access Level:acceso abierto
Palavra-chave:Silver
Metalloenzymes
Redox enzymes
Colloidal
Nanoparticles
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spelling Colloidal Protein–Silver Nanoparticle Metalloenzyme as Artificial Redox BiocatalystBojanov, GlennGarcía-Sanz, CarlaPalomo, José MiguelSilverMetalloenzymesRedox enzymesColloidalNanoparticlesEfficient and sustainable catalytic processes are crucial for advancing green chemical manufacturing. Here, we describe the synthesis of novel silver artificial metalloenzymes in colloidal form in aqueous media and room temperature. The strategy is based on the in situ generation of silver nanoparticles by a genetically modified Geobacillus thermocatenulatus lipase (GTL) in the active site as an inducer and scaffold protein, producing an enzyme–Ag bioconjugate. Using a structural analysis of the formation of silver nanoparticles by XRD and UV spectra, we found the formation of Ag2O species with nanoparticles of around 11 nm average diameter size. Gel filtration chromatography demonstrated the presence of single protein molecules in the bioconjugates, although silver nanoparticles were initially formed by cysteine coordination in the active site but later were formed in other parts of the protein (five AgNPs per molecules, which is in concordance with the UV size). The enzyme structure was altered after nanoparticle formation and Ag-S interaction, which was observed in fluorescence analysis. This new enzyme showed reductive activity against p-nitrophenol to p-amino and a high conversion > 99% in the reduction of acetophenone to phenylethanol, although the enantioselective was quite moderate but higher in water that in the presence of co-solvents. Finally, oxidase-like activity was evaluated in the direct oxidation of phenylethanol to acetophenone in water, obtained at around a 23% yield of ketone after 60 h.The authors thank the funding support from the Spanish National Research Council (CSIC) and the European Commission for funding the project, grant no. 101060130, HORIZON-WIDERA-2021-ACCESS-02-01.Peer reviewedMultidisciplinary Digital Publishing InstituteEuropean CommissionConsejo Superior de Investigaciones Científicas (España)Palomo, José Miguel [0000-0002-6464-1216]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2025202520252025info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/379479reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/EC/HE/101060130The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.3390/catal15010061https://doi.org/10.3390/catal15010061Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3794792026-05-22T06:33:51Z
dc.title.none.fl_str_mv Colloidal Protein–Silver Nanoparticle Metalloenzyme as Artificial Redox Biocatalyst
title Colloidal Protein–Silver Nanoparticle Metalloenzyme as Artificial Redox Biocatalyst
spellingShingle Colloidal Protein–Silver Nanoparticle Metalloenzyme as Artificial Redox Biocatalyst
Bojanov, Glenn
Silver
Metalloenzymes
Redox enzymes
Colloidal
Nanoparticles
title_short Colloidal Protein–Silver Nanoparticle Metalloenzyme as Artificial Redox Biocatalyst
title_full Colloidal Protein–Silver Nanoparticle Metalloenzyme as Artificial Redox Biocatalyst
title_fullStr Colloidal Protein–Silver Nanoparticle Metalloenzyme as Artificial Redox Biocatalyst
title_full_unstemmed Colloidal Protein–Silver Nanoparticle Metalloenzyme as Artificial Redox Biocatalyst
title_sort Colloidal Protein–Silver Nanoparticle Metalloenzyme as Artificial Redox Biocatalyst
dc.creator.none.fl_str_mv Bojanov, Glenn
García-Sanz, Carla
Palomo, José Miguel
author Bojanov, Glenn
author_facet Bojanov, Glenn
García-Sanz, Carla
Palomo, José Miguel
author_role author
author2 García-Sanz, Carla
Palomo, José Miguel
author2_role author
author
dc.contributor.none.fl_str_mv European Commission
Consejo Superior de Investigaciones Científicas (España)
Palomo, José Miguel [0000-0002-6464-1216]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Silver
Metalloenzymes
Redox enzymes
Colloidal
Nanoparticles
topic Silver
Metalloenzymes
Redox enzymes
Colloidal
Nanoparticles
description Efficient and sustainable catalytic processes are crucial for advancing green chemical manufacturing. Here, we describe the synthesis of novel silver artificial metalloenzymes in colloidal form in aqueous media and room temperature. The strategy is based on the in situ generation of silver nanoparticles by a genetically modified Geobacillus thermocatenulatus lipase (GTL) in the active site as an inducer and scaffold protein, producing an enzyme–Ag bioconjugate. Using a structural analysis of the formation of silver nanoparticles by XRD and UV spectra, we found the formation of Ag2O species with nanoparticles of around 11 nm average diameter size. Gel filtration chromatography demonstrated the presence of single protein molecules in the bioconjugates, although silver nanoparticles were initially formed by cysteine coordination in the active site but later were formed in other parts of the protein (five AgNPs per molecules, which is in concordance with the UV size). The enzyme structure was altered after nanoparticle formation and Ag-S interaction, which was observed in fluorescence analysis. This new enzyme showed reductive activity against p-nitrophenol to p-amino and a high conversion > 99% in the reduction of acetophenone to phenylethanol, although the enantioselective was quite moderate but higher in water that in the presence of co-solvents. Finally, oxidase-like activity was evaluated in the direct oxidation of phenylethanol to acetophenone in water, obtained at around a 23% yield of ketone after 60 h.
publishDate 2025
dc.date.none.fl_str_mv 2025
2025
2025
2025
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/379479
url http://hdl.handle.net/10261/379479
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/EC/HE/101060130
The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.3390/catal15010061
https://doi.org/10.3390/catal15010061
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.812429

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