In vitro Production of Hemin-Based Artificial Metalloenzymes
dc.contributor.author | López Domene, Rocío | |
dc.contributor.author | Manteca González, Aitor | |
dc.contributor.author | Rodríguez Abetxuko, Andoni | |
dc.contributor.author | Beloqui Elizazu, Ana | |
dc.contributor.author | López Cortajarena, Aitziber | |
dc.date.accessioned | 2024-04-15T15:48:50Z | |
dc.date.available | 2024-04-15T15:48:50Z | |
dc.date.issued | 2024-02 | |
dc.identifier.citation | Chemistry - A European Journal 30(11) : (2024) // Article ID e202303254 | es_ES |
dc.identifier.issn | 1521-3765 | |
dc.identifier.issn | 0947-6539 | |
dc.identifier.uri | http://hdl.handle.net/10810/66688 | |
dc.description.abstract | Developing enzyme alternatives is pivotal to improving and enabling new processes in biotechnology and industry. Artificial metalloenzymes (ArMs) are combinations of protein scaffolds with metal elements, such as metal nanoclusters or metal-containing molecules with specific catalytic properties, which can be customized. Here, we engineered an ArM based on the consensus tetratricopeptide repeat (CTPR) scaffold by introducing a unique histidine residue to coordinate the hemin cofactor. Our results show that this engineered system exhibits robust peroxidase-like catalytic activity driven by the hemin. The expression of the scaffold and subsequent coordination of hemin was achieved by recombinant expression in bulk and through in vitro transcription and translation systems in water-in-oil drops. The ability to synthesize this system in emulsio paves the way to improve its properties by means of droplet microfluidic screenings, facilitating the exploration of the protein combinatorial space to discover improved or novel catalytic activities. | es_ES |
dc.description.sponsorship | A.M. is financed by grant 2022-FELL-000011-01 funded by Gipuzkoa Fellows Program (Diputación Foral de Gipuzkoa) and grant “EPINPOC” co-funded by AECT Euroregion New Aquitaine-Navarra-Basque Country. A.L.C. acknowledges financial support by the Agencia Estatal de Investigación, Grants: PID2019-111649RB−I00 and PID2022-137977OB-I00 funded by MCIN/AEI/ 10.13039/501100011033 and Grant PDC2021-120957-I00 funded by MCIN/AEI/ 10.13039/501100011033 and by the “European Union NextGenerationEU/PRTR”. A.L.C. also acknowledges financial support from Diputación Foral de Gipuzkoa grant 2023-QUAN-000023-01. This work was performed under the Maria de Maeztu Units of Excellence Program from Q5 the Spanish State Research Agency grant no. MDM-2017-0720. A.B. gratefully acknowledges the financial support from the Spanish Research Agency (AEI) for the financial support (PID2019-110239RB−I00 and PID2022-142128NB-I00 funded by MCIN/ AEI/10.13039/501100011033/ and by the ”European Union NextGenerationEU/PRTR”; RYC2018-025923-I from RyC program - MCIN/ AEI /10.13039/501100011033 and FSE “invierte en tu futuro”), BBVA Foundation - IN[21]_CBB_QUI_0086, and UPV/EHU- GIU21-033). We thank Thomas Beneyton and J−C Baret for their relentless support with the device fabrication and the mounting of the optical setup. | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | Wiley | es_ES |
dc.relation | info:eu-repo/grantAgreement/MICIU/RYC2018-025923-I | es_ES |
dc.relation | info:eu-repo/grantAgreement/MICIU/PID2019-110239RB−I00 | es_ES |
dc.relation | info:eu-repo/grantAgreement/MICINN/PID2022-142128NB-I00 | es_ES |
dc.relation | info:eu-repo/grantAgreement/MICINN/PDC2021-120957-I00 | es_ES |
dc.relation | info:eu-repo/grantAgreement/MICIU/PID2019-111649RB−I00 | es_ES |
dc.rights | info:eu-repo/semantics/openAccess | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by-nc/3.0/es/ | * |
dc.subject | protein design | es_ES |
dc.subject | repeat proteins | es_ES |
dc.subject | metalloenzymes | es_ES |
dc.subject | hemin | es_ES |
dc.subject | microfluidics | es_ES |
dc.subject | peroxidase | es_ES |
dc.subject | synthetic biology | es_ES |
dc.title | In vitro Production of Hemin-Based Artificial Metalloenzymes | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.rights.holder | © 2023 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. | es_ES |
dc.rights.holder | Atribución-NoComercial 3.0 España | * |
dc.relation.publisherversion | https://chemistry-europe.onlinelibrary.wiley.com/doi/full/10.1002/chem.202303254 | es_ES |
dc.identifier.doi | 10.1002/chem.202303254 | |
dc.departamentoes | Química aplicada | es_ES |
dc.departamentoeu | Kimika aplikatua | es_ES |
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This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.