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Título: | Ancestral Resurrection and Directed Evolution of Fungal Mesozoic Laccases |
Autor: | Gómez-Fernández, Bernardo J.; Risso, Valeria A.; Rueda, Andres; Sánchez-Ruiz, José M.; Alcalde Galeote, Miguel CSIC ORCID | Palabras clave: | Ancestral reconstruction Directed evolution Laccase |
Fecha de publicación: | 2-jul-2020 | Editor: | American Society for Microbiology | Citación: | Applied and Environmental Microbiology 86(14): e00778-20 (2020) | Resumen: | Ancestral sequence reconstruction and resurrection provides useful information for protein engineering, yet its alliance with directed evolution has been little explored. In this study, we have resurrected several ancestral nodes of fungal laccases dating back ∼500 to 250 million years. Unlike modern laccases, the resurrected Mesozoic laccases were readily secreted by yeast, with similar kinetic parameters, a broader stability, and distinct pH activity profiles. The resurrected Agaricomycetes laccase carried 136 ancestral mutations, a molecular testimony to its origin, and it was subjected to directed evolution in order to improve the rate of 1,3-cyclopentanedione oxidation, a β–diketone initiator commonly used in vinyl polymerization reactions. | Versión del editor: | https://doi.org/10.1128/AEM.00778-20 | URI: | http://hdl.handle.net/10261/221368 | DOI: | 10.1128/AEM.00778-20 | ISSN: | 0099-2240 | E-ISSN: | 1098-5336 |
Aparece en las colecciones: | (ICP) Artículos |
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Ancestral Resurrection and Directed Evolution of Fungal.pdf | 4,24 MB | Adobe PDF | Visualizar/Abrir |
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