Por favor, use este identificador para citar o enlazar a este item:
http://hdl.handle.net/10261/49136
COMPARTIR / EXPORTAR:
SHARE CORE BASE | |
Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE | |
Título: | Directed evolution of a temperature, peroxide and alkaline pH tolerant versatile peroxidase |
Autor: | García-Ruiz, Eva CSIC ORCID ; González-Pérez, David CSIC ORCID; Ruiz-Dueñas, F. J. CSIC ORCID ; Martínez, Ángel T. CSIC ORCID ; Alcalde Galeote, Miguel CSIC ORCID | Palabras clave: | α-factor prepro-leader Directed evolution Enzyme promiscuity Saccharomyces cerevisiae Versatile peroxidase |
Fecha de publicación: | 7-oct-2011 | Editor: | Portland Press | Citación: | Biochemical Journal 441(1): 487-498(2012) | Resumen: | The VPs (versatile peroxidases) secreted by white-rot fungi are involved in the natural decay of lignin. In the present study, a fusion gene containing the VP from Pleurotus eryngii was subjected to six rounds of directed evolution, achieving a level of secretion in Saccharomyces cerevisiae (21 mg/l) as yet unseen for any ligninolytic peroxidase. The evolved variant for expression harboured four mutations and increased its total VP activity 129-fold. The signal leader processing by the STE13 protease at the Golgi compartment changed as a consequence of overexpression, retaining the additional N-terminal sequence Glu-Ala-Glu-Ala that enhanced secretion. The engineered N-terminally truncated variant displayed similar biochemical properties to those of the non-truncated counterpart in terms of kinetics, stability and spectroscopic features. Additional cycles of evolution raised the T50 8°C and significantly increased the enzyme's stability at alkaline pHs. In addition, the Km for H2O2 was enhanced up to 15-fold while the catalytic efficiency was maintained, and there was an improvement in peroxide stability (with half-lives for H2O2 of 43 min at a H2O2/enzyme molar ratio of 4000:1). Overall, the directed evolution approach described provides a set of strategies for selecting VPs with improvements in secretion, activity and stability | Descripción: | 12 páginas, 5 figuras, 2 tablas -- PAGS nros. 487-498 | Versión del editor: | http://dx.doi.org/10.1042/BJ20111199 | URI: | http://hdl.handle.net/10261/49136 | DOI: | 10.1042/BJ20111199 | ISSN: | 0264-6021 | E-ISSN: | 1470-8728 |
Aparece en las colecciones: | (CIB) Artículos (ICP) Artículos |
Ficheros en este ítem:
Fichero | Descripción | Tamaño | Formato | |
---|---|---|---|---|
Garcia-Ruiz E _al_BiochemJ_441_1_2012.pdf | 1,8 MB | Adobe PDF | Visualizar/Abrir |
CORE Recommender
SCOPUSTM
Citations
94
checked on 24-abr-2024
WEB OF SCIENCETM
Citations
79
checked on 25-feb-2024
Page view(s)
671
checked on 23-abr-2024
Download(s)
741
checked on 23-abr-2024
Google ScholarTM
Check
Altmetric
Altmetric
NOTA: Los ítems de Digital.CSIC están protegidos por copyright, con todos los derechos reservados, a menos que se indique lo contrario.