Por favor, use este identificador para citar o enlazar a este item:
http://hdl.handle.net/10261/22551
COMPARTIR / EXPORTAR:
SHARE CORE BASE | |
Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE | |
Título: | Evolving thermostability in mutant libraries of ligninolytic oxidoreductases expressed in yeast |
Autor: | García-Ruiz, Eva CSIC ORCID ; Maté, Diana M.; Ballesteros Olmo, Antonio CSIC ORCID; Martínez, Ángel T. CSIC ORCID ; Alcalde Galeote, Miguel CSIC ORCID | Fecha de publicación: | 18-mar-2010 | Editor: | BioMed Central | Citación: | Microbial Cell Factories 9:17 (2010) | Resumen: | [Background] In the picture of a laboratory evolution experiment, to improve the thermostability whilst maintaining the activity requires of suitable procedures to generate diversity in combination with robust high-throughput protocols. The current work describes how to achieve this goal by engineering ligninolytic oxidoreductases (a high-redox potential laccase -HRPL- and a versatile peroxidase, -VP-) functionally expressed in Saccharomyces cerevisiae. [Results] Taking advantage of the eukaryotic machinery, complex mutant libraries were constructed by different in vivo recombination approaches and explored for improved stabilities and activities. A reliable high-throughput assay based on the analysis of T50 was employed for discovering thermostable oxidases from mutant libraries in yeast. Both VP and HRPL libraries contained variants with shifts in the T50 values. Stabilizing mutations were found at the surface of the protein establishing new interactions with the surrounding residues. [Conclusions] The existing tradeoff between activity and stability determined from many point mutations discovered by directed evolution and other protein engineering means can be circumvented combining different tools of in vitro evolution. |
Descripción: | 44 pages, 8 figures, 1 table.-- Provisional PDF. | Versión del editor: | http://dx.doi.org/10.1186/1475-2859-9-17 | URI: | http://hdl.handle.net/10261/22551 | DOI: | 10.1186/1475-2859-9-17 | ISSN: | 1475-2859 |
Aparece en las colecciones: | (CIB) Artículos (ICP) Artículos |
Ficheros en este ítem:
Fichero | Descripción | Tamaño | Formato | |
---|---|---|---|---|
1475-2859-9-17.pdf | 2,96 MB | Adobe PDF | Visualizar/Abrir |
CORE Recommender
PubMed Central
Citations
19
checked on 17-mar-2024
SCOPUSTM
Citations
63
checked on 20-mar-2024
WEB OF SCIENCETM
Citations
54
checked on 24-feb-2024
Page view(s)
374
checked on 28-mar-2024
Download(s)
418
checked on 28-mar-2024
Google ScholarTM
Check
Altmetric
Altmetric
Artículos relacionados:
NOTA: Los ítems de Digital.CSIC están protegidos por copyright, con todos los derechos reservados, a menos que se indique lo contrario.