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Directed -in vitro- evolution of Precambrian and extant Rubiscos

Rubisco is an ancient, catalytically conserved yet slow enzyme, which plays a central role in the biosphere’s carbon cycle. The design of Rubiscos to increase agricultural productivity has hitherto relied on the use of in vivo selection systems, precluding the exploration of biochemical traits that...

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Autores principales: Gomez-Fernandez, Bernardo J., Garcia-Ruiz, Eva, Martin-Diaz, Javier, Gomez de Santos, Patricia, Santos-Moriano, Paloma, Plou, Francisco J., Ballesteros, Antonio, Garcia, Monica, Rodriguez, Marisa, Risso, Valeria A., Sanchez-Ruiz, Jose M., Whitney, Spencer M., Alcalde, Miguel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5883036/
https://www.ncbi.nlm.nih.gov/pubmed/29615759
http://dx.doi.org/10.1038/s41598-018-23869-3
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author Gomez-Fernandez, Bernardo J.
Garcia-Ruiz, Eva
Martin-Diaz, Javier
Gomez de Santos, Patricia
Santos-Moriano, Paloma
Plou, Francisco J.
Ballesteros, Antonio
Garcia, Monica
Rodriguez, Marisa
Risso, Valeria A.
Sanchez-Ruiz, Jose M.
Whitney, Spencer M.
Alcalde, Miguel
author_facet Gomez-Fernandez, Bernardo J.
Garcia-Ruiz, Eva
Martin-Diaz, Javier
Gomez de Santos, Patricia
Santos-Moriano, Paloma
Plou, Francisco J.
Ballesteros, Antonio
Garcia, Monica
Rodriguez, Marisa
Risso, Valeria A.
Sanchez-Ruiz, Jose M.
Whitney, Spencer M.
Alcalde, Miguel
author_sort Gomez-Fernandez, Bernardo J.
collection PubMed
description Rubisco is an ancient, catalytically conserved yet slow enzyme, which plays a central role in the biosphere’s carbon cycle. The design of Rubiscos to increase agricultural productivity has hitherto relied on the use of in vivo selection systems, precluding the exploration of biochemical traits that are not wired to cell survival. We present a directed -in vitro- evolution platform that extracts the enzyme from its biological context to provide a new avenue for Rubisco engineering. Precambrian and extant form II Rubiscos were subjected to an ensemble of directed evolution strategies aimed at improving thermostability. The most recent ancestor of proteobacteria -dating back 2.4 billion years- was uniquely tolerant to mutagenic loading. Adaptive evolution, focused evolution and genetic drift revealed a panel of thermostable mutants, some deviating from the characteristic trade-offs in CO(2)-fixing speed and specificity. Our findings provide a novel approach for identifying Rubisco variants with improved catalytic evolution potential.
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spelling pubmed-58830362018-04-09 Directed -in vitro- evolution of Precambrian and extant Rubiscos Gomez-Fernandez, Bernardo J. Garcia-Ruiz, Eva Martin-Diaz, Javier Gomez de Santos, Patricia Santos-Moriano, Paloma Plou, Francisco J. Ballesteros, Antonio Garcia, Monica Rodriguez, Marisa Risso, Valeria A. Sanchez-Ruiz, Jose M. Whitney, Spencer M. Alcalde, Miguel Sci Rep Article Rubisco is an ancient, catalytically conserved yet slow enzyme, which plays a central role in the biosphere’s carbon cycle. The design of Rubiscos to increase agricultural productivity has hitherto relied on the use of in vivo selection systems, precluding the exploration of biochemical traits that are not wired to cell survival. We present a directed -in vitro- evolution platform that extracts the enzyme from its biological context to provide a new avenue for Rubisco engineering. Precambrian and extant form II Rubiscos were subjected to an ensemble of directed evolution strategies aimed at improving thermostability. The most recent ancestor of proteobacteria -dating back 2.4 billion years- was uniquely tolerant to mutagenic loading. Adaptive evolution, focused evolution and genetic drift revealed a panel of thermostable mutants, some deviating from the characteristic trade-offs in CO(2)-fixing speed and specificity. Our findings provide a novel approach for identifying Rubisco variants with improved catalytic evolution potential. Nature Publishing Group UK 2018-04-03 /pmc/articles/PMC5883036/ /pubmed/29615759 http://dx.doi.org/10.1038/s41598-018-23869-3 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Gomez-Fernandez, Bernardo J.
Garcia-Ruiz, Eva
Martin-Diaz, Javier
Gomez de Santos, Patricia
Santos-Moriano, Paloma
Plou, Francisco J.
Ballesteros, Antonio
Garcia, Monica
Rodriguez, Marisa
Risso, Valeria A.
Sanchez-Ruiz, Jose M.
Whitney, Spencer M.
Alcalde, Miguel
Directed -in vitro- evolution of Precambrian and extant Rubiscos
title Directed -in vitro- evolution of Precambrian and extant Rubiscos
title_full Directed -in vitro- evolution of Precambrian and extant Rubiscos
title_fullStr Directed -in vitro- evolution of Precambrian and extant Rubiscos
title_full_unstemmed Directed -in vitro- evolution of Precambrian and extant Rubiscos
title_short Directed -in vitro- evolution of Precambrian and extant Rubiscos
title_sort directed -in vitro- evolution of precambrian and extant rubiscos
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5883036/
https://www.ncbi.nlm.nih.gov/pubmed/29615759
http://dx.doi.org/10.1038/s41598-018-23869-3
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