Nitrogenase Cofactor Maturase NifB Isolated from Transgenic Rice is Active in FeMo-co Synthesis

The engineering of nitrogen fixation in plants requires assembly of an active prokaryotic nitrogenase complex, which is yet to be achieved. Nitrogenase biogenesis relies on NifB, which catalyzes the formation of the [8Fe−9S−C] metal cluster NifB-co. This is the first committed step in the biosynthes...

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Autores: He, Wenshu, Burén, Stefan, Baysal, Can, Jiang, Xi, Capell Capell, Teresa, Christou, Paul, Rubio, Luis
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Universitat de Lleida (UdL)
Repositorio:Repositori Obert UdL
OAI Identifier:oai:repositori.udl.cat:10459.1/84460
Acceso en línea:https://doi.org/10.1021/acssynbio.2c00194
http://hdl.handle.net/10459.1/84460
Access Level:acceso abierto
Palabra clave:Nitrogen fixation
Transgenic rice
NifB-co
Synthetic biology
Iron-molybdenum cofactor
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spelling Nitrogenase Cofactor Maturase NifB Isolated from Transgenic Rice is Active in FeMo-co SynthesisHe, WenshuBurén, StefanBaysal, CanJiang, XiCapell Capell, TeresaChristou, PaulRubio, LuisNitrogen fixationTransgenic riceNifB-coSynthetic biologyIron-molybdenum cofactorThe engineering of nitrogen fixation in plants requires assembly of an active prokaryotic nitrogenase complex, which is yet to be achieved. Nitrogenase biogenesis relies on NifB, which catalyzes the formation of the [8Fe−9S−C] metal cluster NifB-co. This is the first committed step in the biosynthesis of the iron−molybdenum cofactor (FeMo-co) found at the nitrogenase active site. The production of NifB in plants is challenging because this protein is often insoluble in eukaryotic cells, and its [Fe−S] clusters are extremely unstable and sensitive to O2. As a first step to address this challenge, we generated transgenic rice plants expressing NifB from the Archaea Methanocaldococcus infernus and Methanothermobacter thermautotrophicus. The recombinant proteins were targeted to the mitochondria to limit exposure to O2 and to have access to essential [4Fe−4S] clusters required for NifB-co biosynthesis. M. infernus and M. thermautotrophicus NifB accumulated as soluble proteins in planta, and the purified proteins were functional in the in vitro FeMo-co synthesis assay. We thus report NifB protein expression and purification from an engineered staple crop, representing a first step in the biosynthesis of a functional NifDK complex, as required for independent biological nitrogen fixation in cereals.American Chemical Society2022info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttps://doi.org/10.1021/acssynbio.2c00194http://hdl.handle.net/10459.1/84460reponame:Repositori Obert UdL instname:Universitat de Lleida (UdL)InglésReproducció del document publicat a https://doi.org/10.1021/acssynbio.2c00194ACS Synthetic Biology, 2022, vol. 11, núm. 9, p. 3028-3036.cc-by (c) Wenshu He et al., 2022info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/4.0/oai:repositori.udl.cat:10459.1/844602026-06-24T12:42:17Z
dc.title.none.fl_str_mv Nitrogenase Cofactor Maturase NifB Isolated from Transgenic Rice is Active in FeMo-co Synthesis
title Nitrogenase Cofactor Maturase NifB Isolated from Transgenic Rice is Active in FeMo-co Synthesis
spellingShingle Nitrogenase Cofactor Maturase NifB Isolated from Transgenic Rice is Active in FeMo-co Synthesis
He, Wenshu
Nitrogen fixation
Transgenic rice
NifB-co
Synthetic biology
Iron-molybdenum cofactor
title_short Nitrogenase Cofactor Maturase NifB Isolated from Transgenic Rice is Active in FeMo-co Synthesis
title_full Nitrogenase Cofactor Maturase NifB Isolated from Transgenic Rice is Active in FeMo-co Synthesis
title_fullStr Nitrogenase Cofactor Maturase NifB Isolated from Transgenic Rice is Active in FeMo-co Synthesis
title_full_unstemmed Nitrogenase Cofactor Maturase NifB Isolated from Transgenic Rice is Active in FeMo-co Synthesis
title_sort Nitrogenase Cofactor Maturase NifB Isolated from Transgenic Rice is Active in FeMo-co Synthesis
dc.creator.none.fl_str_mv He, Wenshu
Burén, Stefan
Baysal, Can
Jiang, Xi
Capell Capell, Teresa
Christou, Paul
Rubio, Luis
author He, Wenshu
author_facet He, Wenshu
Burén, Stefan
Baysal, Can
Jiang, Xi
Capell Capell, Teresa
Christou, Paul
Rubio, Luis
author_role author
author2 Burén, Stefan
Baysal, Can
Jiang, Xi
Capell Capell, Teresa
Christou, Paul
Rubio, Luis
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Nitrogen fixation
Transgenic rice
NifB-co
Synthetic biology
Iron-molybdenum cofactor
topic Nitrogen fixation
Transgenic rice
NifB-co
Synthetic biology
Iron-molybdenum cofactor
description The engineering of nitrogen fixation in plants requires assembly of an active prokaryotic nitrogenase complex, which is yet to be achieved. Nitrogenase biogenesis relies on NifB, which catalyzes the formation of the [8Fe−9S−C] metal cluster NifB-co. This is the first committed step in the biosynthesis of the iron−molybdenum cofactor (FeMo-co) found at the nitrogenase active site. The production of NifB in plants is challenging because this protein is often insoluble in eukaryotic cells, and its [Fe−S] clusters are extremely unstable and sensitive to O2. As a first step to address this challenge, we generated transgenic rice plants expressing NifB from the Archaea Methanocaldococcus infernus and Methanothermobacter thermautotrophicus. The recombinant proteins were targeted to the mitochondria to limit exposure to O2 and to have access to essential [4Fe−4S] clusters required for NifB-co biosynthesis. M. infernus and M. thermautotrophicus NifB accumulated as soluble proteins in planta, and the purified proteins were functional in the in vitro FeMo-co synthesis assay. We thus report NifB protein expression and purification from an engineered staple crop, representing a first step in the biosynthesis of a functional NifDK complex, as required for independent biological nitrogen fixation in cereals.
publishDate 2022
dc.date.none.fl_str_mv 2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://doi.org/10.1021/acssynbio.2c00194
http://hdl.handle.net/10459.1/84460
url https://doi.org/10.1021/acssynbio.2c00194
http://hdl.handle.net/10459.1/84460
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a https://doi.org/10.1021/acssynbio.2c00194
ACS Synthetic Biology, 2022, vol. 11, núm. 9, p. 3028-3036.
dc.rights.none.fl_str_mv cc-by (c) Wenshu He et al., 2022
info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by/4.0/
rights_invalid_str_mv cc-by (c) Wenshu He et al., 2022
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv reponame:Repositori Obert UdL
instname:Universitat de Lleida (UdL)
instname_str Universitat de Lleida (UdL)
reponame_str Repositori Obert UdL
collection Repositori Obert UdL
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.811543

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