99130 20230519145448.0 doi 10.1038/s42003-020-01536-6 sideral 122581 ART-2021-122581 eng Jiang, X. Exploiting genetic diversity and gene synthesis to identify superior nitrogenase NifH protein variants to engineer N2-fixation in plants 2021 Access copy available to the general public Unrestricted Engineering nitrogen fixation in eukaryotes requires high expression of functional nitrogenase structural proteins, a goal that has not yet been achieved. Here we build a knowledge-based library containing 32 nitrogenase nifH sequences from prokaryotes of diverse ecological niches and metabolic features and combine with rapid screening in tobacco to identify superior NifH variants for plant mitochondria expression. Three NifH variants outperform in tobacco mitochondria and are further tested in yeast. Hydrogenobacter thermophilus (Aquificae) NifH is isolated in large quantities from yeast mitochondria and fulfills NifH protein requirements for efficient N2 fixation, including electron transfer for substrate reduction, P-cluster maturation, and FeMo-co biosynthesis. H. thermophilus NifH expressed in tobacco leaves shows lower nitrogenase activity than that from yeast. However, transfer of [Fe4S4] clusters from NifU to NifH in vitro increases 10-fold the activity of the tobacco-isolated NifH, revealing that plant mitochondria [Fe-S] cluster availability constitutes a bottleneck to engineer plant nitrogenases. info:eu-repo/grantAgreement/ES/DGA/E35-17R info:eu-repo/grantAgreement/ES/DGA-FEDER/Construyendo Europa desde Aragón info:eu-repo/grantAgreement/ES/MINECO/CTQ2015-64486-R info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 6.548 2021 BIOLOGY 15 / 94 = 0.16 2021 Q1 T1 2.256 2021 Biochemistry, Genetics and Molecular Biology (miscellaneous) 2021 Q1 Agricultural and Biological Sciences (miscellaneous) 2021 Q1 6.0 2021 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Payá-Tormo, L. Coroian, D. García-Rubio, I. Universidad de Zaragoza (orcid)0000-0002-1827-1250 Castellanos-Rueda, R. Eseverri, Á. López-Torrejón, G. Burén, S. Rubio, L.M. 2003 395 Universidad de Zaragoza Dpto. Física Materia Condensa. Área Física Materia Condensada 4, 1 (2021), 4 [11 pp] Communications Biology 2399-3642 562032 http://zaguan.unizar.es/record/99130/files/texto_completo.pdf Versión publicada 1560932 http://zaguan.unizar.es/record/99130/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:99130 articulos driver 2023-05-18-14:40:02 ARTICLE