Observatorio de I+D+i UPM

Memorias de investigación
Communications at congresses:
Expression of the nitrogenase iron-molybdenum cofactor maturase NifB in mitocondria of Saccharomyces cerevisiae.
Year:2017
Research Areas
  • Microbiology,
  • Biotechnology
Information
Abstract
Active NifB is a milestone in the process of engineering nitrogen fixing plants. NifB is an extremely O2-sensitive S-adenosyl methionine (SAM)?radical enzyme that provides the key metal cluster intermediate (NifB-co) for the biosyntheses of the active-site cofactors of all three types of nitrogenases. NifB and NifB-co are unique to diazotrophic organisms. In this work, we have expressed synthetic codon-optimized versions of NifB from the ?-proteobacterium Azotobacter vinelandii and the thermophilic methanogen Methanocaldococcus infernus in Saccharomyces cerevisiae and in Nicotiana benthamiana. NifB proteins were targeted to the mitochondria, where O2 consumption is high and bacterial-like [Fe-S] cluster assembly operates. In yeast, NifB proteins were co-expressed with NifU, NifS, and FdxN proteins that are involved in NifB [Fe?S] cluster assembly and activity. The synthetic version of thermophilic NifB accumulated in soluble form within the yeast cell, while the A. vinelandii version appeared to form aggregates. Similarly, NifB from M. infernus was expressed at higher levels in leaves of Nicotiana benthamiana and accumulated as a soluble protein while A. vinelandii NifB was mainly associated with the non-soluble cell fraction. Soluble M. infernus NifB was purified from aerobically grown yeast and biochemically characterized. The purified protein was functional in the in vitro FeMo-co synthesis assay. This work presents the first active NifB protein purified from a eukaryotic cell, and highlights the importance of screening nif genes from different organisms in order to sort the best candidates to assemble a functional plant nitrogenase.
International
Si
Congress
20th International Congress on Nitrogen Fixation. Internacional
970
Place
Granada
Reviewers
Si
ISBN/ISSN
00000000
Start Date
03/09/2017
End Date
07/09/2017
From page
25
To page
25
Expression of the nitrogenase iron-molybdenum cofactor maturase NifB in mitocondria of Saccharomyces cerevisiae.
Participants
  • Autor: Nils Stefan Buren (UPM)
  • Autor: Gema Lopez Torrejon (UPM)
  • Autor: Carlos Echavarri Erasun (UPM)
  • Autor: Marcel Veldhuizen . (UPM)
  • Autor: José María Buesa Galiano (UPM)
  • Autor: Luis Manuel Rubio Herrero (UPM)
Research Group, Departaments and Institutes related
  • Creador: Grupo de Investigación: BIOLOGÍA MOLECULAR Y COMPUTACIONAL
  • Centro o Instituto I+D+i: Centro de Biotecnología y Genómica de Plantas, CBGP
  • Departamento: Biotecnología - Biología Vegetal
S2i 2020 Observatorio de investigación @ UPM con la colaboración del Consejo Social UPM
Cofinanciación del MINECO en el marco del Programa INNCIDE 2011 (OTR-2011-0236)
Cofinanciación del MINECO en el marco del Programa INNPACTO (IPT-020000-2010-22)