Observatorio de I+D+i UPM

Memorias de investigación
Communications at congresses:
Diversity and functional analyses of NifB-cofactor maturase (NifB) for its implementation in eukaryotic systems
Year:2017
Research Areas
  • Microbiology,
  • Biotechnology
Information
Abstract
One of the main hurdles to engineer nitrogenase in a non-diazotrophic host is achieving NifB activity. NifB is an extremely unstable and oxygen sensitive protein that catalyzes a low-potential SAM-radical dependent reaction. The product of NifB activity is called NifB-co, a complex [8Fe-9S-C] cluster that serves as obligate intermediate in the biosyntheses of the active-site cofactors of all known nitrogenases. Here we study the diversity and phylogeny of naturally occurring NifB proteins, their protein architecture and the functions of the distinct NifB domains in order to understand what defines a catalytically active NifB. Focus is on NifB from the thermophile Chlorobium tepidum (two-domain architecture), the hyperthermophile Methanocaldococcus infernus (single-domain architecture) and the mesophile Klebsiella oxytoca (two-domain architecture), showing in silico characterization of their nitrogen fixation (nif) gene clusters, conserved NifB motifs, and functionality. C. tepidum and M. infernus NifB were able to complement an Azotobacter vinelandii (?nifB) mutant restoring the Nif+ phenotype and thus demonstrating their functionality in vivo. In addition, purified C. tepidum NifB exhibited activity in the in vitro NifB-dependent nitrogenase reconstitution assay. Intriguingly, changing the two-domain K. oxytoca NifB to single-domain by removal of the C-terminal NifX-like extension resulted in higher in vivo nitrogenase activity, demonstrating that this domain is not required for nitrogen fixation in mesophiles.
International
Si
Congress
20th International Congress on Nitrogen Fixation
970
Place
Granada
Reviewers
Si
ISBN/ISSN
00000000
Start Date
03/09/2017
End Date
09/09/2017
From page
25
To page
25
Diversity and functional analyses of NifB-cofactor maturase (NifB) for its implementation in eukaryotic systems
Participants
  • Autor: Carlos Echavarri Erasun (UPM)
  • Autor: Emilio Jimenez Vicente (UPM)
  • Autor: Nils Stefan Buren (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
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