Diversity and Functional Analysis of the FeMo-Cofactor Maturase NifB

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dc.contributor.authorArragain, Simonen
dc.contributor.authorJimenez-Vicente, Emilioen
dc.contributor.authorScandurra, Alessandro A.en
dc.contributor.authorBuren, Stefanen
dc.contributor.authorRubio, Luis M.en
dc.contributor.authorEchavarri-Erasun, Carlosen
dc.contributor.departmentBiochemistryen
dc.date.accessioned2019-04-24T18:41:39Zen
dc.date.available2019-04-24T18:41:39Zen
dc.date.issued2017-11-14en
dc.description.abstractOne 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 (singledomain 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 (Delta 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.en
dc.description.notesFunding for this research was provided by Bill & Melinda Gates Foundation OPP1143172, ERC Starting Grant 205442, and MINECO BIO2014-59131-R. AS was recipient of FPI Fellowship BES-2010-038322.en
dc.description.sponsorshipBill & Melinda Gates Foundation [OPP1143172]en
dc.description.sponsorshipERC Starting Grant [205442]en
dc.description.sponsorshipMINECO [BIO2014-59131-R]en
dc.description.sponsorshipFPI Fellowship [BES-2010-038322]en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.3389/fpls.2017.01947en
dc.identifier.eissn1664-462Xen
dc.identifier.other1947en
dc.identifier.pmid29250084en
dc.identifier.urihttp://hdl.handle.net/10919/89108en
dc.identifier.volume8en
dc.language.isoenen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectnitrogenaseen
dc.subjectiron-molybdenum cofactoren
dc.subjectSAM-radicalen
dc.subjectnitrogen fixationen
dc.subjectAzotobacteren
dc.subjectmethanogensen
dc.titleDiversity and Functional Analysis of the FeMo-Cofactor Maturase NifBen
dc.title.serialFrontiers In Plant Scienceen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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