title: Expression and purification of NifB proteins from aerobic and anaerobic sources.
abstract: NifB is the key protein in the biosynthesis of nitrogenase iron-molybdenum cofactor. Due to its extreme sensitivity to O2 and inherent protein instability, NifB proteins must be purified under strict anaerobic conditions by using affinity chromatography methods. We describe here the methods for NifB purification from cells of the strict aerobic nitrogen-fixing bacterium Azotobacter vinelandii, the facultative anaerobic nitrogen-fixing bacterium Klebsiella pneumoniae, and the facultative anaerobic non-nitrogen fixing bacterium Escherichia coli recombinantly expressing a nifB gene of thermophilic origin.
authors.affiliation: Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid, Campus de Montegancedo, Pozuelo de Alarcón, 28223, Madrid, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Echavarri-Erasun
authors.forename: Simon
authors.initials: S
authors.lastname: Arragain
authors.forename: Alessandro A
authors.initials: AA
authors.lastname: Scandurra
authors.forename: Luis M
authors.initials: LM
authors.lastname: Rubio
mesh_terms: D000332:Aerobiosis; D000693:Anaerobiosis; D016948:Azotobacter vinelandii / Q000378:metabolism*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000302:isolation & purification*; D001671:Biochemistry / Q000379:methods*; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000378:metabolism*; D006639:Histidine; D007711:Klebsiella pneumoniae / Q000378:metabolism*; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; C471213:His-His-His-His-His-His; C092498:NifB protein, Bacteria; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D006639:Histidine