Mechanistic insights into nitrogen fixation by nitrogenase enzymes

2015 ◽  
Vol 17 (44) ◽  
pp. 29541-29547 ◽  
Author(s):  
J. B. Varley ◽  
Y. Wang ◽  
K. Chan ◽  
F. Studt ◽  
J. K. Nørskov
Keyword(s):  
Nitrogen Fixation ◽  
High Temperatures ◽  
Sulfur Atom ◽  
Biological Nitrogen Fixation ◽  
Ammonia Synthesis ◽  
Catalytic Site ◽  
Bosch Process ◽  
Edge Site ◽  
Biological Nitrogen

The active catalytic site for biological nitrogen fixation is identified as an Fe-edge site underneath a vacated belt-sulfur atom (μ2 S) of the FeMoco cluster in nitrogenase. The evolution of the μ2 S as H2S is critical to electrochemically activating the inert N2, while its readsorption is required to dissociate the strongly bound NH3*. The reversible hinge-like behavior of the μ2 S provides an analog to the high temperatures and pressures required in industrial ammonia synthesis in the Haber–Bosch process.

scholarly journals Functional analysis of multiple nifB genes of Paenibacillus strains in synthesis of Mo-, Fe- and V-nitrogenases

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Qin Li ◽  
Haowei Zhang ◽  
Liqun Zhang ◽  
Sanfeng Chen
Keyword(s):  
Functional Analysis ◽  
Nitrogen Fixation ◽  
Gene Cluster ◽  
Biological Nitrogen Fixation ◽  
Deletion Analysis ◽  
Phylogeny Analysis ◽  
Biological Nitrogen

Abstract Background Biological nitrogen fixation is catalyzed by Mo-, V- and Fe-nitrogenases that are encoded by nif, vnf and anf genes, respectively. NifB is the key protein in synthesis of the cofactors of all nitrogenases. Most diazotrophic Paenibacillus strains have only one nifB gene located in a compact nif gene cluster (nifBHDKENX(orf1)hesAnifV). But some Paenibacillus strains have multiple nifB genes and their functions are not known. Results A total of 138 nifB genes are found in the 116 diazotrophic Paenibacillus strains. Phylogeny analysis shows that these nifB genes fall into 4 classes: nifBI class including the genes (named as nifB1 genes) that are the first gene within the compact nif gene cluster, nifBII class including the genes (named as nifB2 genes) that are adjacent to anf or vnf genes, nifBIII class whose members are designated as nifB3 genes and nifBIV class whose members are named as nifB4 genes are scattered on genomes. Functional analysis by complementation of the ∆nifB mutant of P. polymyxa which has only one nifB gene has shown that both nifB1 and nifB2 are active in synthesis of Mo-nitrogenase, while nifB3 and nifB4 genes are not. Deletion analysis also has revealed that nifB1 of Paenibacillus sabinae T27 is involved in synthesis of Mo-nitrogenase, while nifB3 and nifB4 genes are not. Complementation of the P. polymyxa ∆nifBHDK mutant with the four reconstituted operons: nifB1anfHDGK, nifB2anfHDGK, nifB1vnfHDGK and nifB2vnfHDGK, has shown both that nifB1 and nifB2 were able to support synthesis of Fe- or V-nitrogenases. Transcriptional results obtained in the original Paenibacillus strains are consistent with the complementation results. Conclusions The multiple nifB genes of the diazotrophic Paenibacillus strains are divided into 4 classes. The nifB1 located in a compact nif gene cluster (nifBHDKENX(orf1)hesAnifV) and the nifB2 genes being adjacent to nif or anf or vnf genes are active in synthesis of Mo-, Fe and V-nitrogenases, but nifB3 and nifB4 are not. The reconstituted anf system comprising 8 genes (nifBanfHDGK and nifXhesAnifV) and vnf system comprising 10 genes (nifBvnfHDGKEN and nifXhesAnifV) support synthesis of Fe-nitrogenase and V-nitrogenase in Paenibacillus background, respectively.

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