Nitrogenase MoFe protein subunits from Klebsiella pneumoniae expressed in foreign hosts. Characteristics and interactions.

Holland, D.B.; Zilberstein, Aviah; Govezensky, D; Salomon, Daniela; Zamir, Ada

doi:10.1016/s0021-9258(18)47487-x

Cited by 18 publications

(3 citation statements)

References 18 publications

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“…Pioneering efforts to express nif genes in the eukaryotic organisms Saccharomyces cerevisiae [16][17][18][19] and Chlamydomonas reinhardtii [20] are worth mentioning. Unfortunately, expression of nifH, or co-expression of nifD and nifK, in S. cerevisiae yielded inactive proteins lacking their metal clusters.…”

Section: Identified Barriersmentioning

confidence: 99%

Challenges to develop nitrogen-fixing cereals by direct nif-gene transfer

2014

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Section: Identified Barriersmentioning

confidence: 99%

Challenges to develop nitrogen-fixing cereals by direct nif-gene transfer

2014

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“…As early as 1972, researchers successfully transferred nif gene clusters into Escherichia coli and expressed active nitrogenase . Subsequently, research has continued to achieve heterologous expression of nif gene clusters in microorganisms such as yeast and cyanobacteria. − However, the expression activity of nitrogenase in these heterologous hosts often falls short of expectations and cannot be used in practical applications like crop production.…”

Section: Introductionmentioning

confidence: 99%

Deciphering the Nitrogen Fixation Gene Cluster in Vibrio natriegens: A Study on Optimized Expression and Application of Nitrogenase

He,

Zheng,

Xiao

et al. 2024

J. Agric. Food Chem.

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Microbial nitrogen fixation presents a viable alternative to chemical fertilizers, yet the limited colonization and specificity of naturally occurring nitrogen-fixing microorganisms present significant challenges to their widespread application. In this study, we identified a nitrogen fixation gene cluster (VNnif) in Vibrio natriegens (VN) and tested its nitrogenase activity through the acetylene reduction assay. We investigated the potential utilization of nitrogenase by incorporating the nitrogenase gene cluster from VN into plant growth-promoting rhizosphere bacteria Pseudomonas protegens CHA0 and enhancing its activity to 48.16 nmol C2H2/mg/h through promoter replacement and cluster rearrangement. The engineered strain CHA0-PVNnif was found to positively impact the growth of Arabidopsis thaliana col-0 and Triticum aestivum L. (wheat). This study expanded the role of plant growth-promoting rhizobacteria (PGPR) and provided a research foundation for enhancing nitrogenase activity.

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“…S. cerevisiae is also a key model organism to test eukaryotic expression of Nif proteins, as its cells can be grown microaerobically. Pioneering efforts to express nif genes of Klebsiella oxytoca in S. cerevisiae were performed. − Recently, the active NifH was produced in S. cerevisiae cells by expressing the 4 genes ( nifH , nifU , nifS , nifM ) of Azotobacter vinelandii . The active NifB (cofactor maturase) of nitrogenase was formed in yeast by expressing Methanocaldococcus infernus nifB together with nifU , nifS , and fdxN genes that are involved in NifB [Fe–S] cluster assembly and activity .…”

mentioning

confidence: 99%

Combined Assembly and Targeted Integration of Multigene for Nitrogenase Biosynthetic Pathway in Saccharomyces cerevisiae

et al. 2019

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Biological nitrogen fixation, a process unique to diazotrophic prokaryote, is catalyzed by the nitrogenase complex. There has been a long-standing interest in reconstituting a nitrogenase biosynthetic pathway in a eukaryotic host with the final aim of developing N2-fixing cereal crops. In this study, we report that a nitrogenase biosynthetic pathway (∼38 kb containing 15 genes) was assembled in two individual one-step methods via in vivo assembly and integrated at δ and HO sites in Saccharomyces cerevisiae chromosome. Of the 15 genes, 11 genes (nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA, nifV, groES, groEL) were from Paenibacillus polymyxa WLY78 and 4 genes (nifS, nifU, nifF, nifJ) were from Klebsiella oxytoca. The 15-gene nitrogenase biosynthetic pathway was correctly assembled and transcribed in the recombinant S. cerevisiae. The NifDK tetramer with an identical molecular weight as that of P. polymyxa was formed in yeast and the expressed NifH exhibited the activity of Fe protein. This study demonstrates that it will be possible to produce active nitrogenase in eukaryotic hosts.

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Nitrogenase MoFe protein subunits from Klebsiella pneumoniae expressed in foreign hosts. Characteristics and interactions.

Cited by 18 publications

References 18 publications

Challenges to develop nitrogen-fixing cereals by direct nif-gene transfer

Challenges to develop nitrogen-fixing cereals by direct nif-gene transfer

Deciphering the Nitrogen Fixation Gene Cluster in Vibrio natriegens: A Study on Optimized Expression and Application of Nitrogenase

Combined Assembly and Targeted Integration of Multigene for Nitrogenase Biosynthetic Pathway in Saccharomyces cerevisiae

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