Biosynthesis of cofactor‐activatable iron‐only nitrogenase in Saccharomyces cerevisiae

Summary Engineering nitrogenase in eukaryotes is hampered by its genetic complexity and by the oxygen sensitivity of its protein components. Of the three types of nitrogenases, the Fe‐only nitrogenase is considered the simplest one because its function depends on fewer gene products than the homolog...

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Bibliographic Details
Published in:Microbial biotechnology 2021-05, Vol.14 (3), p.1073-1083
Main Authors: López‐Torrejón, Gema, Burén, Stefan, Veldhuizen, Marcel, Rubio, Luis M.
Format: Article
Language:English
Subjects:
Acetylene
Acetylene reduction
Antibodies
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Biosynthesis
Chloroplasts
Complexity
E coli
Eukaryotes
Gene expression
Homology
Iron
Medical research
Mitochondria
Molybdenum
Nitrogen Fixation
Nitrogenase
Nitrogenase - metabolism
Nitrogenation
Oxygen
Proteins
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Yeast
Yeasts
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Summary:Summary Engineering nitrogenase in eukaryotes is hampered by its genetic complexity and by the oxygen sensitivity of its protein components. Of the three types of nitrogenases, the Fe‐only nitrogenase is considered the simplest one because its function depends on fewer gene products than the homologous and more complex Mo and V nitrogenases. Here, we show the expression of stable Fe‐only nitrogenase component proteins in the low‐oxygen mitochondria matrix of S. cerevisiae. As‐isolated Fe protein (AnfH) was active in electron donation to NifDK to reduce acetylene into ethylene. Ancillary proteins NifU, NifS and NifM were not required for Fe protein function. The FeFe protein existed as apo‐AnfDK complex with the AnfG subunit either loosely bound or completely unable to interact with it. Apo‐AnfDK could be activated for acetylene reduction by the simple addition of FeMo‐co in vitro, indicating preexistence of the P‐clusters even in the absence of coexpressed NifU and NifS. This work reinforces the use of Fe‐only nitrogenase as simple model to engineer nitrogen fixation in yeast and plant mitochondria. Fe‐only nitrogenase components were expressed and matured in S. cerevisiae mitochondrial. The electron donor component was active as isolated and the catalytic component was ready for activation by cofactor insertion.
ISSN:1751-7915
1751-7915
DOI:10.1111/1751-7915.13758