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Published VersionCC BY V4.0, Open Access
Journal article
The structural components of the Azotobacter vinelandii iron-only nitrogenase, AnfDKG, form a protein complex within the plant mitochondrial matrix
Plant Molecular Biology, Vol.112, pp.279-291
2023
PMID: 37326800
Abstract
A long-held goal of synthetic biology has been the transfer of a bacterial nitrogen-fixation pathway into plants to reduce the use of chemical fertiliser on crops such as rice, wheat and maize. There are three classes of bacterial nitrogenase, named after their metal requirements, containing either a MoFe-, VFe- or FeFe-cofactor, that converts N-2 gas to ammonia. Relative to the Mo-nitrogenase the Fe-nitrogenase is not as efficient for catalysis but has less complex genetic and metallocluster requirements, features that may be preferable for engineering into crops. Here we report the successful targeting of bacterial Fe-nitrogenase proteins, AnfD, AnfK, AnfG and AnfH, to plant mitochondria. When expressed as a single protein AnfD was mostly insoluble in plant mitochondria, but coexpression of AnfD with AnfK improved its solubility. Using affinity-based purification of mitochondrially expressed AnfK or AnfG we were able to demonstrate a strong interaction of AnfD with AnfK and a weaker interaction of AnfG with AnfDK. This work establishes that the structural components of the Fe-nitrogenase can be engineered into plant mitochondria and form a complex, which will be a requirement for function. This report outlines the first use of Fe-nitrogenase proteins within a plant as a preliminary step towards engineering an alternative nitrogenase into crops.
Details
- Title
- The structural components of the Azotobacter vinelandii iron-only nitrogenase, AnfDKG, form a protein complex within the plant mitochondrial matrix
- Authors
- E Johnston (Author) - CSIRO Environment (Australia)S Okada (Author) - CSIRO Environment (Australia)C M Gregg (Author) - CSIRO Agriculture and Food (Australia)A C Warden (Author) - CSIRO Environment (Australia)V Rolland (Author) - CSIRO Agriculture and Food (Australia)V Gillespie (Author) - CSIRO Agriculture and Food (Australia)K Byrne (Author) - CSIRO Agriculture and Food (Australia)Michelle Colgrave (Author) - CSIRO Agriculture and Food (Australia)Andrew Eamens (Author) - University of the Sunshine Coast, Queensland, School of Health - BiomedicineRobert Silas Allen (Corresponding Author) - CSIRO Agriculture and Food (Australia)C C Wood (Author) - CSIRO Agriculture and Food (Australia)
- Publication details
- Plant Molecular Biology, Vol.112, pp.279-291
- Publisher
- Springer Dordrecht
- Date published
- 2023
- DOI
- 10.1007/s11103-023-01363-3
- ISSN
- 1573-5028; 0167-4412
- PMID
- 37326800
- Copyright note
- This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
- Organisation Unit
- School of Health - Biomedicine
- Language
- English
- Record Identifier
- 99968898102621
- Output Type
- Journal article
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- Biochemistry & Molecular Biology
- Plant Sciences
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