Evidence map›Paper›PMID 40831097›Full record

ArticlePlant biotechnology journal2025

Computationally Guided Design of a Soluble, Abundant and Functional AnfH Variant for the Expression of Fe-Only Nitrogenase in Plant Mitochondria.

Christina M Gregg, Barnabas Gall, Xueqin Wang, Shoko Okada, Trevor D Rapson, Robert S Allen, Ingrid Venables, Anu Mathew, Craig C Wood

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Plant biotechnology journal, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

9 authors.

Christina M GreggAgriculture and Food, Commonwealth Scientific and Industrial Research Organisation, Canberra, Australian Capital Territory, Australia.ORCID https://orcid.org/0000-0003-4440-4624
Barnabas GallResearch School of Chemistry, Australian National University, Canberra, Australian Capital Territory, Australia.ORCID https://orcid.org/0009-0003-3182-9911
Xueqin WangAgriculture and Food, Commonwealth Scientific and Industrial Research Organisation, Canberra, Australian Capital Territory, Australia.ORCID https://orcid.org/0009-0000-0635-1870
Shoko OkadaEnvironment, Commonwealth Scientific and Industrial Research Organisation, Canberra, Australian Capital Territory, Australia.ORCID https://orcid.org/0000-0002-1759-2677
Trevor D RapsonAgriculture and Food, Commonwealth Scientific and Industrial Research Organisation, Canberra, Australian Capital Territory, Australia.ORCID https://orcid.org/0000-0003-1289-0146
Robert S AllenAgriculture and Food, Commonwealth Scientific and Industrial Research Organisation, Canberra, Australian Capital Territory, Australia.ORCID https://orcid.org/0000-0002-2994-2996
Ingrid VenablesAgriculture and Food, Commonwealth Scientific and Industrial Research Organisation, Canberra, Australian Capital Territory, Australia.ORCID https://orcid.org/0000-0002-5258-6563
Anu MathewAgriculture and Food, Commonwealth Scientific and Industrial Research Organisation, Canberra, Australian Capital Territory, Australia.ORCID https://orcid.org/0009-0009-5384-1288
Craig C WoodAgriculture and Food, Commonwealth Scientific and Industrial Research Organisation, Canberra, Australian Capital Territory, Australia.ORCID https://orcid.org/0000-0002-6425-0434

Funding

Commonwealth Scientific and Industrial Research OrganisationCotton Seed Distributors Limited
6 · The paper itself

Abstract

Engineering nitrogenase directly into crops is a long-held aspiration in plant biotechnology. Of the three types, the Fe-only nitrogenase is a promising engineering target, as it has a simpler maturation pathway than the MoFe- and VFe-nitrogenases and does not require any heterometals for its cofactor. However, previously, we have reported that the obligate electron donor of the Fe-only nitrogenase from A. vinelandii, AnfH, is mostly insoluble when expressed in plant mitochondria. Here, we employed computational methods, the Protein Repair One-Stop Shop (PROSS) protein optimisation algorithm and Rosetta energy calculations, to design eight variants of AnfH with improved soluble expression. The amino acid substitutions chosen were predicted to lower the free energy of the native state of the protein. All eight AnfH variants, containing between 1 and 11 amino acid substitutions, were more soluble than wild-type AnfH in plant mitochondria. Of these, three variants were isolated from N. benthamiana leaf, of which AnfH variant 6 (AnfH V6, T200A T228V E241H) had the best features, being approx. 90-fold more abundant in the soluble fraction. Importantly, AnfH V6 was also functional in

Indexed as

Azotobacter vinelandiiMitochondriaNitrogenaseNicotianaProtein EngineeringNitrogenaseAnfHcomputational designheterologous protein productioniron–sulfurmaturationmitochondrianitrogenasenitrogen fixationplantsolubility

Identifiers

PMID40831097
PMCPMC12665077

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.