EcoFoldDB: Protein structure-guided functional profiling of ecologically relevant microbial traits at the metagenome scale
Open accessPreprint — published as 10.1111/1462-2920.70178
Abstract
Microbial communities are fundamental to planetary health and ecosystem processes. High-throughput metagenomic sequencing has provided unprecedented insights into the structure and function of these communities. However, functionally profiling metagenomes remains constrained due to the limited sensitivity of existing sequence homology-based methods to annotate evolutionarily divergent genes. Protein structure, more conserved than sequence, and intrinsically tied to molecular function, offers a solution. Capitalising on recent breakthroughs in structural bioinformatics, we present EcoFoldDB, a database of protein structures curated for ecologically relevant microbial traits, and its companion pipeline, EcoFoldDB-annotate , which leverages Foldseek with the ProstT5 protein language model for rapid structural homology searching directly from sequence data. EcoFoldDB-annotate outperforms state-of-the-art sequence-based methods in annotating metagenomic proteins, in terms of sensitivity and precision. To demonstrate its utility and scalability, we performed structure-guided functional profiling of 32 million proteins encoded by 8,000 high-quality metagenome-assembled genomes from the global soil microbiome. EcoFoldDB-annotate could resolve the phylogenetic partitioning of important nitrogen cycling pathways, from taxonomically restricted nitrifiers to more widespread denitrifiers, as well as identifying novel, uncultivated bacterial taxa enriched in plant growth-promoting traits. We anticipate that EcoFoldDB will enable researchers to extract ecological insights from environmental genomes and metagenomes, and accelerate discoveries in microbial ecology.
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Unifying the known and unknown microbial coding sequence space
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Microbiology of the Anthropocene
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Role of d-Cysteine Desulfhydrase in the Adaptation of Escherichia coli to d-Cysteine
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A nitrite-oxidising bacterium constitutively consumes atmospheric hydrogen
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An ecological perspective on microbial genes of unknown function in soilPreprint
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Agriculture alters protein evolution of nitrogen cycling genes in soil bacteria at a global scalePreprint
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Self-citation
Flagged first, then by citations
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Adaptative and ancient co-evolution of integrons with Xanthomonas genomes
Colombi, E., Ghaly, T. M., Rajabal, V., Elbourne, L. D. H., Gillings, M. R., & Tetu, S. G. (2025) · Microbial Genomics · doi:10.1099/mgen.0.001503 · Cited by 1 · see citing works
Self-citationOumer, O. J., Simachew, A., Hartmann, E. M., Alexandersson, E., & Johansson, E. (2025) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2025.06.14.659726
Diverse conjugative and mobilisable elements underpin key adaptive traits in XanthomonasPreprint
Colombi, E., Ghaly, T. M., Samarakoon, N., Rajabal, V., & Tetu, S. G. (2026) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.64898/2026.06.24.734381
Self-citationTechniques in Molecular Diagnosis
Haq, I. U., Khan, N. A., Ijaz, S., & Ali, H. (2026) · Springer eBooks · doi:10.1007/978-981-92-2642-9_8
Soil microbial traits shift on contrasting timescales following restoration of former grazing lands
Ghaly, T. M., McPherson, V. J., Rajabal, V., Ghaly, M., Taws, N., Gallagher, R. V., Roux, J. J. L., & Tetu, S. G. (2026) · Journal of Environmental Management · doi:10.1016/j.jenvman.2026.131161
Self-citation
- Authors
- Ghaly, T. M., Rajabal, V., Russell, D., Colombi, E., & Tetu, S. G.
- Affiliations
- ARC Centre of Excellence in Synthetic Biology (AU)
- Corresponding author
- Ghaly, Timothy M.
- Venue
- Environmental Microbiology
- Publisher
- openRxiv
- Published
- 2025-04-02
- Volume
- 27
- Issue
- 9
- Pages
- e70178-e70178
- ISSN
- 1462-2912 (ISSN-L)
- PMC
- PMC12441223
- OpenAlex
- W4414301061
- Cited by
- 5
- References
- 120
- Citations by year
- 2026: 3 · 2025: 2
- FWCI
- 1.50
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Environmental Microbiology (publishedVersion, cc-by-nc-nd)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nd)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- Funders
- Macquarie University
- Australian Research Council (CE200100029)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
- Physical Sciences › Environmental Science › Ecology › Microbial Community Ecology and Physiology
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Keywords
- protein structure database, Foldseek, soil microbiome, metagenome-assembled genomes, nitrogen cycling, nitrifying bacteria, denitrifiers, plant growth-promoting bacteria, protein language models
- MeSH terms
- Bacteria, Bacterial Proteins, Phylogeny, Protein Conformation, Soil Microbiology, Computational Biology, Databases, Protein, Metagenome, Metagenomics, Microbiota
About these checks
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