PreprintCited by 5Genomics and Phylogenetic StudiesMicrobial Community Ecology and PhysiologyEnzyme Structure and Function

EcoFoldDB: Protein structure-guided functional profiling of ecologically relevant microbial traits at the metagenome scale

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Ghaly, Timothy M., Rajabal, Vaheesan, Russell, Dylan, Colombi, Elena, Tetu, Sasha G.·2025·Environmental Microbiology

Preprint — 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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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)
OpenAlex
W4414301061
Cited by
5
References
120
Citations by year
2026: 3 · 2025: 2
FWCI
1.50
Type
journal article
Language
en
Open access
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

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