Abstract Summary Gene annotation of metagenome-assembled genomes is a critical step in determining the functional potential of microbial communities from environmental samples. However, annotation workflows using tools such as Prokka or Bakta produce per-bin output with 10 to 14 files per bin, making manual review infeasible at scale. Existing tools incompletely aggregate and visualize gene annotation content across an entire metagenomic dataset. Here we present annoreport, a single-script Python tool requiring no external dependencies beyond Python 3.9+ that accepts output from either Prokka or Bakta, automatically detecting the annotation tool used. annoreport produces an interactive web-based report summarizing gene product frequencies, hypothetical protein rates, feature type distributions, and functional gene clustering via UniProt annotation across all bins. Applied to 206 metagenome-assembled genomes from Antarctic soil metagenomes, annoreport identified 603,799 coding sequences with a 47.1% annotation rate and revealed functional categorization in Transport & Membrane, Nucleotide Binding, and DNA Metabolism categories. Availability and implementation Freely available at https://github.com/keplerridge/annoreport under MIT license, via Bioconda (annoreport) and PyPI (annoreport).
This study presents the first large-scale investigation of four prominent open-source annotation tools (Prokka, Bakta, EggNOG-mapper, and PGAP) across 156,033 diverse genomes, highlighting tool-specific strengths crucial for selecting optimal solutions based on genome quality, taxonomy, and origin.
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FEDKEA, an enzyme annotation tool leveraging protein language models, and a user-friendly, FEDKEA-based metagenomic pipeline, MEnzMap, which encompasses the entire analysis workflow—from raw data quality control to function prediction and downstream analyses are designed.
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Prokanota provides an accessible and reproducible annotation backbone while allowing project-specific databases and evidence layers to be incorporated without modifying workflow code, and fills a practical gap between fixed, standardized annotation tools and fully custom expert-built workflows.
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The results establish Sma3s v3 as a scalable and interpretable tool for functional annotation and re-annotation of proteomes, pangenomes, and metagenomic protein catalogues.
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The GenomeCompendium is released, a public database and interactive analysis tool for complete prokaryotic genomes and it is shown that complex, repeat-rich genomes are more common than previously estimated.
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This review presents a practical, workflow-oriented guide to microbiome data analysis, from raw DNA sequence processing to statistical interpretation and biological insight, and highlights emerging technologies, including machine learning methods that are beginning to reshape the field.
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