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Protein database construction from de novo peptide evidence for proteomics and metaproteomics.

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FastaLake

Read the paper — bioRxiv · DOI: 10.64898/2026.10.08.757395

Release CI Python Licence: MIT

Install · Quickstart · Documentation · Examples · Paper code

FastaLake uses de novo peptide predictions to select compact protein databases for metaproteomics. It retrieves complete proteins from a large sequence reservoir, selects a database for each mass spectrometry acquisition, and runs Sage to identify peptides. Study-wide protein grouping and quantification turn those results into a matrix ready for analysis.

Spectra and de novo predictions guide protein selection from a sequence reservoir, followed by database searching and study-level quantification.

Features

  • Select complete proteins from public sequence catalogues using exact peptide lookup.
  • Search and quantify a study from one acquisition manifest.
  • Add matched metagenomic or metatranscriptomic sequences when available.
  • Process large reservoirs in chunks when memory is limited.
  • Inspect peptide evidence, protein-group quantities and QC plots.
Input Supported route
Spectra One mzML or mzML.gz file per DDA acquisition
De novo predictions AlphaNovo CSV or generic sequence,score CSV, optionally gzipped
Protein sequences Prepared protein FASTA reservoir
Matched molecular data Optional specimen-matched DNA/RNA sequences and abundance tables

RAW conversion and de novo inference happen before FastaLake. See accepted inputs and preparing a study.

Installation

Route Best for
Docker A first run with the tools and examples already bundled
Source Python development or a Linux cluster
Apptainer / Slurm Running a study on a research cluster

With Docker installed:

docker pull --platform linux/amd64 ghcr.io/mannlabs/fasta-lake:v1.1.0rc2

The image includes Python, compiled Rust tools, Sage, directLFQ, AlphaPeptTools and example inputs. It targets Linux x86_64; Apple Silicon uses Docker's x86_64 emulation. The published release is v1.1.0rc2; development changes on main are available through the source or local Docker build instructions.

Quickstart

Run two small, real CAMPI acquisition windows and write the results locally:

mkdir -p fastalake-results
docker run --rm --platform linux/amd64 --network none \
  --memory 4g --memory-swap 4g --cpus 2 \
  --user "$(id -u):$(id -g)" -v "$PWD/fastalake-results:/work" \
  ghcr.io/mannlabs/fasta-lake:v1.1.0rc2 \
  fastalake-example campi --out /work/campi_01 --threads 2

Open these files in fastalake-results/campi_01/:

File Result
study/qc/QC_overview.png Quantities, group counts and completeness across acquisitions
study/study_group_matrix.tsv Protein-group quantities
study/peptide_evidence.tsv.gz Peptides supporting each group
workflow/samples/S01/inference/S01_razor.fasta The selected protein database

The example checks its outputs and saves VALIDATION.json. Use a new output folder for each run. Continue with your first study or reading the results.

Examples

Example What you learn
CAMPI Selection, searching, grouping and quantification
Chunked lookup Processing a reservoir within a memory budget
Matched DNA and RNA Adding molecular evidence to the selected database
Community analysis Exploring taxonomy and peptide–protein links

Browse the tutorials, follow the visual walkthrough, or use an assistant task file to work through a study.

Citation and credits

Treit et al. (2026). FastaLake selects compact metaproteomic databases from large sequence reservoirs by de novo peptide lookup. bioRxiv. Please also cite the software version used; see CITATION.cff. The manuscript analyses and source tables are maintained in fastalake-paper.

FastaLake uses Sage, directLFQ and AlphaPeptTools. Please cite these projects when using their results.

Contributing

Report a reproducible problem in Issues. For setup, tests and pull requests, see Contributing. Code is under the MIT licence; bundled data and dependencies retain their own terms.

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Protein database construction from de novo peptide evidence for proteomics and metaproteomics.

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