comp-001 — Uricase Shio-Koji Protease Stability¶
Question: Will A. flavus uricase (Q00511) survive the protease environment of a shio-koji ferment (15–20% NaCl, pH 4.5–5.0, RT, 7–14 days) with meaningful activity retained?
Short answer: Computational analysis predicts low risk. All protease recognition sites are in confidently-folded regions (100% of residues pLDDT > 80). Combined with strong salt inhibition of the dominant protease (ALP retains ~19% activity at shio-koji NaCl concentrations), the structural evidence argues against significant proteolytic degradation.
Verdict: §1.10 wet-lab stability experiment should still run — this analysis shifts the prior from "unknown" to "probably fine," reframing §1.10 as confirmation rather than feasibility gate.
How to reproduce¶
Requirements: Python 3.8+, stdlib only (json, pathlib, os). No packages to install.
Re-running with updated data¶
- New AlphaFold model: Download updated PDB from
https://alphafold.ebi.ac.uk/files/AF-Q00511-F1-model_v<version>.pdb, re-extract pLDDT values intoinputs/alphafold_Q00511_plddt.json, updateinputs/provenance.md, re-run. - Updated protease specificities: Edit
inputs/protease_specificities.jsonand update_sourcefield with new MEROPS reference. - Different protein: Replace
Q00511.fastaandalphafold_Q00511_plddt.jsonwith inputs for the new target.
File index¶
inputs/
Q00511.fasta UniProt sequence (fetched 2026-05-05)
alphafold_Q00511_plddt.json Per-residue pLDDT from AlphaFold v6 PDB (fetched 2026-05-05)
protease_specificities.json P1/P1' rules + salt inhibition data (MEROPS + primary literature)
provenance.md Data sources, fetch dates, version notes
outputs/
cleavage_sites.json Full machine-readable results
summary.md Human-readable findings — this is the artifact cited in the wiki
analyze.py Analysis script
README.md This file
Wiki links¶
- Interpretive page:
wiki/uricase-protease-stability-computational.md - Tracking index:
wiki/computational-experiments.md - Wet-lab experiment this informs:
wiki/validation-experiments.md§1.10 - Platform context:
wiki/koji-endgame-strain.md