Abstracts presented at the 37th Annual Meeting of The Protein Society. ABS#599: Catechol 1,2-dioxygenase with improved thermostability
Bibliographic record
Abstract
Catechol 1,2-dioxygenase (C12O) is involved in the catabolism of aromatic compounds and biosynthesis of cis, cis-muconic acid, an important precursor chemical and a compound of potential relevance to petrochemical replacement. However, the low thermostability of C12O is a hindrance to industrial application performance. In this study, the thermostability of a mesophilic C12O from Rhodococcus opacus was enhanced by the rational design and engineering of disulphide bonds in the protein structure. A computational pipeline of existing algorithms was used to identify possible locations for the introduction of stabilizing disulphide bonds. A set of selection criteria were used to narrow down the list of candidates, and molecular dynamic simulations were performed for each of the selected candidates under simulated thermal denaturing conditions. Subsequent wet bench testing of the selected candidates showed that two disulfide locations in particular (including an intra-subunit bond and an inter-subunit bond) yielded significant increases to protein thermostability. These two mutants showed greater than 5°C rises in melting temperature and T50, as well as significant increases in half-life at 40°C. Current work on a double mutant of these is ongoing to determine whether the effects of increased stability are additive. Overall, we conclude that the introduction of disulfide bonds resulted in significantly improved thermostability of C12O compared to wild type.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.001 |
| Meta-epidemiology (narrow) | 0.002 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.001 | 0.001 |
| Scholarly communication | 0.002 | 0.001 |
| Open science | 0.001 | 0.001 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.200 | 0.103 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".