Mining the Arabidopsis genome for cytochrome P450 biocatalysts \n
Why this work is in the frame
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Bibliographic record
Abstract
Cytochromes P450 (CYPs) constitute a wide group of NAD(P)H-dependent monooxygenases, found throughout all kingdoms of life. Among the most important functions of CYPs are the synthesis of bioactive compounds and the conversion of xenobiotics. These functions can be translated into biotechnological applications, such as the production of highly regio- and stereo-specific drug metabolites for the pharmaceutical industry, or to confer activity towards toxic compounds for agronomics and bioremediation purposes. \nPlants possess a large number of CYP sequences, but most still remain uncharacterised, due to the difficulty in the isolation of the membrane-bound enzyme and in the reconstitution of an active and efficient redox system. \nIn this project, a fusion construct for the co-expression of the CYP with a suitable reductase was created. The construct consisted of a C-terminal Arabidopsis ATR2 reductase (codon-optimised for expression in E. coli and truncated of the N-terminal membrane anchor) connected through a poly-GlySer linker to the heme domain. An N-terminal Im9 peptide replaced the natural membrane-binding domain of the CYP. When CYP73A5 from Arabidopsis was cloned into the construct, it was able to convert almost 60 % of the substrate cinnamic acid to the hydroxylated derivative, in whole cell assays. This result demonstrated that this expression platform enables the expression of active redox self-sufficient P450 catalysts and it can be further utilised for the characterisation of orphan CYPs. \nFollowing from gene expression studies and reports on the existence of oxidative derivatives of TNT, the potential involvement of CYP81D11 in the detoxification of TNT was explored with different in planta assays, employing transgenic Arabidopsis lines and tobacco leaf discs. The results obtained were contrasting and did not provide a clear picture on the role of CYP81D11. Further studies have to be carried out in the future, using CYP81D11-knockout lines, as well as the purified enzyme.
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Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.001 | 0.001 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.002 | 0.000 |
| Research integrity | 0.001 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
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 it