Isolation and characterization of a Δ5 FA desaturase from <i>Pythium irregulare</i> by heterologous expression in <i>Saccharomyces cerevisiae</i> and oilseed crops
Bibliographic record
Abstract
By using the polymerase chain reaction approach with two degenerate primers targeting the heme-binding and the third histidine-rich motifs in microsomal carboxyl-directed desaturases, we identified a cDNA PiD5 from Pythium irregulare encoding a delta5 desaturase. The substrate specificity of the enzyme was studied in detail by expressing PiD5 in a yeast (Saccharomyces cerevisiae) mutant strain, AMY-2alpha, where ole1, a delta9 desaturase gene, is disrupted. The result revealed that the encoded enzyme could desaturate unsaturated FA from 16 to 20 carbons beginning with delta9 and delta11 as well as delta8 ethylenic double bonds. Introduction of PiD5 into Brassica juncea under the control of a CaMV 35S constitutive promoter resulted in accumulation of several delta5-unsaturated polymethylene-interrupted FA (delta5-UPIFA) including 18:2-5,9, 18:2-5,11, 18:3-5,9,12, and 18:4-5,9,12,15 in vegetative tissues. The transgenic enzyme could also desaturate the exogenously supplied homo-gamma-linolenic acid (20:3-8,11,14) to arachidonic acid (20:4-5,8,11,14). Introduction of PiD5 into B. juncea and flax under the control of seed-specific promoters resulted in production of delta5-UPIFA, representing more than 10% of the total FA in the seeds.
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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 | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 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 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".