Production of <scp>C6–C14</scp> Medium‐Chain Fatty Acids in Seeds and Leaves <i>via</i> Overexpression of Single Hotdog‐Fold Acyl‐Lipid Thioesterases
Bibliographic record
Abstract
ACYL-LIPID THIOESTERASES (ALT) are a type of plant acyl-acyl carrier protein thioesterase that generate a wide range of medium-chain fatty acids and methylketone (MK) precursors when expressed heterologously in Escherichia coli. While this makes ALT-type thioesterases attractive as metabolic engineering targets to increase production of high-value medium-chain fatty acids and MKs in plant systems, the behavior of ALT enzymes in planta was not well understood before this study. To profile the substrate specificities of ALT-type thioesterases in different plant tissue types, AtALT1-4 from Arabidopsis thaliana, which have widely varied chain length and oxidation state preferences in E. coli, were overexpressed in Arabidopsis seeds, Camelina sativa seeds, and Nicotiana benthamiana leaves. Seed-specific overexpression of ALT enzymes led to medium-chain fatty acid accumulation in Arabidopsis and Camelina seed triacylglycerols, and transient overexpression in N. benthamiana demonstrated that the substrate preferences of ALT-type thioesterases in planta generally agree with those previously determined in E. coli. AtALT1 and AtALT4 overexpression in leaves and seeds resulted in the accumulation of 12-14 carbon-length fatty acids and 6-8 carbon-length fatty acids, respectively. While it was difficult to completely profile the products of ALT-type thioesterases that generate MK precursors (i.e. β-keto fatty acids), our results nonetheless demonstrate that ALT enzymes are catalytically diverse in planta. The knowledge gained from this study is a significant step towards being able to use ALT-type thioesterases as metabolic engineering tools to modify the fatty acid profiles of oilseed crops, other plants, and microorganisms.
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How this classification was reachedexpand
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 | 0.001 |
| 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 teacher head, 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".