Multifactorial interaction and influence of culture conditions on yellow fluorescent protein production in Phaeodactylum tricornutum
Bibliographic record
Abstract
• Bioengineered P. tricornutum expressing yellow fluorescent protein extrachromosomally. • A model was developed to predict culture parameters on YFP production in P. tricornutum. • Optimized conditions increased YFP fluorescence and yield by 4.2x and 1.8x, respectively. • Inhibition of the ubiquitin–proteasome system increased YFP levels by 50–150 % after 4–8 h. • Cycloheximide treatment significantly reduced YFP stability at 20 h. Phaeodactylum tricornutum is a promising host for light-driven synthesis of heterologous proteins. However, the marine cold-water environment and alkaline-acidic pH shifts in the culture, necessitated by the diatom’s growth requirements. In this study, we analyzed the influence of growth condition on maturation and dynamics of the yellow fluorescent protein (YFP) in episomal-transformant P. tricornutum . A mathematical model was developed to detect the parameters that affect biomass and YFP production. Optimized conditions increased YFP mean fluorescence intensity (MFI) per cell by 4.2-fold (3.6 ± 0.6 to 15.4 ± 1.1) and total protein levels in the culture by 1.8-fold (123 ± 4 to 219 ± 9 µg L −1 ), without affecting biomass. YFP stability studies in P. tricornutum showed that the ubiquitin–proteasome system contributes the degradation of the recombinant protein, whereas newly synthesized YFP remains stable for up to 12 h. This optimization provides insights into the fluorescent protein-based heterologous production in diatoms.
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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 | 0.001 | 0.001 |
| 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".