Display of large enzyme complexes on the cell surface of <i>Bacillus subtilis</i>
Bibliographic record
Abstract
The ability to display multi‐subunit enzyme complexes on the bacterial cell surface offers opportunities to degrade complex polymers such as plant cellulose in an efficient and cooperative manner. To develop an engineered nanomachinery for surface display of enzymes, a fusion protein consists of streptavidin followed by a flexible linker and a cell wall binding domain derived from Bacillus subtilis cell wall hydrolase, LytE, is designed. Since each streptavidin subunit can self‐assemble into the tetrameric state and capture an enzyme (β‐lactamase in this study) carrying a streptavidin binding tag (SBP), a 140‐kDa complex is expected to form. Also, the tetrameric streptavidin fusion can potentially enhance the cell wall binding avidity. To examine the display efficiency, linkers with different lengths (51 and 151 amino acids) in the streptavidin fusions are designed. SDS‐PAGE results showed that the streptavidin fusions could be displayed as tetramers on the cell surface. Enzyme assay and Western blot showed that SBP‐tagged β‐lactamase could be captured to the cell surface. Interestingly, cells displaying streptavidin fusions with a longer linker showed 44% higher β‐lactamase activity. Longer linker is suggested to offer better surface accessibility of the displayed streptavidin fusions. This system can potentially be applied to display one or multiple species of SBP‐tagged proteins on the cell surface.
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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 | 0.000 |
| 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 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".