Additional file 1 of Reduction of phenolics in faba bean meal using recombinantly produced and purified Bacillus ligniniphilus catechol 2,3-dioxygenase
Bibliographic record
Abstract
Additional file 1: Figure S1. Purification of BLC23O by size-exclusion chromatography. A) Size-exclusion chromatography (SEC) was performed with a HiLoad16/600 Superdex200 column attached to an FPLC system and run at a flowrate of 1 mL/min. A) The elution profile (OD280nm) with the ladder at the bottom of the graph representing the fractions collected during the run. B)Elution profile for standard proteins to calibrate the SEC column where F is ferritin, Ald is aldolase, C is conalbumin, O is ovalbumin (45kDa), CA is carbonic anhydrase (29.2 kDa), R is RNase A, and Apr is aprotinin (Cytvia). C)A 12% acrylamide SDS-PAGE analysis of select fractions from the BLC23O SEC purification. Figure S2. Folin–Ciocalteu assay development. A) Calibration curve for determination of total phenolic content. Folin–Ciocalteu reagent was added to known concentrations of tannic acid ranging from 0 to 30 ug/mL and left to react for 45 min. Absorbances were then measured at 725 nm. The data represent the mean and standard deviation (n=3). A linear fit of the data with an equation of y=0.0127x and a R2 value of 0.9981 are shown. B) Evaluation of Folin–Ciocalteu reagent kinetics. Reaction solutions with volumes of 250 μL were prepared with 0.125 N Folin–Ciocalteu reagent, 0.125 mg/mL sodium carbonate, and 14 μg/mL tannic acid. The absorbance at 725 nm was monitored for 180 min in 15-min intervals. The data represent the mean and standard deviation (n=3). Figure S3. Representative images of commercially available phenolic compounds tested as substrates for BLC23O. Figure S4. Biocatalytic reduction of phenols in purified phenolic compounds using BLC23O. Top row) Phenolic content in samples containing purified compounds as indicated, was evaluated in the presence (black) and absence (blue) of BLC23O over time. An enzyme-only control sample was also assessed (grey). Reactions were initiated with the addition enzyme at 32.5 °C. Samples were taken in 30 min intervals and reacted with the Folin–Ciocalteu reagent. Time zero was set to 100 % phenol and changes in absorbance at 725nm plotted on a percentage change basis. Bottom row) The observed changes in phenol content of BLC23O treated compounds as indicated were quantified relative to untreated fractions. The data represent the mean and standard deviation (n=3). Linear fits of the data and the corresponding equations and coefficients of determination are shown.
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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 | 0.002 | 0.017 |
| Meta-epidemiology (narrow) | 0.002 | 0.001 |
| Meta-epidemiology (broad) | 0.002 | 0.001 |
| Bibliometrics | 0.002 | 0.004 |
| Science and technology studies | 0.001 | 0.000 |
| Scholarly communication | 0.003 | 0.002 |
| Open science | 0.002 | 0.001 |
| Research integrity | 0.002 | 0.001 |
| Insufficient payload (model declined to judge) | 0.882 | 0.199 |
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".