γ-Glutamyl Cysteine Ligase of <i>Lactobacillus reuteri</i> Synthesizes γ-Glutamyl Dipeptides in Sourdough
Bibliographic record
Abstract
Kokumi-active γ-glutamyl dipeptides (γ-GPs) accumulate in fermented food. γ-Glutamyl transferase, glutaminase, glutathione synthetase, and γ-glutamyl cysteine ligase (GCL) may synthesize γ-GPs. The genome of Lactobacillus reuteri encodes GCL but not glutathione synthetase or glutamyl transferase; therefore, this study investigated the role of GCL in γ-GP synthesis by L. reuteri LTH5448. Phylogenomic analysis of gcl in lactobacilli demonstrated that three genes coding for GCL are present in L. reuteri; two of these are present in L. reuteri LTH5448. Two deletion mutants of L. reuteri LTH5448, L. reuteri LTH5448Δ gcl1 and LTH5448Δ gcl1Δ gcl2, were constructed by double crossover mutagenesis. Growth and oxygen resistance of the mutants were comparable to the wild type. γ-Glu-Glu, γ-Glu-Leu, γ-Glu-Ile, γ-Glu-Val, and γ-Glu-Cys were quantified in buffer and sourdough fermentations by liquid chromatography-mass spectrometry. The wild type and L. reuteri Δ gcl1 but not Δ gcl1Δ gcl2 converted amino acids to γ-Glu-Cys. γ-Glu-Ile accumulation was reduced in both mutants; however, the disruption of gcl did not alter the biosynthesis of the other γ-GPs. In conclusion, gcl1 in L. reuteri mediates γ-Glu-Ile synthesis, gcl2 mediates γ-Glu-Cys synthesis, but neither gene affected synthesis of other γ-GPs. This study facilitates selection of starter cultures that synthesize γ-Glu peptides with kokumi activity and, thus, improve the taste of fermented foods.
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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) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 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".