Chalcone isomerase-like (CHIL) impedes the lactone shunt and modulates flux partitioning in isoflavonoid biosynthesis
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Bibliographic record
Abstract
ABSTRACT The reconstitution of biosynthetic pathways in heterologous hosts is often challenged by the transition to a foreign cellular environment, lacking compatible structural or regulatory features. Auxiliary or non-catalytic proteins can play a critical role in guiding metabolic flux. Chalcone isomerase-like (CHIL) is a non-catalytic protein known to serve as a partner to chalcone synthase (CHS) in flavonoid biosynthesis, rectifying its promiscuous activity and preventing the formation of by-products, such as p -coumaroyltriacetic acid lactone (CTAL). Here, we extended the functional analysis of CHILs to the legume-characteristic isoflavonoid pathway. We assessed CHIL orthologs, using sequence analysis and structural modelling to predict their respective binding capacities, followed by in vitro characterization. The addition of CHIL to enzyme assays containing CHS, alone or with downstream enzymes, chalcone reductase (CHR) and chalcone isomerase (CHI), reduced CTAL levels (up to 42%) while simultaneously increasing the output of desired intermediates. Combinatorial yeast biotransformation assays described a critical role for CHIL in conducting flux through chalcone, flavanone, and isoflavone biosynthesis. The inclusion of CHIL in engineered yeast strains enhanced overall titers and, unexpectedly, promoted partitioning toward the deoxy-branch (isoliquiritigenin, liquiritigenin, and daidzein) up to 67%, with a 33% increase in final daidzein titers. Therefore, we have revealed an expanded role for CHIL as an auxiliary protein in isoflavonoid biosynthesis and underscored the utility of non-catalytic proteins in metabolic engineering.
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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 it