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Record W4400386925 · doi:10.1101/2024.07.04.602010

Discovery of the missing cytochrome P450 monooxygenase cyclases that conclude glyceollin biosynthesis in soybean

2024· preprint· en· W4400386925 on OpenAlexaff
Praveen Khatri, Kuflom Kuflu, Tim McDowell, Jie Lin, Nikola Kovinich, Sangeeta Dhaubhadel

Bibliographic record

VenuebioRxiv (Cold Spring Harbor Laboratory) · 2024
Typepreprint
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicPlant Gene Expression Analysis
Canadian institutionsYork UniversityAgriculture and Agri-Food CanadaWestern University
Fundersnot available
KeywordsMonooxygenaseBiochemistryBiosynthesisEnzymeCytochrome P450BiologyHeterologous expressionTranscriptomePhytophthora sojaeIsoflavonoidPrenyltransferaseGeneGene expressionFlavonoidRecombinant DNA

Abstract

fetched live from OpenAlex

Abstract Glyceollins are isoflavonoid-derived metabolites produced by soybean that hold great promise in improving human and animal health due to their antimicrobial, and other medicinal properties. They play important roles in agriculture by defending soybean against one of its most destructive pathogens, Phytophthora sojae . Longstanding research efforts have focused on improving accessibility to glyceollins, yet chemical synthesis remains uneconomical. The fact that some of the key genes involved in the final step of glyceollin biosynthesis have not been identified, engineering the accumulation of these important compounds in microbes is not yet possible. Although the activity of a P450 cyclase was inferred to catalyze the final committed step in glyceollin biosynthesis forty years ago, the enzyme in question has never been conclusively identified. This study reports, for the first time, the identification of three cytochrome P450 monooxygenase cyclases that catalyze the final steps of glyceollin biosynthesis. Utilizing P. sojae -soybean transcriptome data, along with genome mining tools and co-expression network analysis, we have identified 16 candidate glyceollin synthases (GmGS). Heterologous expression of these candidate genes in yeast, coupled with in vitro enzyme assays, enabled us to discover three enzymes capable of producing two glyceollin isomers. GmGS11A and GmGS11B catalyzed the conversion of glyceollidin to glyceollin I, whereas GmGS13A converted glyceocarpin to glyceollin III. The functionality of these candidates was further confirmed in planta through gene silencing and overexpression in soybean hairy roots. This groundbreaking study not only contributes to the understanding of glyceollin biosynthesis, but also demonstrates a new synthetic biology strategy that could potentially be scaled up to produce valuable molecules for crop and disease management.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame distilled prediction

Teacher imitation

Not 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.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: codex-gemma-dda1882f352aValidation status: machine_predicted_unvalidated
Candidate categoriesMeta-epidemiology (narrow)
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.011
Threshold uncertainty score1.000

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0010.000
Meta-epidemiology (broad)0.0010.001
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0010.002
Research integrity0.0010.001
Insufficient payload (model declined to judge)0.0000.000

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.

Opus teacher head0.012
GPT teacher head0.225
Teacher spread0.213 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one teacher head, not a consensus.

Study designBench or experimental
Domainnot available
GenreEmpirical

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".

Quick stats

Citations3
Published2024
Admission routes1
Has abstractyes

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