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Record W4394011973 · doi:10.1101/2024.04.05.588253

Genome mining leads to the identification of a stable and promiscuous Baeyer-Villiger monooxygenase from a thermophilic microorganism

2024· preprint· en· W4394011973 on OpenAlexafffund
Amir R. Bunyat‐zada, Stephan E. Ducharme, Maria E. Cleveland, Esther R. Hoffman, Graeme W. Howe

Bibliographic record

VenuebioRxiv (Cold Spring Harbor Laboratory) · 2024
Typepreprint
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicEnzyme Catalysis and Immobilization
Canadian institutionsQueen's University
FundersNatural Sciences and Engineering Research Council of Canada
KeywordsThermophileMicroorganismMonooxygenaseIdentification (biology)GenomeBiologyComputational biologyGeneticsGeneEcologyEnzymeBiochemistryBacteriaCytochrome P450

Abstract

fetched live from OpenAlex

Abstract Baeyer-Villiger monooxygenases are NAD(P)H-dependent flavoproteins that catalyze oxygen insertion reactions which convert ketones to valuable esters and lactones. While these enzymes offer an appealing alternative to traditional Baeyer-Villiger oxidations, these proteins tend to be either too unstable or exhibit too narrow of a substrate scope for implementation as industrial biocatalysts. Here, sequence similarity networks were used to search for novel Baeyer-Villiger monooxygenases that are both stable and substrate promiscuous. Our genome mining efforts led to the identification of an enzyme from Chloroflexota bacterium (strain G233) dubbed ssn BVMO that exhibits i) the highest melting temperature recorded to date for a naturally sourced Baeyer-Villiger monooxygenase, ii) a remarkable kinetic stability across a wide range of conditions, and iii) a broad substrate scope that includes linear aliphatic, aromatic, and sterically bulky ketones. Kinetic characterization of this enzyme was undertaken to identify the optimal conditions for ssn BVMO catalysis, and a subsequent quantitative assay using propiophenone as a substrate afforded more than 95% conversion. To spur the implementation of this enzyme as an oxidative biocatalyst, several fusion proteins were constructed that linked ssn BVMO to a thermostable phosphite dehydrogenase. These self-sufficient enzymes can recycle NADPH and permit oxidations to be run with sub-stoichiometric quantities of this expensive cofactor. Extensive characterization of these fusion enzymes permitted identification of PTDH-L1- ssn BVMO as the most promising oxidative biocatalyst. Results described herein demonstrate that this new monooxygenase has significant potential as a useful industrial biocatalyst for Baeyer-Villiger oxidations.

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 score0.021
Threshold uncertainty score1.000

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.001
Research integrity0.0000.000
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 head0.008
GPT teacher head0.209
Teacher spread0.201 · 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

Citations0
Published2024
Admission routes2
Has abstractyes

Explore more

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