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Stereoselective C-C bond formation catalysed by engineered carboxymethylproline synthases
Refaat B Hamed1, J Ruben Gomez-Castellanos, Armin Thalhammer
1Department of Chemistry, Chemistry Research Laboratory, University of Oxford, Mansfield Road, Oxford OX1 3TA, UK.
Protein engineering enables crotonase superfamily enzymes to catalyze stereoselective C-C bond formation for N-heterocycles. This advances synthetic organic chemistry using biocatalysis for enolate reactions.
Area of Science:
- Organic Chemistry
- Biocatalysis
- Protein Engineering
Background:
- Enolate chemistry is crucial for stereoselective C-C bond formation in organic synthesis.
- Protein-based catalysts have seen limited success in controlling enolate reactions for C-C bond formation.
Purpose of the Study:
- To engineer 5-carboxymethylproline synthases for stereoselective C-C bond formation.
- To enable the synthesis of N-heterocycles using controlled enolate intermediates.
Main Methods:
- Protein engineering of 5-carboxymethylproline synthases (crotonase superfamily).
- Introduction of active site substitutions, including at the oxyanion binding site.
- Analysis of stereochemical outcomes in C-C bond formation reactions.
Main Results:
- Engineered enzymes produced substituted N-heterocycles with high diastereomeric excesses.
- Demonstrated stereocontrolled enolate formation and reaction.
- Highlighted the adaptability of the crotonase superfamily for enolate chemistry.
Conclusions:
- The crotonase superfamily can be adapted as versatile catalysts for stereoselective enolate chemistry.
- Protein engineering offers a powerful approach to control C-C bond formation via enolates.
- This work expands the utility of biocatalysis in synthesizing complex N-heterocycles.
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