The epoxide hydrolase from Alphaproteobacteria bacterium was identified to convert meso-3,4-epoxytetrahydrofuran (1) to (3R,4R)-tetrahydrofurandiol [(RR)-2] predominantly. The enzyme was engineered via directed evolution to revert its original enantioselectivity to generate (3S,4S)-tetrahydrofurandiol [(SS)-2] in quantitative yield and to tolerate an extremely high substrate concentration desired for industrial processes.

Abstract

Chiral vicinal diols are important intermediates in the synthesis of pharmaceuticals. Epoxide hydrolases catalyze hydrolytic ring opening of epoxides to produce the corresponding vicinal diols, providing an attractive way to access these building blocks under mild conditions in a stereoselective and atom-efficient manner. In this study, an epoxide hydrolase is identified and engineered to form (3S,4S)-tetrahydrofurandiol in high optical purity via the desymmetrization of meso-3,4-epoxytetrahydrofuran. In nine rounds of directed evolution, the enzyme's native (3R,4R)-stereopreference was reversed and its activity was dramatically improved to achieve quantitative yield under remarkably high 500 g/L substrate concentration and low enzyme loading. Computational modelling provides insights on the changes in enzyme-substrate interaction that result in divergent enantioselectivities afforded by evolved variants.

Conflict of interest

The authors declare no conflict of interest.

Open Research

Data Availability Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Supporting Information

References

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Volume15, Issue12

June 22, 2023

e202300238

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Catalysts in Industry

Directed Evolution to Reverse Epoxide Hydrolase Enantioselectivity for meso-3,4-Epoxytetrahydrofuran

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Abstract

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Data Availability Statement

Supporting Information

References

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Directed Evolution to Reverse Epoxide Hydrolase Enantioselectivity for meso-3,4-Epoxytetrahydrofuran

Graphical Abstract

Abstract

Conflict of interest

Open Research

Data Availability Statement

Supporting Information

References

References

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