Engineering a Carbonyl Reductase for High-Efficiency Synthesis of Optically Pure (S)-Pro-Xylane: An Alternative...

Area of Science:

• Biocatalysis
• Enzyme Engineering
• Cosmetic Chemistry

Background:

• Pro-Xylane is a key C-glycoside in cosmetics.
• Conventional synthesis yields byproducts and favors less active isomers.
• Enzymatic reduction offers a stereoselective and sustainable alternative.

Purpose of the Study:

• To engineer carbonyl reductase (CR) for efficient production of (S)-Pro-Xylane.
• To improve the stereoselectivity and yield of the reduction step.
• To develop a sustainable alternative to chemical synthesis.

Main Methods:

• Site-directed mutagenesis of carbonyl reductase (CoCR13) from Candida orthopsilosis.
• Enzymatic reduction of 1-C-(β-d-xylopyranosyl)-acetone.
• Kinetic analysis (kcat/Km) and diastereomeric excess determination.
• Molecular dynamics simulations.

Main Results:

• Engineered variant R129E/D210F showed a 245.33-fold increase in kcat/Km over wild-type.
• Achieved >99% (β, S) diastereomeric excess.
• Reached a spatiotemporal yield of 49.97 g·L-1·h-1 with 1300 mM substrate.
• Simulations revealed altered substrate binding and tunnel dynamics.

Conclusions:

• Engineered CR provides a highly efficient and stereoselective route to (S)-Pro-Xylane.
• This biocatalytic approach offers a sustainable and environmentally friendly production method.
• Expands the application of CR in the synthesis of valuable cosmetic ingredients.