Intensifying conformational dynamics enables HRP catalysis in organic phase

Area of Science:

• Biocatalysis
• Enzyme Engineering
• Organic Chemistry

Background:

• Organic solvents offer unique biocatalysis opportunities but often reduce enzyme activity.
• Horseradish peroxidase (HRP) typically shows decreased performance in organic media.

Purpose of the Study:

• To enhance HRP activity and substrate spectrum in pure toluene using polymer conjugation.
• To elucidate the mechanism behind improved enzyme performance in organic solvents.

Main Methods:

• Conjugation of HRP with Pluronic, an amphiphilic polymer.
• Assessing oxidative degradation of phenol/aniline derivatives in toluene.
• Low-field NMR spectroscopy and molecular dynamics simulations.

Main Results:

• Polymer-conjugated HRP achieved a substrate spectrum comparable to aqueous systems.
• Catalytic activity of conjugated HRP was significantly higher than free HRP in toluene.
• NMR and simulations revealed enhanced conformational dynamics and active site flexibility.

Conclusions:

• Amphiphilic polymer conjugation is a novel strategy to enhance enzyme activity in organic solvents.
• Improved enzyme flexibility and dynamics facilitate substrate uptake and catalysis.
• This approach advances biocatalysis in non-aqueous systems.