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Stabilization of interface-binding chloroperoxidase for interfacial biotransformation.

Ravindrabharathi Narayanan1, Guangyu Zhu, Ping Wang

  • 1Department of Chemical Engineering, The University of Akron, Akron, OH 44325-3906, USA.

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Poly(ethylene glycol) (PEG) significantly enhances chloroperoxidase (CPO) stability and productivity by protecting against hydrogen peroxide (H2O2) deactivation. Stabilizers also improved CPO enantioselectivity, offering new avenues for enzyme applications.

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Area of Science:

  • Biocatalysis
  • Enzyme Stabilization
  • Interface Chemistry

Background:

  • Chloroperoxidase (CPO) is crucial for biocatalysis but susceptible to deactivation by hydrogen peroxide (H2O2).
  • Interface-assembled CPO exhibits enhanced stability, yet H2O2-induced deactivation remains a limiting factor for enzyme productivity.

Purpose of the Study:

  • To investigate methods for improving the operational and storage stability of interface-bound CPO.
  • To evaluate the efficacy of various stabilizers and in situ H2O2 generation on CPO performance.

Main Methods:

  • Conjugating native CPO with polystyrene for self-assembly at a water-oil interface.
  • Testing stabilizers like poly(ethylene glycol) (PEG), poly(ethyleneimine) (PEI), glycerol, sugars, and sucrose monododecanoate.
  • Employing in situ H2O2 generation using glucose oxidase (GOx).

Main Results:

  • PEG significantly enhanced both operational and storage stability, increasing CPO productivity by up to 57%.
  • PEI improved storage stability but not operational stability, while glucose enhanced operational stability twofold.
  • In situ H2O2 generation by GOx extended operational lifetime by combining glucose's stabilizing effect and lower H2O2 concentrations.
  • Stabilizers improved CPO enantioselectivity by up to 10%.

Conclusions:

  • PEG is a highly effective stabilizer for interface-bound CPO, boosting productivity and stability.
  • Stabilizers offer a promising strategy to overcome H2O2-induced deactivation and enhance CPO enantioselectivity.
  • In situ H2O2 generation provides a viable method to prolong CPO operational lifetime in biocatalytic processes.