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Enzymatic cascade reaction in simple-coacervates.

Ritu Toor1, Lysandre Hourdin1, Sharvina Shanmugathasan1

  • 1Univ. Bordeaux, CNRS, Bordeaux INP, ISM, UMR 5255, Site ENSCBP, 16 avenue Pey Berland, 33607 Pessac, France.

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|September 24, 2022
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Enzymatic cascade reactions were successfully demonstrated in coacervate droplets, acting as mini-reactors. These all-aqueous droplets preserve enzyme integrity and activity, offering insights into proximity and confinement effects.

Keywords:
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Area of Science:

  • Biotechnology
  • Chemical Engineering
  • Biochemistry

Background:

  • Enzymatic droplet-sized reactors have broad applications in diagnostics, purification, and industry.
  • Coacervates offer a simple, all-aqueous model for studying enzyme cascade reactions, preserving enzyme integrity.
  • The impact of enzyme sequestration and proximity within coacervates on their activity requires further investigation.

Purpose of the Study:

  • To construct and investigate enzymatic reactors using coacervates of ampholyte polymer chains stabilized with agar.
  • To demonstrate the ability of these coacervates to sequester enzymes like glucose oxidase and catalase while maintaining their catalytic activity.
  • To explore the influence of proximity and confinement within coacervates on enzymatic cascade reactions.

Main Methods:

  • Construction of enzymatic reactors utilizing simple coacervation of ampholyte polymer chains stabilized with agar.
  • Demonstration of enzyme sequestration (glucose oxidase, catalase) and activity preservation within coacervates.
  • Analysis of enzymatic cascade reactions via colorimetric detection of resazurin reduction to resorufin and its regeneration.

Main Results:

  • Coacervates successfully sequestered glucose oxidase and catalase, preserving their catalytic activity.
  • An enzymatic cascade reaction involving glucose oxidase and catalase was demonstrated within the coacervates.
  • The colorimetric assay using resazurin reduction showed a clear indication of the enzymatic cascade, with resorufin regeneration observed.

Conclusions:

  • Coacervates serve as effective enzymatic reactors, capable of sequestering enzymes and facilitating cascade reactions.
  • The proximity and confinement of enzymes within coacervates do not impede, and may influence, their catalytic activity.
  • This study provides valuable insights into the behavior of enzymes in confined, all-aqueous environments for potential biotechnological applications.