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Selective Functionalization of Microgels with Enzymes by Sortagging.

Zhi Zou1,2, Elisabeth Gau1,3, Islam El-Awaad1,2,4

  • 1DWI - Leibniz-Institute for Interactive Materials , Forckenbeckstraβe 50 , 52074 Aachen , Germany.

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Summary

This study introduces sortagging, a selective enzyme immobilization method using microgels. This technique enhances enzyme stability and recyclability for industrial applications, demonstrated with five enzymes including a cost-effective cell lysate approach.

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Area of Science:

  • Biotechnology
  • Biocatalysis
  • Materials Science

Background:

  • Enzyme immobilization enhances enzyme stability and reusability in industrial processes.
  • Sortase-mediated enzyme immobilization offers a selective covalent attachment strategy.
  • Microgels provide a versatile platform for enzyme carrier development.

Purpose of the Study:

  • To investigate a sortase-mediated covalent immobilization strategy for enzymes onto microgels (GelZyms).
  • To assess the general applicability and selectivity of this immobilization technique.
  • To demonstrate the cost-effectiveness and application potential of the immobilized enzymes.

Main Methods:

  • Synthesis of aqueous poly(N-vinylcaprolactam)/glycidyl methacrylate (PVCL/GMA) microgels.
  • Tagging microgels with sortase A recognition peptide (LPETG) or nucleophilic tag (GGG).
  • Sortagging of five different enzymes (BSLA, Ym-phytase, CueO laccase, cellulase A2, P450 BM3), including immobilization from cell lysate.

Main Results:

  • Successful selective covalent immobilization of five distinct enzymes onto microgels.
  • All immobilized enzymes retained catalytic activity.
  • Enzymes demonstrated good recyclability, with >55% residual activity after six cycles for laccase CueO and P450 BM3 F87A.
  • Demonstrated application in dye bleaching using CueO-decorated microgels.

Conclusions:

  • Sortagging provides an effective and selective method for enzyme immobilization on microgels.
  • The developed GelZyms exhibit enhanced stability and recyclability, suitable for industrial biocatalysis.
  • Immobilization from cell lysate offers a cost-effective approach for enzyme applications.