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Ultrahigh-throughput screening system for directed polymer binding peptide evolution.

Lina Apitius1,2, Kristin Rübsam1, Christina Jakesch2

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

Biotechnology and Bioengineering
|April 16, 2019
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Summary
This summary is machine-generated.

Researchers developed a novel Escherichia coli cell surface display system to enhance polymer-binding peptides. This system improves targeted plastic degradation by enabling microorganisms to bind more strongly to plastic surfaces, even in the presence of surfactants.

Keywords:
E. coli cell surface displayanchor peptidesand polypropylenedirected evolutionultrahigh throughput

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Area of Science:

  • Biotechnology and Environmental Science
  • Microbial Engineering for Plastic Remediation

Background:

  • Environmental plastic accumulation is a significant global challenge, necessitating innovative degradation strategies.
  • Microbial enzymes offer a promising avenue for breaking down persistent polymers.
  • Targeted cell adhesion to plastic surfaces is crucial for efficient microbial degradation.

Purpose of the Study:

  • To develop a whole-cell display screening system for directed evolution of polymer-binding peptides.
  • To enhance the binding affinity of peptides to polypropylene (PP) in the presence of surfactants.
  • To create a platform for immobilizing microorganisms onto plastic surfaces for targeted applications.

Main Methods:

  • An esterase A-based Escherichia coli cell surface display system was engineered.
  • Directed evolution was employed to screen millions of polymer-binding peptide variants (LCI) for improved PP binding.
  • Enrichment of high-affinity binders was achieved through immobilization on polymer beads in the presence of sodium dodecylbenzenesulfonate (LAS).

Main Results:

  • The screening system successfully identified improved polypropylene-binding peptides from a large library (~10 million clones).
  • Mutations (E42V/D45H) in the LCI peptide resulted in up to a 12-fold increase in PP-binding strength.
  • Enhanced binding was observed even in the presence of the surfactant LAS.

Conclusions:

  • This study presents the first whole-cell display screening system for improving adhesion peptides targeting polymer surfaces.
  • The developed system enables efficient directed evolution and immobilization of microorganisms onto plastics like PP.
  • This technology holds potential for novel applications in targeted plastic degradation and bioremediation.