Ultra-high-throughput screening system for directed polymer binding peptide evolution

Lina Apitius, Kristin Rübsam, Christina Jakesch, Felix Jakob, Ulrich Schwaneberg
Biotechnology and Bioengineering 2019 April 14
Accumulation of plastics in the environment became a geological indicator of the Anthropocene era. An effective reduction of long-lasting plastics requires a treatment with microorganisms that release polymer-degrading enzymes. Polymer binding peptides function as adhesion promoters and enable a targeted binding of whole cells to polymer surfaces. An esterase A-based E. coli cell surface display screening system was developed, that enabled directed evolution of polymer binding peptides for improved binding strength to polymers. The E. coli cell surface screening system facilitates an enrichment of improved binding peptides from a culture broth through immobilization of whole cells on polymer beads. The polypropylene (PP)-binding peptide LCI was simultaneously saturated at five positions (Y29, D31, G35, E42, and D45; 3.2 million variants) and screened for improved PP-binding in the presence of the anionic surfactant sodium dodecylbenzenesulfonate (LAS; 0.25 mM). The cell surface system enabled efficient screening of the generated LCI diversity (in total approx. 10 million clones were screened). Characterization of identified LCI binders revealed an up to 12-fold improvement (eGFP-LCI-CSD-3: E42V/D45H) in PP-binding strength in the presence of the surfactant LAS (0.125 mM). The latter represents a first whole cell display screening system to improve adhesion peptides which can be used to direct and to immobilize organisms specifically to polymer surfaces (e.g. PP) and novel applications (e.g. in targeted plastic degradation). This article is protected by copyright. All rights reserved.

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