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Updated: Jul 11, 2025

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening
Published on: April 1, 2016
Secretion and directed evolution of unspecific peroxygenases in S. cerevisiae
Niklas Dietz1, Li Wan1, Judith Münch1
1Institute of Chemistry, Martin-Luther-University Halle-Wittenberg Weinbergweg 22, Halle (Saale), Germany.
Abstract:
Yeast-based secretion systems are advantageous for engineering highly interesting enzymes that are not or barely producible in E. coli. The herein-presented production setup facilitates high-throughput screening as no cell lysis is required. All techniques are described in detail, with access to freely available online tools and all vectors have been made available on the non-profit plasmid repository AddGene. We describe the method for UPOs as a model enzyme, showcasing their secretion, detection, and evolution using S. cerevisiae. Additional material to transfer this to P. pastoris has been published by our group previously (Püllmann & Weissenborn, 2021).

