A fluorogenic screening platform enables directed evolution of an alkyne biosynthetic tool†
Directed evolution was used to improve the activity of JamB, a membrane-bound bifunctional desaturase/acetylenase. To quickly assess the protein engineering outcomes, we developed a new platform for quantifying extracellular alkyne-tagged metabolites through a fluorogenic click reaction. Random mutagenesis yielded the best JamB variant with ∼20-fold increased activity in E. coli.
- This article is part of the themed collection: Bioorthogonal and click chemistry: Celebrating the 2022 Nobel Prize in Chemistry