A fluorogenic screening platform enables directed evolution of an alkyne biosynthetic tool
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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.
author list (cited authors)
Zhu, X., Shieh, P., Su, M., Bertozzi, C. R., & Zhang, W.