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A fluorogenic screening platform enables directed evolution of an alkyne biosynthetic tool.

ABSTRACT: 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.

SUBMITTER: Zhu X

PROVIDER: S-EPMC5023497 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature

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A fluorogenic screening platform enables directed evolution of an alkyne biosynthetic tool.

Zhu Xuejun X Shieh Peyton P Su Michael M Bertozzi Carolyn R CR Zhang Wenjun W

Chemical communications (Cambridge, England) 20160825 75

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. ...[more]

PMID: 27561030

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