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

Xuejun Zhu; Peyton Shieh; Michael Su; Carolyn R. Bertozzi; Wenjun Zhang

doi:10.1039/C6CC05990B

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

Xuejun Zhu,^a Peyton Shieh,^b Michael Su,^a Carolyn R. Bertozzi^bc and Wenjun Zhang*^ad

Author affiliations

* Corresponding authors

^a Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94704, USA

^b Department of Chemistry, Stanford University, Stanford, CA 94305, USA

^c Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA

^d Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
E-mail: wjzhang@berkeley.edu

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.

This article is part of the themed collection: Bioorthogonal and click chemistry: Celebrating the 2022 Nobel Prize in Chemistry

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Article information

DOI: https://doi.org/10.1039/C6CC05990B
Article type: Communication
Submitted: 21 Jul 2016
Accepted: 19 Aug 2016
First published: 19 Aug 2016

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Chem. Commun., 2016,52, 11239-11242

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

X. Zhu, P. Shieh, M. Su, C. R. Bertozzi and W. Zhang, Chem. Commun., 2016, 52, 11239 DOI: 10.1039/C6CC05990B

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Chemical Communications

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

Abstract

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

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