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

You do not have JavaScript enabled. Please enable JavaScript to access the full features of the site or access our non-JavaScript page.

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

Download options Please wait...

Supplementary files

Supplementary information PDF (999K)

Article information

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

Download Citation

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

Permissions

Request permissions

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

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Social activity

Fetching data from CrossRef.
This may take some time to load.

Chemical Communications

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

Abstract

Supplementary files

Article information

Download Citation

Permissions

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

Social activity

Search articles by author

Spotlight

Advertisements