Format

Send to

Choose Destination
Methods Mol Biol. 2018;1671:269-290. doi: 10.1007/978-1-4939-7295-1_17.

Design, Engineering, and Characterization of Prokaryotic Ligand-Binding Transcriptional Activators as Biosensors in Yeast.

Author information

1
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark.
2
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark. mije@biosustain.dtu.dk.
3
Joint BioEnergy Institute, Emeryville, CA, USA.
4
Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
5
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA, USA.
6
Department of Bioengineering, University of California, Berkeley, CA, USA.

Abstract

In cell factory development, screening procedures, often relying on low-throughput analytical methods, are lagging far behind diversity generation methods. This renders the identification and selection of the best cell factory designs tiresome and costly, conclusively hindering the manufacturing process. In the yeast Saccharomyces cerevisiae, implementation of allosterically regulated transcription factors from prokaryotes as metabolite biosensors has proven a valuable strategy to alleviate this screening bottleneck. Here, we present a protocol to select and incorporate prokaryotic transcriptional activators as metabolite biosensors in S. cerevisiae. As an example, we outline the engineering and characterization of the LysR-type transcriptional regulator (LTTR) family member BenM from Acetinobacter sp. ADP1 for monitoring accumulation of cis,cis-muconic acid, a bioplast precursor, in yeast by means of flow cytometry.

KEYWORDS:

Biosensor; Cell factory; Screening; Synthetic biology; Transcription factor; Yeast

PMID:
29170965
DOI:
10.1007/978-1-4939-7295-1_17
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center