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Media type:
E-Article
Title:
Development of a Biosensor Platform for Phenolic Compounds Using a Transition Ligand Strategy
Contributor:
Flachbart, Lion Konstantin
[Author];
Gertzen, Christoph Gerhard Wilhelm
[Author];
Gohlke, Holger
[Author];
Marienhagen, Jan
[Author]
imprint:
ACS, 2021
Published in:ACS synthetic biology 10(8), 2002–2014 (2021). doi:10.1021/acssynbio.1c00165
Language:
English
DOI:
https://doi.org/10.1021/acssynbio.1c00165
ISSN:
2161-5063
Origination:
Footnote:
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Description:
The time-consuming and laborious characterization of protein or microbial strain designs limits the development of high-performance biocatalysts for biotechnological applications. Here, transcriptional biosensors emerged as valuable tools as they allow for rapid characterization of several thousand variants within a very short time. However, for many molecules of interest, no specific transcriptional regulator determining a biosensor’s specificity is available. We present an approach for rapidly engineering biosensor specificities using a semirational transition ligand approach combined with fluorescence-activated cell sorting. In this two-step approach, a biosensor is first evolved toward a more relaxed-ligand specificity before using the resulting variant as the starting point in a second round of directed evolution toward high specificity for several chemically different ligands. By following this strategy, highly specific biosensors for 4-hydroxybenzoic acid, p-coumaric acid, 5-bromoferulic acid, and 6-methyl salicylic acid were developed, starting from a biosensor for the intracellular detection of trans-cinnamic acid.