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@ARTICLE{Flachbart:894465,
author = {Flachbart, Lion Konstantin and Gertzen, Christoph Gerhard
Wilhelm and Gohlke, Holger and Marienhagen, Jan},
title = {{D}evelopment of a {B}iosensor {P}latform for {P}henolic
{C}ompounds {U}sing a {T}ransition {L}igand {S}trategy},
journal = {ACS synthetic biology},
volume = {10},
number = {8},
issn = {2161-5063},
address = {Washington, DC},
publisher = {ACS},
reportid = {FZJ-2021-03241},
pages = {2002–2014},
year = {2021},
note = {Biotechnolgie},
abstract = {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.},
cin = {IBG-1},
ddc = {570},
cid = {I:(DE-Juel1)IBG-1-20101118},
pnm = {2171 - Biological and environmental resources for
sustainable use (POF4-217) / DFG project 417919780 - Zentrum
für strukturelle Studien},
pid = {G:(DE-HGF)POF4-2171 / G:(GEPRIS)417919780},
typ = {PUB:(DE-HGF)16},
pubmed = {34369151},
UT = {WOS:000687920000019},
doi = {10.1021/acssynbio.1c00165},
url = {https://juser.fz-juelich.de/record/894465},
}