TY  - JOUR
AU  - Flachbart, Lion Konstantin
AU  - Gertzen, Christoph Gerhard Wilhelm
AU  - Gohlke, Holger
AU  - Marienhagen, Jan
TI  - Development of a Biosensor Platform for Phenolic Compounds Using a Transition Ligand Strategy
JO  - ACS synthetic biology
VL  - 10
IS  - 8
SN  - 2161-5063
CY  - Washington, DC
PB  - ACS
M1  - FZJ-2021-03241
SP  - 2002–2014
PY  - 2021
N1  - Biotechnolgie
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - 34369151
UR  - <Go to ISI:>//WOS:000687920000019
DO  - DOI:10.1021/acssynbio.1c00165
UR  - https://juser.fz-juelich.de/record/894465
ER  -