%0 Journal Article
%A Flachbart, Lion Konstantin
%A Gertzen, Christoph Gerhard Wilhelm
%A Gohlke, Holger
%A Marienhagen, Jan
%T Development of a Biosensor Platform for Phenolic Compounds Using a Transition Ligand Strategy
%J ACS synthetic biology
%V 10
%N 8
%@ 2161-5063
%C Washington, DC
%I ACS
%M FZJ-2021-03241
%P 2002–2014
%D 2021
%Z Biotechnolgie
%X 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.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ 34369151
%U <Go to ISI:>//WOS:000687920000019
%R 10.1021/acssynbio.1c00165
%U https://juser.fz-juelich.de/record/894465