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000894465 1001_ $$0P:(DE-Juel1)162145$$aFlachbart, Lion Konstantin$$b0
000894465 245__ $$aDevelopment of a Biosensor Platform for Phenolic Compounds Using a Transition Ligand Strategy
000894465 260__ $$aWashington, DC$$bACS$$c2021
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000894465 520__ $$aThe 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.
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000894465 7001_ $$0P:(DE-Juel1)174133$$aGertzen, Christoph Gerhard Wilhelm$$b1
000894465 7001_ $$0P:(DE-Juel1)172663$$aGohlke, Holger$$b2
000894465 7001_ $$0P:(DE-Juel1)144031$$aMarienhagen, Jan$$b3$$eCorresponding author
000894465 773__ $$0PERI:(DE-600)2644383-1$$a10.1021/acssynbio.1c00165$$gp. acssynbio.1c00165$$n8$$p2002–2014$$tACS synthetic biology$$v10$$x2161-5063$$y2021
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