Development of a Biosensor Platform for Phenolic Compounds Using a Transition Ligand Strategy
| Autor: | Flachbart, Lion Konstantin, Gertzen, Christoph Gerhard Wilhelm, Gohlke, Holger, Marienhagen, Jan |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: | |
| Zdroj: | ACS synthetic biology 10(8), 2002-2014 (2021). doi:10.1021/acssynbio.1c00165 ACS synthetic biology 10(8), 2002–2014 (2021). doi:10.1021/acssynbio.1c00165 |
| DOI: | 10.1021/acssynbio.1c00165 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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