A unified design allows fine-tuning of biosensor parameters and application across bacterial species

Sonntag, Christiane Katharina; Marienhagen, Jan; Maass, Celine; Flachbart, Lion Konstantin; Vogt, Michael

doi:10.1016/j.mec.2020.e00150

Journal Article

FZJ-2020-04120

A unified design allows fine-tuning of biosensor parameters and application across bacterial species

Sonntag, C. K.FZJ* ; Flachbart, L. K. ; Maass, C. ; Vogt, M. ; Marienhagen, J. (Corresponding author)FZJ*

2020
Elsevier Amsterdam [u.a.]

Metabolic engineering communications 11, e00150 - (2020) [10.1016/j.mec.2020.e00150]

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Please use a persistent id in citations: http://hdl.handle.net/2128/25989 doi:10.1016/j.mec.2020.e00150

Abstract: In recent years, transcriptional biosensors have become valuable tools in metabolic engineering as they allow semiquantitative determination of metabolites in single cells. Although being perfectly suitable tools for high-throughput screenings, application of transcriptional biosensors is often limited by the intrinsic characteristics of the individual sensor components and their interplay. In addition, biosensors often fail to work properly in heterologous host systems due to signal saturation at low intracellular metabolite concentrations, which typically limits their use in high-level producer strains at advanced engineering stages.We here introduce a biosensor design, which allows fine-tuning of important sensor parameters and restores the sensor response in a heterologous expression host. As a key feature of our design, the regulator activity is controlled through the expression level of the respective gene by different (synthetic) constitutive promoters selected for the used expression host. In this context, we constructed biosensors responding to basic amino acids or ring-hydroxylated phenylpropanoids for applications in Corynebacterium glutamicum and Escherichia coli. Detailed characterization of these biosensors in liquid cultures and during single-cell analysis using flow cytometry showed that the presented sensor design enables customization of important biosensor parameters as well as application of these sensors in relevant heterologous hosts.

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