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Chimeric LysR-type transcriptional biosensors for customising ligand specificity profiles towards flavonoids.

Research paper by Brecht B De Paepe, Jo J Maertens, Bartel B Vanholme, Marjan M De Mey

Indexed on: 20 Dec '18Published on: 20 Dec '18Published in: ACS Synthetic Biology



Abstract

Transcriptional biosensors enable key applications in both metabolic engineering and synthetic biology. Due to nature's immense variety of metabolites, these applications require biosensors with a ligand specificity profiles customised to the researcher's needs. In this work, chimeric biosensors were created by introducing parts of a donor regulatory circuit from Sinorhizobium meliloti, delivering the desired luteolin-specific response, into a non-specific biosensor chassis from Herbaspirillum seropedicae. Two strategies were evaluated for the development of chimeric LysR-type biosensors with customised ligand specificity profiles towards three closely-related flavonoids, naringenin, apigenin and luteolin. In the first strategy, chimeric promoter regions were constructed at the biosensor effector module, while in the second strategy, chimeric transcription factors were created at the biosensor detector module. Via both strategies, the biosensor repertoire was expanded with luteolin-specific chimeric biosensors demonstrating a variety of response curves and ligand specificity profiles. Starting from the non-specific biosensor chassis, a shift from 27.5% to 95.3% luteolin specificity was achieved with the created chimeric biosensors. Both strategies provide a compelling, faster and more accessible route for the customisation of biosensor ligand specificity, compared to de novo design and construction of each biosensor circuit for every desired ligand specificity.