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bioRxiv · 10.1101/2025.04.03.647055

Autoinduction expression systems by AHL-mediated quorum-sensing circuits in Synechococcus elongatus PCC 7942

Kokarakis, E.·Santos-Merino, M.·Ghaffarinasab, S.·Vocelle, D.·Ducat, D.
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Abstract

Despite significant potential for cyanobacteria as sustainable bioproduction chases, there are limited examples of scaled cyanobacterial bioproduction. In part, this is because most cyanobacterial species are poorly adapted to bioreactor cultivation conditions and lack features that facilitate biomass growth and harvesting at scale. We explored quorum sensing (QS) pathways derived from heterotrophic microbes as a method for autoinduction of gene expression circuits coordinated to population density in cyanobacteria. Here, we integrated genetic modules designed to produce and detect the diffusible QS signal, acyl-homoserine lactones (AHLs), in the cyanobacterial model, Synechococcus elongatus PCC 7942 (S. elongatus). We demonstrate that S. elongatus heterologously produces sufficient AHL signals to activate gene expression in a dose-dependent and population density-responsive manner. A hybrid combination of AHL synthesis enzyme from Vibrio fischeri (Lux system) with the transcription factor receiver from Pseudomonas aeruginosa (Las system) provides an ideal activation ratio and mitigates toxicity observed with some AHL systems. As a proof of concept, we coupled the QS pathway to the expression of a cell division inhibitory gene, cdv3, facilitating late-phase cell elongation, cell sedimentation, and improved biomass recovery. Our findings provide a foundation for the development of auto-induction systems leverageable to improve cyanobacterial biotechnology applications.

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Kokarakis, E., Santos-Merino, M., Ghaffarinasab, S., Vocelle, D., Ducat, D.. 2025-04-03. Autoinduction expression systems by AHL-mediated quorum-sensing circuits in Synechococcus elongatus PCC 7942. https://doi.org/10.1101/2025.04.03.647055

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