Development of Transcription Factor-Based Biosensors for Butanol Detection in Synechocystis PCC 6803

Sammanfattning

Butanol is an important biofuel candidate due to its favorable fuel properties, and microbial production in cyanobacteria provides a potential carbon-neutral alternative to fossil-based transportation fuels. To enable high throughput screening of engineered strains for improved butanol production, transcription-factor based biosensors can be used. In this study two biosensor system based on transcription-factors AlkS and BmoR were investigated for adaption in the cyanobacterium Synechocystis PCC 6803. The BmoR-based biosensor variants were constructed via Gibson assembly, yet these constructs could not be conclusively verified and were not further evaluated. An AlkS-based biosensor was successfully assembled, cloned in E. coli, and transferred to Synechocystis. The biosensor was tested in both species under varying butanol concentrations, but no apparent biosensor signal was observed. The results from this project suggest that further optimization and characterization of the biosensor systems are needed before they can be applied as screening tools in butanol-producing strains.