Development of engineered sensor perceiving gaseous toluene signal in the cyanobacterium Synechocystis sp. PCC 6803

Autor: Ryo Morioka, Yoshihiro Shiraiwa, Muhammad Junaid, Yu Inaba, Iwane Suzuki
Rok vydání: 2017
Předmět:
Zdroj: Journal of Applied Phycology. 30:71-78
ISSN: 1573-5176
0921-8971
Popis: Microalgae have been used to produce useful compounds. In many cases in which target compounds are produced from CO2 via photosynthesis, there is a trade-off between the proliferation of cells and the production of the target compounds. This is due to carbon partitioning between the synthesis of cellular components and the target compounds, and/or the toxicity of the target products. Therefore, there is a requirement to develop a gene switch that can modulate the metabolism from the growth phase of cells to the production phase of the target molecules. In this study, we attempted to develop an artificial sensory protein that can respond to insoluble gaseous compounds and regulate gene expression in the model cyanobacterium Synechocystis sp. PCC 6803. We constructed a chimeric histidine kinase by fusing the signal input domain of TodS, a toluene sensor from Pseudomonas putida, and the kinase domain of SphS, a phosphate-deficient sensor from Synechocystis. Because TodS contains two kinase domains, we substituted either of the kinase domains of TodS with the kinase domain of SphS to obtain TodSS_SphS and TodSL_SphS. We introduced a gene for the chimeric sensor to Synechocystis cells and evaluated their response to toluene by measuring alkaline phosphatase (AP) activity and phoA mRNA expression. Although the cells harboring TodSL_SphS did not exhibit AP activity, both in the presence or absence of toluene, the cells harboring TodSS_SphS accumulated the transcript of the phoA gene after 5-h exposure to toluene, and only exhibited AP activity after exposure to toluene.
Databáze: OpenAIRE
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