Engineering Chimeric Two-component System into Escherichia coli from Paracoccus denitrificans to Sense Methanol
Vidhya Selvamani, University of Ulsan, Ulsan, Republic of Korea | Irisappan Ganesh, University of Ulsan, Ulsan, Republic of Korea | Murali kannan Maruthamuthu, University of Ulsan, Ulsan, Republic of Korea | Eom, G.T., Korea Research Institute of Chemical Technology (KRICT), Ulsan, Republic of Korea | Hong, S.H., University of Ulsan, Ulsan, Republic of Korea
Escherichia coli does not have the methanol sensing apparatus, was engineered to sense methanol by employing chimeric two-component system (TCS) strategy. A chimeric FlhS/EnvZ (FlhSZ) chimeric histidine kinase (HK) was constructed by fusing the sensing domain of Paracoccus denitrificans FlhS with the catalytic domain of E. coli EnvZ. The constructed chimeric TCS FlhSZ/OmpR could sense methanol by the expression of ompC and gfp gene regulated by ompC promoter. Real-time quantitative PCR analysis and GFP-based fluorescence analysis showed the dynamic response of the chimeric TCS to methanol. The expression of ompC and the gfp fluorescence was maximum at 0.01 and 0.5% of methanol, respectively. These results suggested that E. coli was successfully engineered to sense methanol by the introduction of chimeric HK FlhSZ. This strategy can be employed for the construction of several chimeric TCS based bacterial biosensors for the development of biochemical producing recombinant microorganisms.
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