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Engineering chimeric two-component system into Escherichia coli from Paracoccus denitrificans to sense methanol

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Abstract

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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Authors and Affiliations

  1. Department of Chemical Engineering, University of Ulsan, Ulsan, 680-749, Korea

    Vidhya Selvamani, Irisappan Ganesh, Murali kannan Maruthamuthu & Soon Ho Hong

  2. Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), Ulsan, 681-802, Korea

    Gyeong Tae Eom

  3. Department of Green Chemistry and Environmental Biotechnology, Korea University of Science and Technology (UST), Daejeon, 305-350, Korea

    Gyeong Tae Eom

Authors
  1. Vidhya Selvamani
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  2. Irisappan Ganesh
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  3. Murali kannan Maruthamuthu
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  4. Gyeong Tae Eom
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  5. Soon Ho Hong
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Correspondence to Soon Ho Hong.

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Selvamani, V., Ganesh, I., Maruthamuthu, M.k. et al. Engineering chimeric two-component system into Escherichia coli from Paracoccus denitrificans to sense methanol. Biotechnol Bioproc E 22, 225–230 (2017). https://doi.org/10.1007/s12257-016-0484-y

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  • DOI: https://doi.org/10.1007/s12257-016-0484-y

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