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Construction of 2,4,6-Trinitrotoluene Biosensors with Novel Sensing Elements from Escherichia coli K-12 MG1655

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Abstract

Detection of 2,4,6-trinitrotoluene (TNT) has been extensively studied since it is a common explosive filling for landmines, posing significant threats to the environment and human safety. The rapid advances in synthetic biology give new hope to detect such toxic and hazardous compounds in a more sensitive and safe way. Biosensor construction anticipates finding sensing elements able to detect TNT. As TNT can induce some physiological responses in E. coli, it may be useful to define the sensing elements from E. coli to detect TNT. An E. coli MG1655 genomic promoter library containing nearly 5,400 elements was constructed. Five elements, yadG, yqgC, aspC, recE, and topA, displayed high sensing specificity to TNT and its indicator compounds 1,3-DNB and 2,4-DNT. Based on this, a whole cell biosensor was constructed using E. coli, in which green fluorescent protein was positioned downstream of the five sensing elements via genetic fusion. The threshold value, detection time, EC200 value, and other aspects of five sensing elements were determined and the minimum responding concentration to TNT was 4.75 mg/L. According to the synthetic biology, the five sensing elements enriched the reservoir of TNT-sensing elements, and provided a more applicable toolkit to be applied in genetic routes and live systems of biosensors in future.

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Acknowledgments

We would like to thank Professor Wang Hengliang (Beijing Institute of Bioengineering) for help and guidance in this study. This study was funded by the synthetic biology project of Chinese National High Technology Research and Development Program (863 Program). Project Number 2012AA02A706.

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Authors and Affiliations

  1. Beijing Institute of Biotechnology, Beijing, China

    Junjie Tan, Jingyi Ling, Guolong Qu, Gang Liu & Huipeng Chen

  2. College of Life Sciences, Anhui University, Hefei, China

    Naipeng Kan & Yu Shao

  3. College of Life Sciences, Jilin University, Changchun, China

    Wei Wang

  4. ShenYang Pharmaceutical University, Shenyang, China

    Jing Jin

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  1. Junjie Tan
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Correspondence to Gang Liu or Huipeng Chen.

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Tan, J., Kan, N., Wang, W. et al. Construction of 2,4,6-Trinitrotoluene Biosensors with Novel Sensing Elements from Escherichia coli K-12 MG1655. Cell Biochem Biophys 72, 417–428 (2015). https://doi.org/10.1007/s12013-014-0481-8

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  • DOI: https://doi.org/10.1007/s12013-014-0481-8

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