The Journal of Biological Physics and Chemistry


Volume 6, Number 3, p.p. 119-123

A cellular bioassay for TNT detection using engineered Pseudomonas sp. strain TM101 for systematic bioremediation

Hitoshi Asakawa, Toshinari Maeda, Hiroaki I. Ogawa and Tetsuya Haruyama

Department of Biological Functions and Engineering, Kyushu Institute of Technology, Kitakyushu Science and Research Park, Kitakyushu, Fukuoka 808-0196, Japan

We have previously developed a Pseudomonas sp.-based bioremediation process for 2,4,6-trinitrotoluene (TNT)-polluted soil. However, an easy assay is required in order to systemize the bioremediation. Hence to complement the microorganism-based bioremediation process, an engineered microorganism-based bioassay for (TNT) detection was developed. We have successfully bred transposon mutant TM101 from the wild-type Pseudomonas sp. strain TM15 for this purpose. The mutant strain TM101 can degrade TNT and accumulate nitrite through the metabolic pathway of TNT transformation. In our present assay, the biologically accumulated nitrite is employed as a marker, and is measured to determine the concentration of TNT contaminant. The results indicate that the present cellular bioassay has sufficiently high TNT selectivity and is easy to perform. The bioassay can be applied routinely in TNT field surveys. Our previously developed bioremediation method and the present cellular bioassay will demonstrate their full effect when they are employed as one process for the decontamination of polluted ground.

Keywords: bioremediation, cell-based bioassay, cellular bioassay, engineered cell, nitrite accumulation, 2,4,6-trinitrotoluene

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