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Pseudomonas sp. JHN decolourized and biotransformed 4-chloro-2-nitrophenol (4C2NP) in the presence of additional carbon source. The effect of the various concentrations of the 4C2NP was studied on the decolourization of 4C2NP by Pseudomonas sp. JHN. It was observed that strain JHN decolourized and biotransformed 4C2NP up to concentration of 0.6 mM. Gas chromatography and gas chromatography-mass spectrometry detected 5-chloro-2-methylbenzoxazole as a major metabolite of the co-metabolism of 4C2NP. Furthermore, strain JHN exhibits positive chemotaxis toward 4C2NP based on the drop plate and capillary assays. This is the first report of the chemotaxis toward 4C2NP by any bacterium.