Isolation of a chlorpyrifos-degrading bacterium, Sphingomonas sp. strain Dsp-2, and cloning of the mpd gene

Abstract

A highly effective chlorpyrifos-degrading bacterium strain Dsp-2 was isolated from the polluted treatment system of a chlorpyrifos manufacturer. This strain was preliminarily identified as Sphingomonas sp. based on its morphological, physiological and biochemical tests as well as 16S rDNA analysis. It utilized chlorpyrifos as its sole source of carbon for growth, by hydrolyzing chlorpyrifos to 3,5,6-trichloro-2-pyridinol (TCP). It could also utilize parathion, parathion-methyl, fenitrothion and profenofos, but not phoxin and triazophos. Bioremediation of chlorpyrifos-contaminated soil was examined using Dsp-2. Dsp-2 addition to soil treated with 100mgkg(-1) chlorpyrifos resulted in a higher degradation rate than control soils without inoculation. The moderate pH, moisture and inoculum density could have promoted degradation. The gene encoding the chlorpyrifos hydrolytic enzyme was cloned by PCR. Although BLAST sequence search results indicated that this gene has 99% similarity to mpd (a gene encoding the parathion-methyl hydrolyzing enzyme in Plesiomonas sp. M6), its hydrolytic efficiency for chlorpyrifos was significantly greater than the wild-type mpd from strain M6.