Applied Biochemistry and Biotechnology, Vol.175, No.4, 1789-1804, 2015
Biodegradation of Insecticide Monocrotophos by Bacillus subtilis KPA-1, Isolated from Agriculture Soils
Twenty bacterial strains, which are capable of degrading monocrotophos, were isolated from five soil samples collected from agriculture soils in India. The ability of the strains to mineralize monocrotophos was investigated under different culture conditions. A potential strain degrading monocrotophos was selected and named KPA-1. The strain was identified as a Bacillus subtilis on the basis of the results of its cellular morphology, physiological and chemotaxonomic characteristics, and phylogenetic conclusion of 16S ribosomal DNA (rDNA) gene sequences. Organophosphate hydrolase (opdA gene) involved in the initial biodegradation of monocrotophos in KPA-1 was quantitatively expressed, which was a constitutively expressed cytosolic enzyme. RT-qPCR data revealed that KPA-1 harboring opdA gene in an early stage was significantly downregulated from opdA gene in a degradation stage (1.5 fold more) with a p value of 0.0375 (p<0.05). We have optimized culture conditions for the efficient degradation (94.2 %) of monocrotophos under aerobic conditions. Growth and degradation kinetic studies proved that KPA-1 was able to grow in minimal salt medium containing 1000 ppm monocrotophos as the only carbon source. Hence, KPA-1 culture has a great potential utility for the bioremediation of agriculture soils contaminated with organophosphorus pesticides, particularly monocrotophos.
Keywords:Bacillus subtilis;Biodegradation;Monocrotophos;Organophosphate hydrolase;Organophosphorus pesticide;Soil contamination