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Molecular Characterization of Bacteria Capable of Organophosphate Degradation

A Ratna Kumari1, K Sobha2, K Mounika1, G J Nageswara Rao1, and M Ashok1
1. Department of Biotechnology, Bapatla Engineering College, Bapatla-522 101, AP, India
2. Department of Biotechnology, RVR & JC College ofEngineering,Chowdavaram, Guntur-522 019, AP, India
Abstract—Synthetic organophosphorus compounds (OP) are widely used in agriculture to control agricultural and household pests. The excessive use of OP compounds have generated a number of environmental problems such as contamination of air, water and terrestrial ecosystems, harmful effects on different biota, and disruption of biogeochemical cycling. Therefore, there is a need for economical and dependable methods of OPs detoxification from the environment. Conventional methods (chemical treatment, incineration and landfills) are having a number of disadvantages when compared to biological methods. Soil microflora is one of the basic eco-friendly agents for detoxification of pesticides. OP pesticide-degrading bacteria have been isolated by using soil enrichment process with organophosphates. From pesticide enriched soil through serial dilution pure isolated colonies were obtained. For molecular identification of the isolated strains genomic DNA was isolated and analyzed by agarose gel electrophoresis. The 16S rRNA gene was amplified through PCR using forward primer 5’ AGAGTTTGAT CCTGGCTCAG 3’ and reverse primer 5’ GACGGGCRGTGWGTR CA 3’. The amplified product was purified & the gene was sequenced using sangersdideoxy method. The 1330 nucleotide sequence of the gene was compared with known nucleotide sequence using BLASTn and this gene showed 99% similarity with the 16S rRNA gene of the Pseudomonas putida. Finally, the phylogenetic analysis was done.To analyze the degradation capability, the phosphate levels are estimated by Fiske-Subbarow method. The phosphate levels are increased due to detoxification of organophosphorus compounds. 

Index Terms—organophosphate (OP), pseudomonas putida, BLASTn, phylogenetic analysis

Cite: A Ratna Kumari, K Sobha, K Mounika, G J Nageswara Rao, and M Ashok, "Molecular Characterization of Bacteria Capable of Organophosphate Degradation," International Journal of Life Sciences Biotechnology and Pharma Research, Vol. 1, No. 3, pp. 64-72, July 2012.
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