CHARACTERIZATION OF 2,4,6-TRINITROTOLUENE (TNT)-METABOLIZING STRAIN PSEUDOMONAS AERUGINOSA SP TUHP1 ISOLATED FROM TNT-POLLUTED SOILS IN THEVELLORE DISTRICT, TAMILNADU,INDIA
Keywords:Bioremediation, Biodegradation, Biotransformation, Sequencing
Objective: The main objective was to evaluate the degradative properties of Pseudomonas aeruginosa sp TUHP1 isolated from TNT-Polluted soils in the Vellore District, Tamil Nadu, India.
Methods: Among the 3 bacterial genera isolated from different soil samples, one potent TNT degrading strain Pseudomonas aeruginosa sp TUHP1 was identified. The morphological, physiological and the biochemical properties of the strain Pseudomonas aeruginosa sp TUHP1 was confirmed by conventional methods and genotypic characterization was carried out using 16S r-DNA partial gene amplification and sequencing. The broken down by products of DNT in the extract was determined by Gas Chromatogram- Mass spectrometry (GC-MS). Supernatant samples from the broth studied at 24 h interval were analyzed by HPLC analysis and the effect on various nutritional and environmental factors were analysed and optimized for the isolate.
Results: Out of three isolates one strain TUHP1 were found to have potent efficiency to degrade TNT and revealed the genus Pseudomonas. 16S rDNA gene sequence analysis showed highest homology (99%) with Pseudomonas aeruginosa DS10 and was assigned as Pseudomonas aeruginosa sp TUHP1. Based on the energy of the predicted models, the secondary structure predicted by MFE showed the more stable structure with a minimum energy. Products of TNT Transformation showed colour change in the medium during cultivation. TNT derivates such as 2HADNT and 4HADNT were detected by HPLC chromatogram and 2ADNT, 4ADNT by GC/MS analysis.
Conclusion: Hence this study presents the clear evidence for the biodegradation process of TNT by strain Pseudomonas aeruginosa sp TUHP1.
Keywords: Bioremediation. Biodegradation. Biotransformation. Sequencing.
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