Abstract
Due to its recalcitrant and carcinogenic nature, the presence of methyl orange (MO) in the environment is a serious threat to human and animal life and is also toxic to plants. MO being recalcitrant cannot be effectively reclaimed from industrial effluents through physical and chemical approaches. Biological methods on the other hand have the potential to degrade such dyes because of their compatibility with nature and low chances of adverse effects on the environment. Bacteria, due to their fast growth rate and capability of surviving in extreme environments can effectively be used for this purpose. In the current research study, Pseudomonas aeruginosa was isolated and characterized using 16rRNA from textile wastewater. In the preliminary tests it was found that Pseudomonas aeruginosa has the ability to degrade and mineralize methyl orange effectively. The physicochemical conditions were then optimized, in order to get maximum degradation of MO which was achieved at 37 °C, a pH of 7, a low salt concentration of 0.1 g/15 mL, a high carbon source of 0.6 g/15 mL, and 72 h experimental time. In a single set of experiments where all these optimum conditions were combined, 88.23% decolorization of the selected dye was achieved. At the end of the experimental cycle, the aliquots were homogenized and filtered. The filtrates were subjected to FTIR and GC-MS analysis where azo linkage breaking was confirmed from the FTIR spectra. The filtrates were then extracted with ethyl acetate and then passed through a silica gel column. On the basis of Rf value (TLC plates used) similar fraction were combined which were then subjected to NMR analysis. The compounds detected through GC-MS, peaks were not observed in proton and C-13 NMR. Instead, solvent and some impurity peaks were present, showing that complete mineralization of the dye had occurred due to the action of different bacterial enzymes such as azoreductase, peroxidases, and classes on MO. The prosed mechanism of complete mineralization is based on spectral data that needs to be verified by trapping the individual step products through the use of appropriate inhibitors of individual enzymes.