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Nitro-aromatics are listed in carcinogenic, teratogenic, and mutagenic compounds list. p- nitro-aniline is one of them used as a precursor of various chemical compounds in many industries like dyes, drugs, paints and several others. These are mostly given out as an effluent in rivers, lakes or open passage of land which exert several hazards to living creatures and environment. Some of the organic compounds are stable in alkaline condition and persist longer in environment. Very few reports are elaborating bio-remediation in alkaline condition using different hydrocarbons. This study was planned to elaborate mechanism of detoxification and searching the potential of decontamination of p-nitro-aniline in alkaline condition by experimental microbial strain. The bacterial strain pseudomonas DL17 was isolated from alkaline Lake Lonar, Buldana, (MS.) India; and employed in this experiment considering its indigenous property to tolerate the alkaline pH. It also showed resistance to p-nitro-aniline with its raising concentrations on testing after adaptation. The experimental microbial stain showed 100% biodegradation of (500 mg/L) p-nitro-aniline within 48h. On shaking incubator with 110 rpm and at 32 °C optimum temperature. The centrifugate obtained after spinning at 10,000 g by cold centrifuge was used for solvent extraction. Generally, ethyl acetate or DCM was used for solvent extraction. The estimation of residual remains of p-nitro aniline by 6h. intervals was carried after removal of flasks from shaking incubator and centrifugation. At the optimum temperature and pH experiments were carried after knowing the resistance to experimental contaminant range (100-400 mg/L) of p-nitro aniline one month and further extended to 500 mg/L for 15days more. The residual metabolites were purified by column chromatography and various spectrometric studies such as UV-Vis spectroscopy, HNMR, FTIR and GCMS revealed that p-Phenylenediamine, acetanilide, aniline, acetaminophen, catechol, p-bezoquinone, cis-cis muconate as a metabolites. On the basis of the metabolites isolated and characterized by different spectroscopic studies the bio-catalytic mechanism was deduced. The induced enzymes such as nitroreductase, catalase, peroxidase, acetanilide hydroxylase, super oxide dismutase, catechol 1, 2 dioxygenase, catechol 2, 3 dioxygenase has commercial importance in biochemical industries. Induction of such biotransformation enzymes and consumption of p-nitro aniline concentration in experiments makes sure that this microbial strain pseudomonas DL17 can be employed for decontamination of nitro aniline polluted sites as well as isolation of such metabolites characterized and enzymes studied.

Biotransformation of nitro aromatic amines in artificial alkaline habitat by pseudomonas DL17