Abstract
The current investigation is taken up with the aim of studying repeated batch and continuous degradation of Endosulfan, using Ca-alginate immobilized cells of Pseudomonas aeruginosa isolated from an agricultural soil. The work involves the study of genes and enzymes involved in the degradation of the pesticide and was carried out with an objective of reducing the toxicity of Endosulfan by degrading it to less toxic metabolites. The long-term stability of Endosulfan degradation was studied during its repeated batch degradation, carried out over a period of 35 days. Immobilized cells of Ps. aeruginosa were able to show 60 % degradation of Endosulfan at the end of the 35th cycle with a cell leakage of 642 × 10(4) Cfu/mL. During continuous treatment, with 2 % concentration of Endosulfan, 100 % degradation was recorded up to 100 mL/h flow rate and with 10 % concentration of the Endosulfan, and 100 and 85 % degradation was recorded at 20 mL/h flow rate and 100 mL/h flow rate, respectively. After degradation of Endosulfan, products were extracted from a large amount of spent medium using two volumes of ethyl acetate and subjected to the LC-MS analysis. Endosulfan lactone and Endosulfan ether were the products of degradation detected by the LCMS analysis. Plasmid curing experiments indicated that genes responsible for the degradation of Endosulfan are present on the chromosome and not on the plasmid, as growth of Ps. aeruginosa was observed on modified non-sulfur medium with Endosulfan after the plasmid was cured with ethidium bromide. The results of PCR indicated that there is no amplified product of ~1350 bp expected for esd gene, in Ps. aeruginosa, although there were some non-specific bands. Enzymatic degradation studies indicated that the enzymes involved in the degradation of Endosulfan are intracellular. With this investigation, it was indicated that immobilized cells of Ps. aeruginosa have the potential to be used in the bioremediation of water contaminated with Endosulfan.