Polyaromatic hydrocarbons (PAHs) are uncharged, non-polar molecules generated from natural and anthropogenic activities, where the emissions from anthropogenic activities predominate. Chrysene is a high molecular weight PAH, which is found to be highly recalcitrant and mutagenic in nature. The aim of this study was to isolate chrysene-degrading microorganisms from oil-contaminated water and to enhance their degradative conditions using design expert. From the various samples collected, 19 bacterial strains were obtained through enrichment culture and the one which showed highest activity was identified by 16S rRNA sequencing as Bacillus halotolerans. Under optimum conditions of 100 mg/l chrysene concentration, 1,000 mg/l nitrogen source, pH 6, B. halotolerans exhibited 90% chrysene degradation on sixth day. Positive results for the enzymes laccase and catechol 1,2 dioxygenase confirmed the ability for chrysene degradation by the isolated strain. Major metabolic intermediate determined in GCMS analysis was diisooctyl phthalate. Hence it can be concluded that Bacillus halotolerans can be a promising candidate for the removal of HMW hydrocarbons from contaminated environments.
Biosurfactants were found to be a suitable candidate for environment clean-up and as an alternative to conventional treatment systems. In the present study, a polyaromatic hydrocarbon degrading organism Bacillus halotolerans was screened for its ability to produce biosurfactant during hydrocarbon degradation. The bio-surfactant activity of the organism was screened by using various methods like oil spreading technique, emulsification assay, foam height analysis and parafilm-M test. The design expert software was used to optimize the suitable conditions for the production of biosurfactants. The optimum conditions were determined as pH 6, Chrysene-100 ppm, nitrogen source -1000 ppm and at 144 hrs using the design of experiments. The organism showed good oil degradation capacity and maximum growth was achieved in 6 days. Biosurfactant obtained from the biodegradation medium was confirmed to be lipopeptide using TLC, FTIR and GCMS.
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