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Degradation of Phenol by Mixed Culture of Locally Isolated Pseudomonas Species

Sadia Afrin Jame
Department of Environmental Sciences, Jahangirnagar University, Savar, Dhaka 1342, Bangladesh

A.K.M. Rashidul Alam
Department of Environmental Sciences, Jahangirnagar University, Savar, Dhaka 1342, Bangladesh

A.N.M. Fakhruddin
Department of Environmental Sciences, Jahangirnagar University, Savar, Dhaka 1342, Bangladesh

Md. Khorshed Alam
Institute of Food & Radiation Biology (IFRB), Atomic Energy Research Establishment (AERE), Ganakbari, Savar, Dhaka 1344, Bangladesh

Abstract/ Executive Summary:

Textile, pharmaceuticals and automobile waste most often contain phenolic wastes. An attempt was made to degrade phenol using locally isolated bacteria. Moreover, as in nature pure culture is rarely found rather than mixed culture, therefore, attempt was also made to improve the degradation using mixed culture of Pseudomonas species. All isolates could completely degrade phenol up to 600 ppm. Isolate Pseudomonas FA degraded 800 ppm phenol completely in 72 hours, but the isolates Pseudomonas SA, TK and KA degraded only 39.33, 43.83 and 33.16% of 800 ppm phenol respectively in 96 hours. Complete removal time was also shorter for the isolate Pseudomonas FA compares to the other isolates. Patterns of growth were similar for all of the isolates, but maximum growth was found with the isolate FA on 600 ppm phenol. Complete degradation time was decreased with mixed culture and removal rate of phenol was 25 ppm/h in mixed culture of all combinations and was higher than that of the single culture of the isolates. In mixed culture study, the growth of bacteria was also increased.

Keywords: Biodegradation; Phenol; Mixed culture; Pseudomonas

Location: Food Microbiology and Industrial Irradiation Division, Institute of Food and Radiation Biology, Bangladesh Atomic Energy Research Establishment, Savar

Start Date:

End Date:

Program Area: Risk Management in Agriculture

Commodity: Bio fertilizer

Objectives:

The study aims to study the rate of degradation of phenol when supplied as a sole source of carbon at various concentrations by the bacterial isolates and to evaluate the degradation capability of bacterial isolates by mixed culture in order to determine their practical applicability.

Methodology:

Microorganisms: Pseudomonas putida CP1 and Pseudomonas putida A (a) was obtained from the stock culture of Food Microbiology and Industrial Irradiation Division, Institute of Food and Radiation Biology, Bangladesh Atomic Energy Research Establishment, Savar, Dhaka, Identification of these two isolates were described elsewhere. The remaining four locally isolated bacterial were Pseudomonas SA, TK, KA and FA. Identification of the local isolates was also described. Chemicals and media: Phenol used in this study was obtained from Sigma-Aldrich Co., UK. Bacteria capable of degrading phenol were cultured on minimal medium comprised K₂HPO₄- 4.36 g/l; Na₂HPO₄-3.45 g/l; NH₄Cl- 1.0 g/l; MgSO₄.6H₂O – 0.912 g/l; trace salt solution- 1ml/l. The pH of the medium was adjusted to 7.0 with 2M NaOH. The trace salt solution contained 4.77 g-CaCl₂. 2H₂O; 0.37 g -FeSO₄. 7H₂O; 0.37 g-CoCl₂.6H₂O, 0.10 g-MnCl₂.4H₂O and 0.02 g-NaMoO₄. 2H₂O in 100 ml water. Inoculums: Cells from the Pseudomonas minimal medium were used to inoculate nutrient broth (15 ml). The harvested cells were centrifuged at 5000 rpm for 10 minutes and washed twice with 0.01M sodium phosphate buffer and final pellet resuspended in the same buffer. Bacterial suspension (10⁷/ml) was used to inoculate 95 ml sterile minimal medium containing phenol in 250 ml conical flasks to give the appropriate final concentration of the added substrates. After inoculation, flasks were incubated in an orbital shaker at 120 rpm 30°C. Samples were aseptically removed at regular intervals and analyzed for growth, substrate removal and pH. Growth of the organisms was monitored by measuring optical density at 660 nm. For mixed culture study, equal amounts of different inoculums were taken and the ultimate volume was fixed to 5 ml in 250 ml conical flasks to give the appropriate final concentration of the added inoculums. After inoculation, flasks were incubated in an orbital shaker at 120 rpm at 30°C. Rest of all the procedures were followed as above. Chemical analyses: Samples were then centrifuged at 5000 rpm for 10 minutes; the supernatants were then analyzed for phenol. Phenol concentration was determined by using the 4-aminoantipyrene colorimetric method.

Source of Information: J Bioremed Biodegrad 1:102

Article Link (Online Journal): doi:10.4172/2155-6199.1000102

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Pseudomonas sp. SA, Pseudomonas sp. TK and Pseudomonas sp. KA were able to degrade 600 ppm and Pseudomonas sp. FA was able to degrade phenol up to 800 ppm of phenol. In mixed culture study, the growth of bacteria was increased and degradation rate was also increased.

Knowledge Management: Journal