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Production and Physico-chemical Characterization of a Biosurfactant Produced by Pseudomonas aeruginosa OBP1 Isolated from Petroleum Sludge

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Abstract

Pseudomonas aeruginosa strain OBP1, isolated from petroleum sludge, was used to produce biosurfactant from a modified mineral salt medium with 2% n-hexadecane as sole source of carbon. The crude biosurfactant was fractionated using TLC and HPLC. Using FTIR spectroscopy, 1H NMR, and LC-MS analyses, the chemical structure of the purified fraction of crude biosurfactant was identified as rhamnolipid species. The LC-MS spectra show that monorhamnolipid (l-rhamnopyranosyl-β-hydroxydecanoyl-β- hydroxydecanoate, Rha-C10-C10) was produced in abundance with the predominant congener [M–H]− ions for l-rhamnopyranosyl-l-rhamnopyranosyl-3-hydroxydecanoyl-3-hydroxydecanoate (Rha-Rha-C10-C10). Seven different carbon substrates and five nitrogen sources were examined for their effect on rhamnolipid production. Using n-hexadecane (20 g/l) as carbon substrate and urea along with (NH4)2SO4 (2 g/l each) as nitrogen source was found to be the best, with a maximum yield of 4.8 g/l. The biosurfactant reduced the surface tension of water to 31.1 mNm−1 with a critical micelle concentration of 45 mg/l. The biosurfactant showed a better emulsifying activity against a variety of hydrocarbon and achieved a maximum emulsion index of 82% for diesel. The purified biosurfactant showed a significant antibacterial activity against Staphylococcus aureus at a minimum inhibitory concentration of 8 μg/ml.

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Acknowledgement

This work was financed by the Oil and Natural Gas Corporation, India. The authors would like to thank Dr. A. J. Thakur and Mr. S. Das for their technical support.

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  1. Department of Molecular Biology & Biotechnology, Tezpur University, Napaam, Tezpur, Assam, 784028, India

    Pranjal Bharali & Bolin Kumar Konwar

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  1. Pranjal Bharali
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Correspondence to Pranjal Bharali.

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Bharali, P., Konwar, B.K. Production and Physico-chemical Characterization of a Biosurfactant Produced by Pseudomonas aeruginosa OBP1 Isolated from Petroleum Sludge. Appl Biochem Biotechnol 164, 1444–1460 (2011). https://doi.org/10.1007/s12010-011-9225-z

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