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Bacterial consortium expressing surface displayed, intra- and extracellular lipases and pseudopyronine B for the degradation of oil

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Abstract

The lipA gene, encoding a solvent-tolerant extracellular lipase from Proteus sp. SW1, was displayed on the cell surface of Escherichia coli by fusing it to an antigen 43 anchoring motif. The display of LipA on the Escherichia coli cell surface was directly confirmed by immunofluorescence microscopy and flow cytometry. After 6 days of incubation in media containing 1 % used cooking oil, an Escherichia coli strain expressing surface displayed lipase was able to degrade 27 % of the oil. The biosurfactant, pseudopyronine B, was purified from culture supernatants of Pseudomonas sp. SL31. Its critical micelle concentration was determined to be 1400 mg/l, and the surfactant was stable within a temperature range from 0 to 120 °C and a pH range of 3–11. Pseudopyronine B-containing crude media extracts efficiently removed up to 51 % of the cadmium from contaminated water. We demonstrated the oil degradation ability of the mixed culture of four bacterial strains, namely the recombinant Escherichia coli expressing cell surface displayed lipase (pKKJlipA), His-tagged lipase (pETlipA), extracellular lipase-producing Proteus sp. SW1, and pseudopyronine B-producing Pseudomonas sp. SL31 by culturing in LB media containing 1 % oil. The consortium degraded 29 % of oil in one day and reached 84 % after 7 days.

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Acknowledgments

This research work was supported in part by a grant from the Chulabhorn Research Institute and Center of Excellence on Environmental Health and Toxicology, Thailand. Special thanks go to Piyajit Watcharasit for Texas red-conjugated anti-rabbit IgG antibody, Nuchanart Rangkadilok and Supachai Ritruechai for preparative HPLC, Amnart Khongmanee for his kind assistance with fluorescence microscopes, James Dubbs for critical reading and discussion, Pakwilai Chouichit for technical assistance, and also to Piyapol Munpiyamit for photograph preparation. We appreciate the funding by the Royal Golden Jubilee Ph.D. Program (PHD/0144/2554) to Sirinthra Thiengmag.

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Authors and Affiliations

  1. Applied Biological Sciences Program, Chulabhorn Graduate Institute, Bangkok, 10210, Thailand

    S. Thiengmag, S. Chuencharoen, S. Mongkolsuk & S. Loprasert

  2. Chemical Biology Program, Chulabhorn Graduate Institute, Bangkok, 10210, Thailand

    N. Thasana

  3. Laboratory of Medicinal Chemistry, Chulabhorn Research Institute, Bangkok, 10210, Thailand

    N. Thasana

  4. Laboratory of Biotechnology, Chulabhorn Research Institute, Kamphaengphet 6 Rd., Bangkok, 10210, Thailand

    W. Whangsuk, S. Mongkolsuk & S. Loprasert

  5. Department of Chemical Engineering, Faculty of Engineering, Burapha University, Chonburi, 20131, Thailand

    W. Jangiam

  6. Center of Excellence on Environmental Health and Toxicology, Ministry of Education, Bangkok, 10400, Thailand

    S. Mongkolsuk & S. Loprasert

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  1. S. Thiengmag
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Correspondence to S. Loprasert.

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Thiengmag, S., Chuencharoen, S., Thasana, N. et al. Bacterial consortium expressing surface displayed, intra- and extracellular lipases and pseudopyronine B for the degradation of oil. Int. J. Environ. Sci. Technol. 13, 2067–2078 (2016). https://doi.org/10.1007/s13762-016-1034-z

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  • DOI: https://doi.org/10.1007/s13762-016-1034-z

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