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Simultaneous degradation of organophosphate and organochlorine pesticides by Sphingobium japonicum UT26 with surface-displayed organophosphorus hydrolase

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Abstract

A genetically engineered microorganism (GEM) capable of simultaneously degrading organophosphate and organochlorine pesticides was constructed for the first time by display of organophosphorus hydrolase (OPH) on the cell surface of a hexachlorocyclohexane (HCH)-degrading Sphingobium japonicum UT26. The GEM could potentially be used for removing the two classes of pesticides that may be present in mixtures at contaminated sites. A surface anchor system derived from the truncated ice nucleation protein (INPNC) from Pseudomonas syringae was used to target OPH onto the cell surface of UT26, reducing the potential substrate uptake limitation. The surface localization of INPNC–OPH fusion was verified by cell fractionation, western blot, proteinase accessibility, and immunofluorescence microscopy. Furthermore, the functionality of the surface-exposed OPH was demonstrated by OPH activity assays. Surface display of INPNC–OPH fusion (82 kDa) neither inhibited cell growth nor affected cell viability. The engineered UT26 could degrade parathion as well as γ-HCH rapidly in minimal salt medium. The removal of parathion and γ-HCH by engineered UT26 in sterile and non-sterile soil was also studied. In both soil samples, a mixture of parathion (100 mg kg−1) and γ-HCH (10 mg kg−1) could be degraded completely within 15 days. Soil treatment results indicated that the engineered UT26 is a promising multifunctional bacterium that could be used for the bioremediation of multiple pesticide-contaminated environments.

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Acknowledgments

We thank Yuji Nagata of Tohoku University, Japan, for providing Sphingobium japonicum UT26. This work was supported by grants from Agriculture Research Program of Science and Technology Department of Liaoning Province, China (Nos. 2011211001 and 2009209001).

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Authors and Affiliations

  1. School of Life Science, Liaoning University, Shenyang, 110036, China

    Xiangyu Cao & Jianli Liu

  2. Department of Microbiology, College of Life Science, Nankai University, Tianjin, 300071, China

    Chao Yang, Qiang Li, Wei Zhang & Cunjiang Song

  3. State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin, 300071, China

    Ruihua Liu

  4. State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China

    Chuanling Qiao

  5. Department of Chemical and Environmental Engineering, University of California, Riverside, Riverside, CA, 92521, USA

    Ashok Mulchandani

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  1. Xiangyu Cao
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  2. Chao Yang
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  4. Qiang Li
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  5. Wei Zhang
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  8. Chuanling Qiao
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  9. Ashok Mulchandani
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Correspondence to Chao Yang.

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Cao, X., Yang, C., Liu, R. et al. Simultaneous degradation of organophosphate and organochlorine pesticides by Sphingobium japonicum UT26 with surface-displayed organophosphorus hydrolase. Biodegradation 24, 295–303 (2013). https://doi.org/10.1007/s10532-012-9587-0

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  • DOI: https://doi.org/10.1007/s10532-012-9587-0

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