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Isolation of a novel gene encoding a 3,5,6-trichloro-2-pyridinol degrading enzyme from a cow rumen metagenomic library

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Abstract

3,5,6-trichloro-2-pyridinol (TCP) is a major metabolite of the insecticide chlorpyrifos and is hazardous to human and animal health. A gene encoding a TCP degrading enzyme was cloned from a metagenomic library prepared from cow rumen. The gene (tcp3A) is 2.5 kb in length, encoding a protein (Tcp3A) of 599 amino acid residues. Tcp3A has a potential signal sequence, as well as a putative ATP/GTP binding site, and a likely amidation site. The molecular weight of the enzyme is 62 kDa by SDS–PAGE. Comparison of Tcp3A with the NCBI database using BLASTP revealed homology to amidohydrolase proteins. Recombinant Escherichia coli harboring the tcp3A gene could utilize TCP as the sole source of carbon. TLC and HPLC revealed that TCP was degraded by recombinant E. coli harboring tcp3A. This is the first report of a gene encoding a TCP degrading enzyme.

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Acknowledgments

This work was supported by a Grant No. R01-2008-000-20220-0 from the Basic Research Program of KOSEF, Korea. Renukaradhya K. Math and Shah Md. Asraful Islam are supported by scholarships from the BK21 Program, Ministry of Education & Human Resources Development, Korea. We are very much thankful to BioScience Writers, LCC, 8418 Bluegate St. 713-664-4597, Houston, TX 77025 for the correction of our manuscript.

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Authors and Affiliations

  1. Division of Applied Life Science (BK21 Program), Gyeongsang National University, Chinju, 660-701, Republic of Korea

    Renukaradhya K. Math, Shah Md. Asraful Islam, Sun Joo Hong, Jong Min Kim, Myoung Geun Yun, Ji Joong Cho & Han Dae Yun

  2. Life Science, Gyeongsang National University, Chinju, 660-701, Republic of Korea

    Han Dae Yun

  3. Department of Food Science, Jinju National University, Chinju, 660-758, Republic of Korea

    Kye Man Cho

  4. Division of Plant Environmental Research, Gyeongsangnam-do Agricultural Research and Extension Service, Chinju, 660-360, Republic of Korea

    Jae Young Heo & Young Han Lee

  5. Department of Agricultural Chemistry, Sunchon National University, Suncheon, 540-742, Republic of Korea

    Hoon Kim

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  1. Renukaradhya K. Math
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  2. Shah Md. Asraful Islam
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  3. Kye Man Cho
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  4. Sun Joo Hong
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  5. Jong Min Kim
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Correspondence to Han Dae Yun.

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Renukaradhya K. Math, and Shah Md. Asraful Islam contributed equally in this work.

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Math, R.K., Asraful Islam, S.M., Cho, K.M. et al. Isolation of a novel gene encoding a 3,5,6-trichloro-2-pyridinol degrading enzyme from a cow rumen metagenomic library. Biodegradation 21, 565–573 (2010). https://doi.org/10.1007/s10532-009-9324-5

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  • DOI: https://doi.org/10.1007/s10532-009-9324-5

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