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Enhanced detoxification of organophosphates using recombinant Escherichia coli with co-expression of organophosphorus hydrolase and bacterial hemoglobin

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Abstract

A significantly improved, recombinant Escherichia coli has been developed to degrade the toxic organophosphorus compound, Paraoxon, to non-toxic materials by co-expression of organophosphorus hydrolase (OPH) under trc promoter and Vitreoscilla hemoglobin (VHb) under O2dependent nar promoter. VHb-expressing whole cells had significant enhancement of OPH activity (48%, 18.7 vs. 27.8 unit l−1) and bioconversion efficiency V max/K m (44%, 0.14 vs. 0.2 min−1) compared to VHb-free system.

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References

  • Caldwell SR, Newcomb JR, Schlecht KA, Raushel FM (1991) Limits of diffusion in the hydrolysis of substrates by the phophotriesterase from Pseudomonas diminuta. Biochemistry 30: 7438-7444.

    Google Scholar 

  • Cha HJ, Wu CF, Valdes JJ, Rao G, Bentley WE (2000) Observations of green fluorescent protein as a fusion partner in genetically engineered Escherichia coli: monitoring protein expression and solubility. Biotechnol. Bioeng. 67: 565-574.

    Google Scholar 

  • Chen W, Mulchandani A (1998) The use of live biocatalysts for pesticide detoxification. Trends Biotechnol. 16: 71-76.

    Google Scholar 

  • Dikshit KL, Webster DA (1988) Cloning, characterization and expression of the bacterial hemoglobin gene from Vitreoscilla in Escherichia coli. Gene 70: 377-386.

    Google Scholar 

  • Dikshit KL, Kikshit RP, Webster DA (1990) Study of Vitreoscilla hemoglobin (vhb) gene expression and promoter activity in E. coli through transcriptional fusion. Nucl. Acids Res. 18: 4149-4155.

    Google Scholar 

  • Donarski WJ, Dumas DP, Heitmeyer DP, Lewis VE, Raushel FM (1989) Structure-activity relationship in the hydrolysis of substrates by the phosphotriesterase from Pseudomonas diminuta. Biochemistry 28: 4650-4655.

    Google Scholar 

  • Grimsley JK, Scholtz JM, Pace CN, Wild JR (1997) Organophosphorus hydrolase is a remarkably stable enzyme that unfolds through a homodimeric intermediate. Biochemistry 36: 14366-14374.

    Google Scholar 

  • Khosla C, Bailey JE (1988) The Vitreoscilla hemoglobin gene: molecular cloning, nucleotide sequence and genetic expression in Escherichia coli. Mol. Gen. Genet. 214: 158-161.

    Google Scholar 

  • Khosla C, Bailey JE (1989) Characterization of the oxygendependent promoter of the Vitreoscilla hemoglobin gene in Escherichia coli. J. Bacteriol. 171: 5995-6004.

    Google Scholar 

  • Lai K, Stolowich NJ, Wild JR (1995) Characterization of P-S bond hydrolysis in organophosphorothioate pesticides by organophosphorus hydrolase. Arch. Biochem. Biophys. 318: 59-64.

    Google Scholar 

  • Lewis VE, Donarski WJ, Wild JR, Raushel FM (1988) Mechanism and stereochemical course at phosphorus of the reaction catalyzed by a bacterial phosphotriesterase. Biochemistry 27: 1591-1597.

    Google Scholar 

  • Li SF, DeMoss JA (1987) Promoter region of the nar operon of Escherichia coli: nucleotide sequence and transcription initiation signals. J. Bacteriol. 169: 4614-4620.

    Google Scholar 

  • Liu SC, Webster DA, Wei M, Stark BC (1996) Genetic engineering to contain the Vitreoscilla hemoglobin gene enhances degradation of benzoic acid by Xanthomonas maltophilia. Biotechnol. Bioeng. 49: 101-105.

    Google Scholar 

  • Mansee AH, Chen W, Mulchandani A (2000) Biodetoxification of coumaphos insecticide using immobilized Escherichia coli expressing organophosphorus hydrolase enzyme on cell surface. Biotechnol. Bioprocess Eng. 5: 436-440.

    Google Scholar 

  • McDaniel CS, Harper LL, Wild JR (1988) Cloning and sequencing of a plasmid-borne gene (opd) encoding a phosphotriesterase. J. Bacteriol. 170: 2306-2311.

    Google Scholar 

  • Mulbry WW, Karns JS (1989) Parathion hydrolase specified by the Flavobacterium opd gene: relationship between the gene and protein. J. Bacteriol. 171: 6740-6746.

    Google Scholar 

  • Patel SM, Stark BC, Hwang KW, Dikshit KL, Webster DA (2000) Cloning and expression of Vitreoscilla hemoglobin gene in Bukholderia sp. strain DNT for enhancement of 2,4-dinitrotoluene degradation. Biotechnol. Prog. 16: 26-30.

    Google Scholar 

  • Seong GH, Han SJ, Chang HN, Lee J (1997) Whole cell enzyme microencapsulation of Escherichia coli with oxygen-dependent inducible nar promoter. Biotechnol. Lett. 19: 881-884.

    Google Scholar 

  • Serdar CM, Gibson DT (1985) Enzymatic hydrolysis of organophosphates: cloning and expression of a parathion hydrolase gene from Pseudomonas diminuta. Bio/Technology 3: 567-571.

    Google Scholar 

  • Shuler ML, Kargi F (1992) Bioprocess Engineering: Basic Concepts. New Jersey: Prentice Hall.

    Google Scholar 

  • Wu CF, Valdes JJ, Rao G, Bentley WE (2001) Enhancement of organophosphorus hydrolase yield in Escherichia coli using multiple gene fusions. Biotechnol. Bioeng. 75: 100-103.

    Google Scholar 

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Authors and Affiliations

  1. Department of Chemical Engineering & Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang, 790-784, Korea

    Dong Gyun Kang, Jaoon Y.H. Kim & Hyung Joon Cha

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  1. Dong Gyun Kang
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  2. Jaoon Y.H. Kim
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  3. Hyung Joon Cha
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Kang, D.G., Kim, J.Y. & Cha, H.J. Enhanced detoxification of organophosphates using recombinant Escherichia coli with co-expression of organophosphorus hydrolase and bacterial hemoglobin. Biotechnology Letters 24, 879–883 (2002). https://doi.org/10.1023/A:1015568727825

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