Biotransformation of the Fungicide Chlorothalonil by Bacterial Glutathione S-Transferase

  • Kim, Young-Mog (Institute of Agricultural Science and Technology, Kyungpook National University) ;
  • Park, Kun-Bawui (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Choi, Jun-Ho (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Kim, Jang-Eok (Department of Agricultural Chemistry, Kyungpook National University) ;
  • Rhee, In-Koo (Department of Agricultural Chemistry, Kyungpook National University)
  • Published : 2004.10.01

Abstract

A gene responsible for the chlorothalonil-biotransformation was cloned from the chromosomal DNA of Ochrobactrum anthropi SH35B, an isolated bacterium strain from soil. We determined the nucleotide sequences and found an open reading frame for glutathione S-transferase (GST). The drug-hypersensitive Escherichia coli KAM3 cells transformed with a plasmid carrying the GST gene can grow in the presence of chlorothalonil. The GST of O. anthropi SH35B was expressed in E. coli and purified by affinity chromatography. The fungicide chlorothalonil was rapidly transformed by the purified GST in the presence of glutathione. No significant difference in the chlorothalonil-biotransformation effect was observed among the thiol compounds (cysteine, reduced glutathione, and $\beta$-mercaptoethanol). Thus, the result reported here is the first evidence on the chlorothalonil-biotransformation by conjugation with the cellular free thiol groups, especially glutathione, catalyzed by the bacterial GST.

Keywords

References

  1. Berns, K. I. and C. A. Thomas. 1965. Isolation of the high molecular DNA from Haemophilus influenzae. J. Mol. Biol. 11: 476- 490.
  2. Blom, A., W. Harder, and A. Martin. 1992. Unique and overlapping pollutant stress proteins of Escherichia coli. Appl. Environ. Microbial. 58: 331- 334.
  3. Bradford, M. M. 1976. A rapid and sensitive method for the quantitification of microgram quantities of protein utilizing the principle of protein dye binding. Anal. Biochem. 72: 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  4. Di llio, C, A. Aceto, R. Piccolornini, N. Allocati, A. Faraone, L. Cellini, G. Ravagnan, and G. Federici. 1988. Purification and characterization of three forms of glutathione transferase from Proteus mirabilis. Biochem. J. 255: 971- 975.
  5. Di Ilio, C., A. Aceto, R. Piccolomini, N. Allocati, A. Faraone, T Bucciarelli, D. Barra, and G. Federici. 1991. Purification and characterization of a novel glutathione transferase from Serratia marcescens. Biochim. Biophys. Acta 1077: 141-146.
  6. Di Ilio, C., A. Aceto, N. Allocati, R. Piccolomini, T Bucciarelli, B. Dragani, A. Faraone, P. Sacchetta, R. Petruzzelli, and G. Federici. 1993. Characterization of glutathione transferase from Xanthomonas campestris. Arch. Biochem. Biophys. 305: 110- 114.
  7. Dunbar, J., L. O. Ticknor, and C. R. Kuske. 2000. Assessment of microbial diversity in four Southwestern United States soils by 16S rRNA gene terminal restriction fragment analysis. Appl. Environ. Microbiol. 66: 2943-2950.
  8. Favaloro, B., A. Tamburro, S. Angelucci, A. D. Luca, S. Melino, C. Dillio, and D. Rotilio. 1998. Molecular cloning, expression and site-directed mutagenesis of glutathione Stransferase from Ochrobactrum anthropi. Biochem. J. 335: 573- 579.
  9. Jakobi, W. B., W. H. Habig, J. H. Keen, J. N. Ketley, and M. J. Pabst. 1976. Glutathione S-transferases: Catalytic aspects, pp. 189- 211. In: Glutathione: Metabolism and Function. Raven Press, New York, U.S.A.
  10. Hamer, D. 1986. Metallothionein. Annu. Rev. Biochem. 55: 913- 951.
  11. Huh, N. E., N. S. Choi, Y K. Seo, T S. Yu, and H. S. Lee. 1994. Characterization of cadmium-resistant yeast strain in response to cadmium or heat shock stress. J. Microbiol. Biotechnol. 4: 30- 35.
  12. Inouhe, M., M. Hiyama, H. Tohoyama, M. Joho, and T Murayama. 1989. Cadmium-binding protein in a cadmiumresistant strain of Saccharomyces cerevisiae. Biochim. Biophys. Acta 993: 51- 55.
  13. Kimura, N., A. Nishi, M. Goto, and K. Furukawa. 1997. Functional analyses of a variety of chimeric dioxygenases constructed from two biphenyl dioxygenases that are similar structurally but different functionally. J. Bacteriol. 179: 3936- 3943.
  14. Kwon, H. H., E. Y Lee, K. S. Cho, and H. W. Ryu. 2003. Benzene biodegradation using the polyurethane biofilter immobilized with Stenotrophomonas maltophilla T3-c. J. Microbiol. Biotechnol. 13: 70- 76.
  15. Leavitt, J. R. C. and D. Penner. 1979. In vitro conjugation of glutathione and other thiols with acetanilide herbicides and EPTC sulfoxide and the action of the herbicide antidote R25788.J. Agric. Food Chem. 27: 533- 536.
  16. Lee, S. H., J. H. Shin, J. H. Choi, J. W. Park, J. E. Kim, and I. K. Rhee. 2004. Isolation and characterization of chlorothalonildissipating bacteria from soil. Kor. J. Microbiol. Biotechnol. 32: 96-100.
  17. Lee, S. K. and S. B. Lee. 2002. Substrate utilization patterns during BTEX biodegradation by an o-xylene-degrading bacterium Ralstonia sp. PHS1. J. Microbiol. Biotechnol. 12: 909-915.
  18. Lindquist, S. and E. A. Craig. 1988. The heat-shock proteins. Annu. Rev. Genet. 22: 631-677.
  19. Meister, A. 1985. Methods for the selective modification of glutathione metabolism and study of glutathione transport. Methods Enzymol. 113: 571- 583.
  20. Meister, A. and M. E. Anderson. 1983. Glutathione. Annu. Rev. Biochem. 52: 711- 760.
  21. Morita, Y, K. Kodama, S. Shiota, T. Mine, A. Kataoka, T. Mizushima, and T. Tsuchiya. 1998. NorM, a putative multidrug efflux protein, of Vibrio parahaemolyticus and its homolog in Escherichia coli. Antimicrob. Agents Chemother. 42: 1778- 1782.
  22. Nishida, M., K. H. Kong, H. Inoue, and K. Takahashi. 1994. Molecular cloning and site-directed mutagenesis of glutathione S-transferase from Escherichia coli. The conserved tyrosyl residue near the N terminus is not essential for catalysis. J. BioI. Chem. 269: 32536- 32541.
  23. Park, M. K., K. H. Liu, Y H. Lim, Y H. Lee, H. G. Hur, and J. H. Kim. 2003. Biotransformation of a fungicide ethaboxam by soil fungus Cunninghamella elegans. J. Microbiol. Biotechnol. 13: 43- 47.
  24. Paulsen, I. T, R. Seshadri, K. E. Nelson, J. A. Eisen, J. F. Heidelberg, T D. Read, R. J. Dodson, L. Umayam, L. M. Brinkac, M. J. Beanan, S. C. Daugherty, R. T Deboy, A. S. Durkin, J. F. Kolonay, R. Madupu, W. C. Nelson, B. Ayodeji, M. Kraul, J. Shetty, J. Malek, S. E. Van Aken, S. Riedmuller, H. Tettelin, S. R. Gill, O. White, S. L. Salzberg, D. L. Hoover, L. E. Lindler, S. M. Halling, S. M. Boyle, and C. M. Fraser. 2002. The Brucella suis genome reveals fundamental similarities between animal and plant pathogens and symbionts. Proc. Natl. Acad. Sci. USA 99: 13148-13153.儀돀⢸ᒗ⨀큨?⨀җ⨀㣏ᒗ⨀넃돐恾⨀恾⨀褃돀邰ᒗ⨀岝䨄傻ᒗ⨀傻ᒗ⨀Ȁ怀炻ᒗ⨀炻ᒗ⨀肻ᒗ⨀肻ᒗ⨀㢷ꎶয়Ѐ̀Ѐᄔ᐀Ą耇잖⨀䬀￿
  25. Reha-Krantz, L. J. 1985. The Escherichia coli strain JM105 contains partial supE activity. Gene 38: 275- 276. https://doi.org/10.1016/0378-1119(85)90229-X
  26. Sambrook, J., E. F. Fritch, and T. Maniatis. 1989. Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y, U.S.A.
  27. Sanger, F., S. Nicklen, and A. R. Coulson. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463- 5467. https://doi.org/10.1073/pnas.74.12.5463
  28. Scarponi, L., P. Perucci, and L. Martinetti. 1991. Conjugation of 2-chloroacetanilide herbicides with glutathione: Role of molecular structures and of glutathione S-transferase enzymes. J. Agric. Food Chem. 39: 2010-2013.
  29. Schlesinger, M. J. 1990. Heat shock proteins. J. Biol. Chem. 265: 12111-12114.
  30. Sheehan, D. and J. P. Casey. 1993. Microbial glutathione Stransferases. Compo Biochem. Physiol. B. 104: 1- 6.
  31. Shin, J. H., Y M. Kim, J. W. Park, J. E. Kim, and 1.K. Rhee. 2003. Resistance of Saccharomyces cerevisiae to fungicide chlorothalonil. J. Microbiol. 43: 219- 223.
  32. Wilce, M. C. J. and M. W. Parker. 1994. Structure and function of glutathione S-transferase. Biochim. Biophys. Acta 1205: 1-18.