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Biodegradation

, Volume 21, Issue 6, pp 903–913 | Cite as

Screening for and isolation and identification of malathion-degrading bacteria: cloning and sequencing a gene that potentially encodes the malathion-degrading enzyme, carboxylestrase in soil bacteria

  • Sayed K. Goda
  • Iman E. Elsayed
  • Taha A. Khodair
  • Walaa El-Sayed
  • Mervat E. Mohamed
Original Paper

Abstract

Five malathion-degrading bacterial strains were enriched and isolated from soil samples collected from different agricultural sites in Cairo, Egypt. Malathion was used as a sole source of carbon (50 mg/l) to enumerate malathion degraders, which were designated as IS1, IS2, IS3, IS4, and IS5. They were identified, based on their morphological and biochemical characteristics, as Pseudomonas sp., Pseudomonas putida, Micrococcus lylae, Pseudomonas aureofaciens, and Acetobacter liquefaciens, respectively. IS1 and IS2, which showed the highest degrading activity, were selected for further identification by partial sequence analysis of their 16S rRNA genes. The 16S rRNA gene of IS1 shared 99% similarity with that of Alphaprotoebacterium BAL284, while IS2 scored 100% similarity with that of Pseudomonas putida 32zhy. Malathion residues almost completely disappeared within 6 days of incubation in IS2 liquid cultures. LC/ESI-MS analysis confirmed the degradation of malathion to malathion monocarboxylic and dicarboxylic acids, which formed as a result of carboxylesterase activity. A carboxylesterase gene (CE) was amplified from the IS2 genome by using specifically designed PCR primers. The sequence analysis showed a significant similarity to a known CE gene in different Pseudomonas sp. We report here the isolation of a new malathion-degrading bacteria from soils in Egypt that may be very well adapted to the climatic and environmental conditions of the country. We also report the partial cloning of a new CE gene. Due to their high biodegradation activity, the bacteria isolated from this work merit further study as potential biological agents for the remediation of soil, water, or crops contaminated with the pesticide malathion.

Keywords

Malathion Pesticide degradation Pseudomonas GC-ECD Carboxylesterase 

Notes

Acknowledgement

The author would like to thank Shafallah Medical Genetic Centre (SMGC), Doha, Qatar for the DNA sequencing and other technical support. The author also would like to thank Yaseen I Gad for his help and support.

References

  1. Bhadhade BJ, Sarnaik SS, Kanekar PP (2002) Bioremediation of an industrial effluent containing monocrotophos. Curr Microbiol. 45:346–349CrossRefGoogle Scholar
  2. Cáceres T, He W, Naidu R, Megharaj M (2007) Toxicity of chlorpyrifos and TCP alone and in combination to Daphnia carinata: the influence of microbial degradation in natural water. Water Res 41:4497–4503CrossRefPubMedGoogle Scholar
  3. Chapalamadugu S, Chaudhry GR (1992) Microbiological and biotechnological aspects of metabolism of carbamates and organophosphates. Crit Rev Biotechnol 12:357–389CrossRefPubMedGoogle Scholar
  4. Ellman GL, Courtney KD, Andres V Jr, Featherstone RH (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95CrossRefPubMedGoogle Scholar
  5. Feng Y, Racke KD, Bollag JM (1997) Isolation and characterization of a chlorinated-pyridinol-degrading bacterium. Appl Environ Microbiol 63:4096–4409PubMedGoogle Scholar
  6. Galli E (1994) The role of microorganism in the environment decontamination. In: Renzoni A, Mattei N, Lari L, Fossi MC (eds) Contaminants in the environment—a multidisciplinary assessment of risks to man and other organisms. CRC Press, Boca Raton, pp 235–296Google Scholar
  7. George MG, Julia AB, Timothy L (2005) Betaproteobacteria. In: Brenner DJ, Krieg NR, Staley JT (eds) Bergey’s manual of systematic bacteriology, 2nd edn. Springer, New York, pp 575–600Google Scholar
  8. Gosselin RE (1984) Clinical toxicology of commercial products. Williams and Wilkins, BaltimoreGoogle Scholar
  9. Jakoby WE, Ziegler DM (1990) The enzymes of detoxification. J Biol Chem 265:20715–20718PubMedGoogle Scholar
  10. Kakugawa S, Fushinobu F, Wakagi T, Shoun H (2007) Characterization of a thermostable carboxylesterase from the hyperthermophilic bacterium Thermotoga maritime. Appl Microb Biotechnol 74:585–591CrossRefGoogle Scholar
  11. Kim HE, Lee IS, Kim JH, Hahn KW, Park VJ, Han HS, Park KR (2003) Gene cloning, sequencing, and expression of an esterase from Acinetobacter lwoffii I6C-l. Curr Microbiol 46:291–295CrossRefPubMedGoogle Scholar
  12. Kim YH, Ahn JY, Moon SH, Lee J (2005) Biodegradation and detoxification of organophosphate insecticide, malathion by Fusarium oxysporum f. sp. pisi cutinase. Chemosphere 60:1349–1355CrossRefPubMedGoogle Scholar
  13. Lane DJ (1991) 16S/23S rRNA sequence. In: Stackebrandt M, Goodfellow M (eds) Nucleic acid techniques in bacterial systematic. Wiley, New York, pp 115–175Google Scholar
  14. Laveglia J, Dahm PA (1977) Degradation of organophosphorus and carbamate insecticides in the soil and by soil microorganisms. Annu Rev Entomol 22:483–513CrossRefPubMedGoogle Scholar
  15. Lewis DL, Paris DF, Baughman GL (1975) Transformation of malathion by a fungus, Aspergillus oryzae, isolated from a fresh water pond. Bull Environ Contam Toxicol 13:596–601CrossRefPubMedGoogle Scholar
  16. Li X, He J, Li S (2007) Isolation of a chlorpyrifos-degrading bacterium, Sphingomonas sp. strain Dsp-2, and cloning of the mpd gene. Res Microbiol 158(2):14–143CrossRefGoogle Scholar
  17. Merone L, Mandrich L, Rossi M, Manco G (2005) A thermostable phosphotriesterase from the archaeon Sulfolobus solfataricus: cloning, overexpression and properties. Extremophiles 9:297–305CrossRefPubMedGoogle Scholar
  18. Parekh NR, Walker A, Roberts SJ, Welch SJ (1994) Rapid degradation of triazinone herbicide metmitron by a Rhodococcus sp. Isolated from treated soil. J Appl Bacteriol 77:467–475PubMedGoogle Scholar
  19. Paris DF, Lewis DL, Wolfe NL (1975) Rates of degradation of malathion by bacteria isolated from aquatic systems. Environ Sci Technol 9:135–138CrossRefGoogle Scholar
  20. Pimentel ID (1983) Effects of pesticides on the environment. In Proceedings of the 10th international congress on plant protection, vol 2. Crydon, UK, pp 685–691Google Scholar
  21. Racke KD, Coats RJ (1987) Enhanced degradation of isofenphos by soil microorganisms. J Agric Food Chem 35:94CrossRefGoogle Scholar
  22. Racke KD, Laskowski DA, Schultz MR (1990) Resistance of chlorpyrifos to enhanced biodegradation in soil, J. Agric Food Chem 38:1430–1436CrossRefGoogle Scholar
  23. Saaty RP, Booth SR (1994) In situ bioremediation: Cost effectiveness of a remediation technology field tested at the Savannach river integrated demonstration site. LA-UR-94-1714. Los Alamos National Laboratory, Los Alamos, New MexicoGoogle Scholar
  24. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, vol 3. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  25. Singh AK, Seth PK (1989) Degradation of malathion by microorganisms isolated from industrial effluents. Bull Environ Contam Toxicol 43:28–35CrossRefPubMedGoogle Scholar
  26. Singh BK, Walker A (2006) Microbial degradation of organophosphorus compounds. FEMS Microbiol Rev 30:428–471CrossRefPubMedGoogle Scholar
  27. U.S. EPA. Office of Pesticide Programs (1988) Pesticides in ground water data base: interim report. Washington, DCGoogle Scholar
  28. Uygun U, Özkara R, Özbey A, Koksel H (2007) Residue levels of malathion and fenitrothion and their metabolites in post harvest treated barley during storage and malting. Food Chem 100:1165–1169CrossRefGoogle Scholar
  29. Walker WW (1976) Chemical and microbial degradation of malathion and parathion in an estuarine environment. J Environ Qual 5:210–216CrossRefGoogle Scholar
  30. WHO (1997) The WHO recommended classification of pesticides by hazard and guidelines to classification, 1996–1997. WHO, Geneva, 64 ppGoogle Scholar
  31. Yoshii K, Tonogai Y, Katakawa J, Ueno H, Nakamuro K (2007) Kinetic analysis for hydrolysis of malathion by carboxylesterase in wheat karnels. J Health Sci 53:507–513CrossRefGoogle Scholar
  32. Zhang J, Lan W, Qiao C, Jiang H (2004) Bioremediation of organophosphorus pesticides by surface-expressed carboxylesterase from mosquito on Escherichia coli. Biotechnol Prog 20:1567–1571CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Sayed K. Goda
    • 1
    • 2
  • Iman E. Elsayed
    • 2
  • Taha A. Khodair
    • 3
  • Walaa El-Sayed
    • 3
  • Mervat E. Mohamed
    • 2
  1. 1.Shafallah Medical Genetic CentreDohaQatar
  2. 2.Faculty of ScienceCairo UniversityCairoEgypt
  3. 3.Faculty of AgricultureAin Shams UniversityCairoEgypt

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