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Applied Microbiology and Biotechnology

, Volume 93, Issue 3, pp 1305–1314 | Cite as

Characterization of a marine-isolated mercury-resistant Pseudomonas putida strain SP1 and its potential application in marine mercury reduction

  • Weiwei Zhang
  • Lingxin ChenEmail author
  • Dongyan Liu
Environmental Biotechnology

Abstract

The Pseudomonas putida strain SP1 was isolated from marine environment and was found to be resistant to 280 μM HgCl2. SP1 was also highly resistant to other metals, including CdCl2, CoCl2, CrCl3, CuCl2, PbCl2, and ZnSO4, and the antibiotics ampicillin (Ap), kanamycin (Kn), chloramphenicol (Cm), and tetracycline (Tc). mer operon, possessed by most mercury-resistant bacteria, and other diverse types of resistant determinants were all located on the bacterial chromosome. Cold vapor atomic absorption spectrometry and a volatilization test indicated that the isolated P. putida SP1 was able to volatilize almost 100% of the total mercury it was exposed to and could potentially be used for bioremediation in marine environments. The optimal pH for the growth of P. putida SP1 in the presence of HgCl2 and the removal of HgCl2 by P. putida SP1 was between 8.0 and 9.0, whereas the optimal pH for the expression of merA, the mercuric reductase enzyme in mer operon that reduces reactive Hg2+ to volatile and relatively inert monoatomic Hg0 vapor, was around 5.0. LD50 of P. putida SP1 to flounder and turbot was 1.5 × 109 CFU. Biofilm developed by P. putida SP1 was 1- to 3-fold lower than biofilm developed by an aquatic pathogen Pseudomonas fluorescens TSS. The results of this study indicate that P. putida SP1 is a low virulence strain that can potentially be applied in the bioremediation of HgCl2 contamination over a broad range of pH.

Keywords

Pseudomonas putida Marine environment mer operon Bioremediation of HgCl2 contamination 

Notes

Acknowledgments

We sincerely thank Dr. L. Sun of the Institute of Oceanology, Chinese Academy of Sciences and her laboratory for kind help in providing the bacterial strain P. fluorescens TSS. This work was financially supported by Innovation Projects of the Chinese Academy of Sciences grant KZCX2-EW-206 and KZCX2-YW-Q07-04, the National Natural Science Foundation of China (NSFC) grant 20975089, and the 100 Talents Program of the Chinese Academy of Sciences.

References

  1. Ahn MC, Kim B, Holsen TM, Yi SM, Han YJ (2010) Factors influencing concentrations of dissolved gaseous mercury (DGM) and total mercury (TM) in an artificial reservoir. Environ Pollut 158:347–355CrossRefGoogle Scholar
  2. Altinok I, Kayis S, Capkin E (2006) Pseudomonas putida infection in rainbow trout. Aquaculture 261:850–855CrossRefGoogle Scholar
  3. Bafana A, Krishnamurthi K, Patil M, Chakrabarti T (2010) Heavy metal resistance in Arthrobacter ramosus strain G2 isolated from mercuric salt-contaminated soil. J Hazard Mater 177:481–486CrossRefGoogle Scholar
  4. Barkay T, Wagner-Döbler I (2005) Microbial transformations of mercury: potentials, challenges, and achievements in controlling mercury toxicity in the environment. Adv Appl Microbiol 57:1–52CrossRefGoogle Scholar
  5. Barkay T, Miller SM, Summers AO (2003) Bacterial mercury resistance from atoms to ecosystems. FEMS Microbiol Rev 27:355–384CrossRefGoogle Scholar
  6. Bouallèguea O, Mzoughia R, Weillc FX, Mahdhaouib N, Salema YB, Sbouib H, Grimontc F, Grimont PAD (2004) Outbreak of Pseudomonas putida bacteraemia in a neonatal intensive care unit. J Hosp Infect 57:88–91CrossRefGoogle Scholar
  7. De J, Ramaiah N (2007) Characterization of marine bacteria highly resistant to mercury exhibiting multiple resistances to toxic chemicals. Ecol Indic 7:511–520CrossRefGoogle Scholar
  8. De J, Ramaiah N, Mesquita A, Verlekar XN (2003) Tolerance to various toxicants by marine bacteria highly resistant to mercury. Mar Biotechnol 5:185–193CrossRefGoogle Scholar
  9. Dietz R, Outridge PM, Hobson KA (2009) Anthropogenic contributions to mercury levels in present-day Arctic animals—a review. Sci Total Environ 407:6120–6131CrossRefGoogle Scholar
  10. Golding GR, Kelly CA, Sparling R, Loewen PC, Rudd JWM, Barkay T (2002) Evidence for facilitated uptake of Hg(II) by Vibrio anguillarum and Escherichia coli under anaerobic and aerobic conditions. Limnol Oceanogr 47:967–975CrossRefGoogle Scholar
  11. Golding G, Sparling R, Kelly C (2008) Effect of pH on intracellular accumulation of trace concentrations of Hg(II) in Escherichia coli under anaerobic conditions, as measured using a mer–lux bioreporter. Appl Environ Microbiol 74:667–675CrossRefGoogle Scholar
  12. Guzzi G, La Porta CA (2008) Molecular mechanisms triggered by mercury. Toxicology 244:1–12CrossRefGoogle Scholar
  13. Hansen CL, Zwolinski G, Martin D, Williams JW (1984) Bacterial removal of mercury from sewage. Biotechnol Bioeng 26:1330–1333CrossRefGoogle Scholar
  14. Hideomi N, Ishikawa T, Yasunaga S, Kondo I, Mitsuhasi S (1977) Frequency of heavy-metal resistance in bacteria from inpatients in Japan. Nature 266:165–167CrossRefGoogle Scholar
  15. Hu YH, Liu CS, Hou JH, Sun L (2009) Identification, characterization, and molecular application of a virulence-associated autotransporter from a pathogenic Pseudomonas fluorescens strain. Appl Environ Microbiol 75:4333–4340CrossRefGoogle Scholar
  16. Kado CI, Liu ST (1981) Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol 145:1365–1373Google Scholar
  17. Kannan SK, Krishnamoorthy R (2006) Isolation of mercury resistant bacteria and influence of abiotic factors on bioavailability of mercury—a case study in Pulicat Lake North of Chennai, South East India. Sci Total Environ 367:341–353CrossRefGoogle Scholar
  18. Kelly CA, Rudd JWM, Holoka MH (2003) Effect of pH on mercury uptake by an aquatic bacterium: implications for Hg cycling. Environ Sci Technol 37:2941–2946CrossRefGoogle Scholar
  19. Kholodii G, Bogdanova E (2002) Tn5044-conferred mercury resistance depends on temperature: the complexity of the character of thermosensitivity. Genetica 115:233–241CrossRefGoogle Scholar
  20. Kholodii G, Yurieva V, Gorlenko Zh, Mindlin S, Bass I, Lomovskaya O, Kopteva AV, Nikiforov G (1997) Tn5047: a chimeric mercury resistance transposon closely related to the toluene degradative transposon Tn4657. Microbiology 143:2549–2556CrossRefGoogle Scholar
  21. Kholodii G, Yurieva O, Mindlin S, Gorlenko Z, Rybochkin V, Nikiforov V (2000) Tn5044, a novel Tn3 family transposon coding for temperature-sensitive mercury resistance. Res Microbiol 151:291–302CrossRefGoogle Scholar
  22. Lane D, Pace B, Olsen G, Stahl D, Sogin M, Pace N (1985) Rapid determination of 16S ribosomal sequences for phylogenetic analyses. Proc Natl Acad Sci USA 82:6955–6959CrossRefGoogle Scholar
  23. Li P, Feng XB, Qiu GL, Shang LH, Li ZG (2009) Mercury pollution in Asia: a review of the contaminated sites. J Hazard Mater 168:591–601CrossRefGoogle Scholar
  24. Mindlin S, Kholodii G, Gorlenko Z, Minakhina S, Minakhin L, Kalyaeva E, Kopteva A, Petrova M, Yurieva O, Nikiforov V (2001) Mercury resistance transposons of Gram-negative environmental bacteria and their classification. Res Microbiol 152:811–822CrossRefGoogle Scholar
  25. Mindlin SZ, Bass IA, Bogdanova ES, Gorlenko ZM, Kalyaeva ES, Petrova MA, Nikiforov VG (2002) Horizontal transfer of mercury resistance genes in environmental bacterial populations. Mol Biol 36:160–170CrossRefGoogle Scholar
  26. Mindlin S, Minakhin L, Petrova M, Kholodii G, Minakhina S, Gorlenko Z, Nikiforov V (2005) Present-day mercury resistance transposons are common in bacteria preserved in permafrost grounds since the Upper Pleistocene. Res Microbiol 156:994–1004CrossRefGoogle Scholar
  27. Mirzaei N, Kafilzadeh F, Kargar M (2008) Isolation and identification of mercury resistant bacteria from Kor River, Iran. J Biol Sci 8:935–939CrossRefGoogle Scholar
  28. Mortazavi S, Rezaee A, Khavanin A, Varmazyar S, Jafarzadeh M (2005) Removal of mercuric chloride by a mercury resistant Pseudomonas putida strain. J Biol Sci 5:269–273CrossRefGoogle Scholar
  29. Murtaza I, Dutt A, Ali A (2002) Relationship between the persistence of mer operon sequences in Escherichia coli and their resistance to mercury. Curr Microbiol 44:178–183CrossRefGoogle Scholar
  30. Nakamura K, Nakahara H (1988) Simplified X-ray film method for detection of bacterial volatilization of mercury chloride by Escherichia coli. Appl Environ Microbiol 54:2871–2873Google Scholar
  31. Nascimento AMA, Chartone-Souza E (2003) Operon mer: bacterial resistance to mercury and potential for bioremediation of contaminated environments. Genet Mol Res 2:92–101Google Scholar
  32. Oehmen A, Fradinho J, Serra S, Carvalho G, Capelo JL, Velizarov S, Crespo JG, Reis MAM (2009) The effect of carbon source on the biological reduction of ionic mercury. J Hazard Mater 165:1040–1048CrossRefGoogle Scholar
  33. Osborn AM, Bruce KD, Strike P, Ritchie DA (1997) Distribution, diversity and evolution of the bacterial mercury resistance (mer) operon. FEMS Microbiol Rev 19:239–262CrossRefGoogle Scholar
  34. Parsek MR, Singh PK (2003) Bacterial biofilms: an emerging link to disease pathogenesis. Annu Rev Microbiol 57:677–701CrossRefGoogle Scholar
  35. Partridge SR, Brown HJ, Stokes HW (2001) Transposons Tn1696 and Tn21 and their integrons In4 and In2 have independent origins. Antimicrob Agents Chemother 45:1263–1270CrossRefGoogle Scholar
  36. Pepi M, Gaggi C, Bernardini E, Focardi S, Lobianco A, Ruta M, Nicolardi V, Volterrani M, Gasperini S, Trinchera G, Renzi P, Gabellini M, Focardi SE (2010) Mercury-resistant bacterial strains Pseudomonas and Psychrobacter spp. isolated from sediments of Orbetello Lagoon (Italy) and their possible use in bioremediation processes. Int Biodeter Biodegr 65:85–91CrossRefGoogle Scholar
  37. Poulain AJ, Ní Chadhain SM, Ariya PA, Amyot M, Garcia E, Campbell PG, Zylstra GJ, Barkay T (2007) Potential for mercury reduction by microbes in the high arctic. Appl Environ Microbiol 73:2230–2238CrossRefGoogle Scholar
  38. Singh S, Kang SH, Mulchandani A, Chen W (2008) Bioremediation: environmental clean-up through pathway engineering. Curr Opin Biotechnol 19:437–444CrossRefGoogle Scholar
  39. Syn CK, Swarup S (2000) A scalable protocol for isolation of large-sized genomic DNA with in an hour from several bacteria. Anal Biochem 278:86–90CrossRefGoogle Scholar
  40. Von Canstein H, Li Y, Timmis KN, Deckwer WD, Wagner-Döbler I (1999) Removal of mercury from chloralkali electrolysis wastewater by a mercury-resistant Pseudomonas putida strain. Appl Environ Microbiol 65:5279–5284Google Scholar
  41. Wagner-Döbler I (2003) Pilot plant for bioremediation of mercury-containing industrial wastewater. Appl Microbiol Biotechnol 62:124–133CrossRefGoogle Scholar
  42. Wang HR, Hu YH, Zhang WW, Sun L (2009) Construction of an attenuated Pseudomonas fluorescens strain and evaluation of its potential as a cross-protective vaccine. Vaccine 27:4047–4055CrossRefGoogle Scholar
  43. Xu L, Li H, Vuong C, Vadyvaloo V, Wang J, Yao Y, Otto M, Gao Q (2006) Role of the luxS quorum-sensing system in biofilm formation and virulence of Staphylococcus epidermidis. Infect Immun 74:488–496CrossRefGoogle Scholar
  44. Zhang WW, Sun L (2007) Cloning, characterization, and molecular application of a beta-agarase gene from Vibrio sp. strain V134. Appl Environ Microbiol 73:2825–2831CrossRefGoogle Scholar
  45. Zhang WW, Sun K, Cheng S, Sun L (2008) Characterization of DegQVh, a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain. Appl Environ Microbiol 74:6254–6262CrossRefGoogle Scholar
  46. Zhang WW, Hu YH, Wang HL, Sun L (2009) Identification and characterization of a virulence-associated protease from a pathogenic Pseudomonas fluorescens strain. Vet Microbiol 139:183–188CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Yantai Institute of Coastal Zone ResearchChinese Academy of SciencesYantaiChina
  2. 2.Key Laboratory of Coastal Zone Environmental Processes, Chinese Academy of Sciences; Shandong Provincial Key Laboratory of Coastal Zone Environmental Processes, Yantai Institute of Coastal Zone ResearchChinese Academy of SciencesYantaiChina

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