Analysis of a Sulfur-oxidizing Perchlorate-degrading Microbial Community

황 산화를 통해 퍼클로레이트를 분해하는 미생물 군집 분석

  • Received : 2015.08.15
  • Accepted : 2015.10.07
  • Published : 2016.01.30


Perchlorate (ClO4) is an emerging pollutant detected in surface water, soil, and groundwater. Previous studies provided experimental evidence of autotrophic ClO4 removal with elemental sulfur (S0) particles and activated sludge, which are inexpensive and easily available, respectively. In addition, ClO4 removal efficiency was shown to increase when an enrichment culture was used as an inoculum instead of activated sludge. PCR-DGGE was employed in the present study to investigate the microbial community in the enrichment culture that removed ClO4 autotrophically. Microorganisms in the enrichment culture showed 99.71% or more ClO4 removal efficiency after a 7-day incubation when the initial concentration was approximately 120 mg ClO4/l. Genomic DNA was isolated from the enriched culture and its inoculum (activated sludge), and used for PCR-DGGE analysis of 16S rRNA genes. Microbial compositions of the enrichment culture and the activated sludge were different, as determined by their different DGGE profiles. The difference in DGGE banding patterns suggests that environmental conditions of the enrichment culture caused a change in the microbial community composition of the inoculated activated sludge. Dominant DGGE bands in the enrichment culture sample were affiliated with the classes β-Proteobacteria, Bacteroidetes, and Spirochaetes. Further investigation is warranted to reveal the metabolic roles of the dominant populations in the ClO4 degradation process, along with their isolation.


Bacterial community;enrichment culture;elemental sulfur;perchlorate;PCR-DGGE


  1. Ahn, Y., Park, E. J., Oh, Y. K., Park, S., Webster, G. and Weightman, A. J. 2005. Biofilm microbial community of a thermophilic trickling biofilter used for continuous biohydrogen production. FEMS Microbiol. Lett. 249, 31-38.
  2. Altschul, S. F., Madden, T. I., Schifer, A. J., Zhang, J., Zhang, Z., Miller, W. and Lipman, D. J. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25, 3389-3402.
  3. Bardiya, N. and Bae, J. H. 2011. Dissimilatory perchlorate reduction: a review. Microbiol. Res. 166, 237-254.
  4. Coates, J. D. and Achenbach, L. A. 2004. Microbial perchlorate reduction: Rocket-fuelled metabolism. Nat. Rev. Microbiol. 2, 569-580.
  5. Kim, H., Kim, J. and Lee, Y. 2007. Occurrence of perchlorate in drinking water in Korea. J. Kor. Soc. Water Quality 23, 822-828.
  6. Kim, H., Kim, J., Lee, Y., Lee, J. and Kim, S. 2008. Perchlorate in advanced drinking water treatment process. J. Kor. Soc. Water Quality 24, 164-168.
  7. Larkin, M. A., Blackshields, G., Brown, N. P., Chenna, R., McGettigan, P. A., McWilliam, H., Valentin, F., Wallace, I. M., Wilm, A., Lopez, R., Thompson, J. D., Gibson T. J. and Higgins, D. G. 2007. Clustal W and Clustal X version 2. Bioinformatics 23, 2947-2948.
  8. Lee, C. 2009. Optimum treatment of sewage and wastewater discharged in Gumi industrial complex. Final report 09-2-10-16-5. Gyeongbuk regional environment technology development center. Gyeongbuk, Korea.
  9. Karavaiko, G. I., Dubinina, G. A. and Kondrat’eva, T. F. 2006. Lithotrophic microorganisms of the oxidative cycles of sulfur and iron. Microbiology 75, 512-545.
  10. US EPA. 1999. EPA METHOD 314.0: Determination of perchlorate in drinking water using ion chromatography.
  11. Sahu, A. K., Conneely, T., Nüsslein, K. R. and Ergas, S. J. 2009. Biological perchlorate reduction in packed bed reactors using elemental sulfur. Environ. Sci. Technol. 43, 4466-4471.
  12. Shin, K. H., Son, A., Cha, D. K. and Kim, K. W. 2007. Review on risks of perchlorate and treatment technologies. J. Kor. Soc. Environ. Eng. 29, 1060-1068.
  13. Timura, K., Stecher, G., Peterson, D., Filipski, A. and Kumar, S. 2013. MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol. 30, 2725-2729.
  14. Feng, Y., Xu, Y., Yu, Y., Xie, Z. and Lin, X. 2012. Mechanisms of biochar decreasing methane emission from Chinese paddy soils. Soil Biol. Biochem. 46, 80-88.
  15. Gao, M., Wang, S., Jin, C., She, Z., Zhao, C., Zhao, Y., Zhang, J. and Ren, Y. 2015. Autotrophic perchlorate reduction kinetics of a microbial consortium using elemental sulfur as an electron donor. Environ. Sci. Pollut. Res. 22, 9694-9703.
  16. Han, K. R., Kang, T. H., Kang, H. C., Kim, K., Seo, D. H. and Ahn, Y. 2011. Autotrophic perchlorate-removal using elemental sulfur granules and activated sludge: batch test. J. Life Sci. 21, 1473-1480.
  17. Han, K. R. and Ahn, Y. 2013. Characterization of perchlorate-removal using elemental sulfur granules and activated sludge. J. Life Sci. 23, 676-681.
  18. Republic of Korea Ministry of Environment. 2010. Guideline for the management of drinking water quality monitoring items.
  19. Merlino, G., Rizzi, A., Schievano, A., Tenca, A., Scaglia, B., Oberti, R., Adani, F. and Daffonchio, D. 2013. Microbial community structure and dynamics in two-stage vs single-stage thermophilic anaerobic digestion of mixed swine slurry and market biowaste. Water Res. 47, 1983-1995.
  20. Motzer, W. E. 2001. Perchlorate: problems, detection, and solutions. Environ. Forensics 2, 301-311.
  21. Nocker, A., Burr, M. and Burr, A. K. 2007. Genotypic microbial community profiling: a critical technical Review. Microbiol. Ecol. 54, 276-289.