Korean Journal of Soil Science and Fertilizer (한국토양비료학회지)

Korean Society of Soil Science and Fertilizer (한국토양비료학회)
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Isolation and Characterization of Sulfur-oxidizing Denitrifying Bacteria Utilizing Thiosulfate as an Electron Donor

황(thiosulfate)을 이용하는 탈질 미생물의 분리 및 특성 파악

  • Oh, Sang-Eun (Division of Biological Environment, Kangwon National University) ;
  • Joo, Jin-Ho (Division of Biological Environment, Kangwon National University) ;
  • Yang, Jae E (Division of Biological Environment, Kangwon National University)
  • Received : 2006.01.21
  • Accepted : 2006.01.06
  • Published : 2006.12.30
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Abstract

Sulfur-oxidizing bacteria were enumerated and isolated from a steady-state anaerobic master culture reactor (MCR) operated for over six months under a semi-continuous mode and nitrate-limiting conditions using thiosulfate as an electron donor. Most are Gram-negative bacteria, which have sizes up to 12 m. Strains AD1 and AD2 were isolated from the plate count agar (PCA), and strains BD1 and BD2 from the solid thiosulfate/nitrate medium. Based on the morphological, physiological, FAME and 16S rDNA sequence analyses, the two dominant strains, AD1 and AD2, were identified as Paracoccus denitrificans and Paracoccus versutus (formerly Thiobacillus versutus), respectively. From the physiological results, glucose was assimilated by both strains AD1 and AD2. Heterotrophic growth of strains AD1 and AD2 could be a more efficient way of obtaining a greater amount of biomass for use as an inoculum. Even though facultative autotrophic bacteria grow under heterotrophic conditions, autotrophic denitrification would not be reduced.

황산화 미생물인 Genus Thiobacillus 중 몇 종류의 탈질균은 여러 종류의 황 화합물($S^{2-}$, So, $S_2O{_3}^{2-}$, $S_4O{_6}^{2-}$, $SO{_3}^{2-}$)을 황산염이온으로 산화시키면서 동시에 질산성질소를 질소 가스 형태로 전환시킨다. 이는 독립영양 미생물이므로 외부 탄소원이 필요치 않으며, C/N비가 낮은 폐수에 에탄올 대신 값이 싼 황 입자의 투입으로 경제적이며 효과적인 탈질화를 유도할 수 있다. 후탈질시 인위적인 유기물의 투입대신 값 싼 황입자를 사용하여 질소를 제거하며, 처리효율이 안정적이고, 운전이 쉬워, 최근 이 공정은 세계적으로 많이 연구되고 있다. 그러나 탈질시 알칼리도가 파괴되어 특히 알칼리도가 낮은 폐수의 경우 고농도 탈질시 pH가 떨어져 탈질이 더 이상 진행되지 않으며, 고농도의 질산성질소를 처리할 경우 부산물로서 고농도의 황산염이온이 생성된다는 부수적인 단점을 안고 있다. 본 연구에서는 MCR로부터 독립영양 미생물을 분리 characterization 및 미생물 동정을 하였다. MCR내에는 주로 Paracoccus denitrificans and Paracoccus versutus (formerly Thiobacillus versutus)가 주종을 이루었으며 이들 미생물들은 유기물도 에너지원으로 이용할 수 있는 facultative autotrophic denitrifier이었다. 이들 미생물을 탈질에 이용할 시 유기물이 있는 조건에서도 성장하기 때문에 유입 폐수 내 유기물 농도에 영향을 받지 않고 또는 인위적으로 소량의 유기물을 넣어 독립영양탈질과 종속영양탈질을 동시에 일어날 수 있도록 함으로서 독립영양탈질의 단점인 높은 황산염이온 생성 및 알칼리도 파괴를 막을 수 있을 것으로 사료된다.

Keywords

Acknowledgement

Supported by : Kangwon National University

References

  1. Atlas, R.M., and R. Bartha. 1993. Microbial ecology. 3rd ed., The Benjamin/Cummings publishing co., USA
  2. Batchelor, B., and A.W. Lawrence. 1978. Autotrophic denitrification using elemental sulfur. Journal WPCF. 50:1986-2001
  3. Claus G., and H.J. Kutzner. 1985. Autotrophic denitrification by Thiobacillus denitrificans in a packed bed reactor. Appl Microbiol Biotechnol. 22:289-296
  4. Felsenstein, J. 1993. PHYLIP (Phylogenetic Inference Package) version 3.5C. Distributed by the author. Department of Genetics, University of Washington, Seattle, USA
  5. Flere, J.M., and T.C. Zhang. 1999. Nitrate removal with sulfur-limestone autotrophic denitrification processes. J of Envir. Engrg. 8(125):721-729
  6. Holt, J.G., N.R Krieg, P.H.A. Sneath, J.T. Staley, and S.T. Williams. 1994. Bergey's Manual of Determinative Bacteriology. 9th Edn. Williams & Wilkins, Baltimore, USA
  7. Koenig, A., and L.H. Liu. 1996. Autotrophic denitrification of landfill leachate using elemental sulphur. Wat. Sci. Tech. 34(5-6):469-476
  8. Kuenen, J.G. 1979. Growth yields and maintenance energy requirement in Thiobacillus species under energy limitation. Arch. Microbiol. 122:183-188 https://doi.org/10.1007/BF00411358
  9. Matin, A. 1978. Organic Nutrition of Chemolithotrophic Bacteria. Ann. Rev. Microbiol. 32:433-68 https://doi.org/10.1146/annurev.mi.32.100178.002245
  10. Matsui, S., and R.Yamamoto, 1986. A new method of sulphur denitrification for sewage treatment by a fluidized bed reactor. Wat. Sci. Tech. 18:355-362
  11. Oh, S.-E, K.-S. Kim, H.-C. Choi, J. Cho, and I.S. Kim. 2000. Kinetics and physiology of autotrophic denitrification by denitrifying sulfur bacteria. Wat. Sci. Tech. 42(3-4):59-68
  12. Page, R.D.M. 1996. Treeview: an application to display phylogenetic trees on personal computers. Comput. Appl. Biosci. 12:357-358
  13. Thompson, J.D., T.J. Gibson, F. Plewniak, F. Jeanmougin, and D.G. Higgins. 1997. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 24:4876-4882
  14. Yoon, J.H., S.T. Lee, and Y.H. Park. 1998. Inter- and intraspecific phylogenetic analysis of the genus Nocardioidesand related taxa based on 16S rDNA sequences. Int. J. Syst. Bacteriol. 48:187-194 https://doi.org/10.1099/00207713-48-1-187
  15. Zhang, T.C., and D.G. Lampe. 1999. Sulfur:Limestone autotrophic denitrification processes for treatment of nitrate-contaminated water: Batch experiments. Wat. Res. 33(3):599-608 https://doi.org/10.1016/S0043-1354(98)00281-4