Journal of Korean Society of Environmental Engineers (대한환경공학회지)

Korean Society of Environmental Engineers (대한환경공학회)
권호 표지

Characterization of Denitrifier Community in Independent Anoxic Reactor Using Modified BAF Process

Modified BAF 공정을 이용한 독립적인 무산소조에서 탈질미생물 군집의 특성

  • Park, Jeung-Jin (Department of Environmental Engineering, Pusan National University) ;
  • Jeung, Young-Rok (Department of Environmental Engineering, Pusan National University) ;
  • Yu, Jae-Cheul (Department of Environmental Engineering, Pusan National University) ;
  • Hur, Sung-Ho (Department of Environmental Engineering, Pusan National University) ;
  • Choi, Won-Seok (Bioentech company, Ltd.) ;
  • Byun, Im-Gyu (Institute for Environmental Technology and Industry, Pusan National University) ;
  • Lee, Tae-Ho (Department of Environmental Engineering, Andong National University) ;
  • Park, Tae-Joo (Department of Environmental Engineering, Pusan National University)
  • 박정진 (부산대학교 환경공학과) ;
  • 정영록 (부산대학교 환경공학과) ;
  • 유재철 (부산대학교 환경공학과) ;
  • 허성호 (부산대학교 환경공학과) ;
  • 최원석 ((주)바이오엔텍) ;
  • 변임규 (부산대학교 환경기술산업개발연구센터) ;
  • 이태호 (안동대학교 환경공학과) ;
  • 박태주 (부산대학교 환경공학과)
  • Published : 2006.07.31
Download PDF
⟨ Previous Next ⟩

Abstract

The independent anoxic reactor was introduced in biological aerated filters as the regulation of water quality requirement, especially total nitrogen, had been strengthened. The process studied in this work was upflow $Biobead^{(R)}$ process which was used commercial invented for removal of organic materials and nitrification. For the purpose of evaluating the independent anoxic reactor, PCR-DGGE, of the molecular biological methods, was performed. Two types of nitrite reductase genes were selected. One is nirS represented cytocrome $cd_1$ nitrite reductase gene and the other is nirK represented Cu-containing nitrite reductase gene. Denitrifier community in the independent anoxic reactor was analyzed with PCR-DGGE using these two denitrifying functional genes. As the result of the PCR, only nirS gene was detected between nirS and nirK. With the result of the DGGE, specific bands became strong, as the operating days were longer, nitrate loading rate was increased. otherwise those of the initial activated sludge showed various bands. In the consequence of the sequence of DGGE bands, various denitrifiers were sequenced in the initial activated sludge, while specific denitrifiers like alcaligenes faecalis were predominant in the anoxic reactor. Consequently, introduction of the independent anoxic reactor made it possible to achieve 96% denitrification efficiency, and was proper for the modification of BAF process.

최근 수계의 총질소(T-N) 규제가 강화되면서 기존 BAF 공정의 개선을 위해 독립적인 무산소조가 추가로 도입되었다. 본 연구에 사용된 공정은 유기물과 질산화 중심으로 개발 된 기술인 $Biobead^{(R)}$공법으로 상용화된 상향류의 BAF공정의 하나이다. 독립적인 무산소조의 도입의 타당성을 검토하기 위해 분자생물학적 방법의 하나인 PCR-DGGE기법이 수행되었다. 두 가지 type의 nitrite reductase genes를 통해 진행되었는데, nirS로 암호화된 cytocrome $cd_1$ nitrite reductase gene과 nirK로 암호화된 Cu를 함유한 nitrite reductase gene이다. 이러한 탈질 기능유전자를 이용하여 PCR-DGGE를 통해 탈질 목적으로 순화된 독립적인 무산소조의 탈질미생물의 군집을 해석하였다. PCR 증폭결과, 탈질을 수행하는 무산소조 내에서는 nirS와 nirK유전자 가운데 nirS유전자만 검출되었고, DGGE 분석결과, 최초 식종원으로 이용된 활성슬러지에서는 상대적으로 많은 band들이 검출되는 반면, 무산소조 내에서는 운전일수와 nitrate 부하량이 증가할수록 단일 band로 우점화 하는 경향을 나타내었다. DGGE band에 대한 염기서열 분석결과, 식종 슬러지의 경우 다양한 uncultured bacteria가 나타났으나, nitrate 제거율이 높은 안정화된 무산소조에서는 alcaligenes faecalis 등 특정 탈질미생물이 우점화 되는 것으로 확인되었다. 결론적으로 이러한 탈질미생물 군집특성을 가지는 무산소조의 도입은 96%이상의 안정적인 탈질을 가능하게 하였으며, BAF 공정 개선을 위한 독립적인 무산소조의 도입은 적절한 것으로 판단되었다.

Keywords

References

  1. Yukio, I. and Akihiro, T., 'Nitrogen Removal from Municipal Wastewater by a Single Submerged Filter,' JWPCF, 56, 251-258(1984)
  2. Frank, R., 'Nitrification and Phosphorus Precipitation with Biological Aerated Filters,' JWPCF, 62, 169-176(1992)
  3. Tom, S., Allan, M., and John, U., 'The small Footprint Wastewater Treatment Process,' Chemistry & Industry, 19, 533-536(1993)
  4. Tschui, M., 'Tertiary Nitrification in Aerated Pilot Biofilters,' Water Sci. Technol., 29(10-11), 53-60(1994)
  5. 건설신기술 제289호, http://www.bioentech.com
  6. Barnes, D. and Bliss, P. J., 'Biological control of nitrogen in wastewater treatment,' University Press, Cambridge, UK(1983)
  7. Zumft, W. G., 'Cell biology and molecular basis of denitrification,' Microbiol. Mol. Bio. R., 61, 533-616 (1997)
  8. Ye, R. W., Averill, B. A., Tiedje, J. M., 'Denitrification : Production and Consumption of Nitric Oxide,' Appl. Environ. Microb., 60(4), 1053-1058(1994)
  9. Brader, G., Zhou, J., Wu, L., Devol, A. H., and Tiedje, J. M., 'Nitrite reductase genes(NirK and NirS) as functional markers to investigate diversity of denitrifying bacteria in pacific northwest marine sediment communities,' Appl. Environ. Microb., 66, 2096-2104(2000) https://doi.org/10.1128/AEM.66.5.2096-2104.2000
  10. 환경부, 수질오염공정시험방법(2004)
  11. APHA, 'Standard methods for the examination of water and wastewater,' American Public Health Association, Washington, D.C.(1995)
  12. Braker, G., Fesefeldt, A., Witzel, K. P., 'Development of PCR primer systems for amplification of nitrite reductase genes(nirK and nirS) to detect denitrifying bacteria in environmental samples,' Appl. Environ. Microb., 64, 3769-3775(1998)
  13. Michotey, V., Mejean, V., Bonin, P., 'Comparison of methods for quantification of cytochrome cdl-denitrifying bacteria in marine samples,' Appl. Environ. Microb., 66, 1564-1571(2000) https://doi.org/10.1128/AEM.66.4.1564-1571.2000
  14. Throback, I. N., Enwall, K, Jarvis, A., Hallin, S., 'Reassessing PCR primers targeting nirS, nirK and nosZ genes for community surveys of denitrifying bacteria with DGGE,' FEMS Microbiol. Ecol., 49(3), 401-417(2004) https://doi.org/10.1016/j.femsec.2004.04.011
  15. Casciotti, K. L., Ward, B. B., 'Dissimilatory nitrite reductase genes from autotrophic ammonia-oxdizing bacteria,' Appl. Environ. Microb., 67, 2213-2221(2001) https://doi.org/10.1128/AEM.67.5.2213-2221.2001
  16. Song, B. K and Ward, B. B., 'Nitrate reductase genes in halobenzoate degrading denitrifying bacteria,' FEMS Microbiol. Ecol., 43, 349-357(2003) https://doi.org/10.1111/j.1574-6941.2003.tb01075.x
  17. Martin Wolsing and Anders Prieme, 'Obsevation of high seasonal variation in community structure of denitrifying bacteria in arable soil receiving artificial fertilizer and cattle manure by determining T-RFLP of nir gene fragments,' FEMS Microbiol. Ecol., 48, 261-271(2004) https://doi.org/10.1016/j.femsec.2004.02.002
  18. Gesche Braker, Andreas Fesefeldt, Kar-Paul Witzel, 'Development of PCR primer systems for amplification of nitrite reductase genes(nirS and nirK) to detect denitrifying bacteria in environmental samples,' Applied and Environmental Microbiology, 64, 3769-3775(1998)
  19. Cho, S. J., 'Analysis of nitrifying bacterial communities and characteristics of aerobic denitrifiers in biological nitrogen removal,' Ph.D. thesis, Department of Microbiology, Pusan national university, Pusan, South Korea(2006)