Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Analysis of Archaeal Communities in Full-Scale Anaerobic Digesters Using 454 Pyrosequencing

454 Pyrosequencing을 이용한 실규모 혐기성 소화조의 아케아 군집구조 분석

  • Kang, Hyun-Jin (School of Civil, Environmental and Architectural Engineering, Korea University) ;
  • Kim, Taek-Seung (School of Civil, Environmental and Architectural Engineering, Korea University) ;
  • Lee, Young-Haeng (Water Research Center, Korea Institute of Science and Technology) ;
  • Lee, Taek-June (Water Research Center, Korea Institute of Science and Technology) ;
  • Han, Keum-Suk (Waterworks Research Institute, Seoul Metropolitan Government) ;
  • Choi, Young-Jun (Waterworks Research Institute, Seoul Metropolitan Government) ;
  • Park, Hee-Deung (School of Civil, Environmental and Architectural Engineering, Korea University)
  • 강현진 (고려대학교 건축사회환경공학부) ;
  • 김택승 (고려대학교 건축사회환경공학부) ;
  • 이영행 (한국과학기술연구원 물연구센터) ;
  • 이택준 (한국과학기술연구원 물연구센터) ;
  • 한금석 (서울특별시 상수도연구원) ;
  • 최영준 (서울특별시 상수도연구원) ;
  • 박희등 (고려대학교 건축사회환경공학부)
  • Received : 2011.06.16
  • Accepted : 2011.09.01
  • Published : 2011.09.30
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Abstract

Archaeal communities were investigated using 454 pyrosequencing technology based on 16S rRNA gene in 11 samples collected from six different full-scale anaerobic digesters. Observed operational taxonomic units (OTUs) estimated from the archaeal 16S rRNA gene sequences were 13-55 OTUs (3% cutoff) which was corresponded to 29-89% of Chao1 richness estimates. In the anaerobic digesters there were archaeal sequences within the orders Thermoproteales, Thermoplasmatales, Desulfurococcales as well as within the orders Methanomicrobiales, Methanobacteriales, Methanococcales, Methanosarcinales, and Methanocellales, which are known to produce methane. Among these orders, Methanococcales known to produce methane using hydrogen was the predominant taxon and constituted 51.8-99.7% of total sequences. All samples showed a very similar community structure (Pearson correlation coefficient=0.99) except for one sample based on a heat map analysis. In addition, canonical correspondence analysis correlating archaeal communities to the environmental variables demonstrated that digester temperature and total solids removal rate were the two important explanatory variables. Overall results suggested that environmental and operational variables of anaerobic digester are important factors determining archaeal diversity and community structure.

결론적으로 본 연구에서는 16S rRNA 유전자 기반의 454 pyrosequencing을 통해 혐기성 소화조에 존재하는 다양한 아케아를 규명할 수 있었으며, 지금까지 그 중요성이 잘 알려지지 않은 Methanococcales 목에 속하는 아케아가 소화조에 공통적으로 존재함을 확인할 수 있었다. 또한, 소화조의 운전조건은 아케아의 다양성과 군집구조를 결정하는데 영향을 미치는 중요한 인자라는 것을 알 수 있었다. 실규모 6개 혐기성 소화조를 대상으로 11개 소화 슬러지 시료를 채취해, 16S rRNA 유전자 기반의 454 pyrosequencing을 이용하여 아케아 군집구조를 조사하였다. 아케아 16S rRNA 유전자 염기서열로부터 측정된 observed operational taxanomic units (OTUs)는 13-55 OTUs이었으며(3% cutoff), 이는 Chao1 richness estimate로 계산된 값의 29-89%에 해당하였다. 소화조에는 메탄생성에 직접적으로 관련이 있다고 알려진 Methanomicrobiales, Methanobacteriales, Methanococcales, Methanosarcinales, Methanocellales 목에 속하는 아케아 뿐만 아니라, Thermoproteales, Thermoplasmatales, Desulfurococcales목에 속하는 아케아도 함께 발견되었다. 이 중 수소를 기질로 해서 메탄을 생성한다고 알려진 Methanoacoccales가 전체 염기서열의 51.8-99.7%를 차지해 가장 많이 발견된 분류군으로 나타났다. Heat map 분석결과 각 시료들의 아케아 군집구조는 1개 시료를 제외한 나머지 10개 시료가 매우 비슷한 군집구조를 가지고 있었다(Pearson 상관지수=0.99). 또한, 아케아 군집구조와 환경변수들의 상관성을 분석한 canonical correspondence analysis를 통해 혐기성 소화조의 아케아 군집구조에 영향을 미치는 중요 환경변수는 소화조의 온도와 총고형물 제거율임을 확인하였다. 모든 결과들을 종합해 볼 때 소화조의 운전조건은 아케아의 다양성과 군집구조를 결정하는데 영향을 미치는 중요한 인자라고 사료된다.

Keywords

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