Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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PCR Detection of Terephthalic Acid Degrading Comamonas testosteroni in Soil

PCR을 이용한 토양 중 Terephthalic Acid 분해 Comamonas testosteroni의 검출

  • 이종훈 (경기대학교 식품생물공학과) ;
  • 강동주 (경기대학교 식품생물공학과) ;
  • 홍연표 (중앙대학교 예방의학교실)
  • Published : 2003.06.01
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Abstract

Eleven bacterial strains which are able to utilize terephthalic acid as a carbon and an energy source for growth were isolated from the soil of 7 water quality evaluation points in Kyonggi area of Korea. Phthalic acid isomer degrading activity of the isolates from the 4 contaminated points was higher than those from the 3 clean points. Among 11 isolates, 4 isolates which have high terephthalic acid degrading activity and degrade two phthalic acid isomers were identified by partal 16S rDNA sequence determination. One of them was identified as Pseudomonas putida, and the others as Comamonas testosteroni. Thus a large number of phthalic acid isomer degrading bacteria in domestic soil were inferred as C. testosteroni. On the basis of these results, the PCR detection of C. testosteroni in soil was applied to monitor soil contamination by phthalic acid isomers. The DNA of C. test-osteroni extracted from 4 g soil was directly detected by PCR with C. testosteroni specific primer pair. The amount of PCR products was different according to sampling sites and more PCR products were obtained from contaminated sites than those from clean sites (Gulpo-chun＞Anyang-chun＞Hwangguji-chun>Shin-chun＞Huk-chun＞Pukhan-river>Kapyeong-chun). This result was coincided with that of the viable cell counts for terephthalic acid degrading bacteria.

경기도 내의 7개 하천 토양으로부터 terephthalic acid를 분해, 대사하는 미생물 11개 균주를 분리하고, 이들의 phthalic acid 이성질체 분해활성을 비교한 결과, 오염도가 높은 4개 지역에서 분리된 균들의 phthalic acid 이성질체 분해활성이 오염도가 낮은 3개 지역에서 분리된 균들에 비해 높은 것으로 나타났다. 분리된 균주들 중, 높은 terephthalic acid 분해활성을 보유하고 2개의 phthalic acid 이성질체의 분해가 가능한 4균주의 165 rDNA 부분 염기서열결정을 통하여 국내 토양에서 생육하고 있는 phthalic acid이성질체 분해미생물 중 많은 수가 C. testosteroni인 것으로 추정되었다. 이러한 결과에 근거하여 토양 중 C. testosteroni의 특이적 검출에 의한 phthalic acid 이성질체에 의한 토양오염의 모니터링 가능성을 검토하였다. C. testosteroni 특이적 PCR primer를 구축하고 토양으로부터 직접 추출한 DNA를 template로 PCR을 수행한 결과, 각 하천 주변 4 g의 토양으로부터 관찰 가능한 정도의 PCR산물을 얻을 수 있었다. 증폭된 PCR산물의 양은 굴포천＞안양천＞황구지천＞신천＞흑천＞북한강＞가평천의 순으로 청정지역에서 보다 오염지역의 토양에서 C. testosteroni가 많이 생육하고 있는 것으로 나타났으며, 각 지역의 토양에 존재하는 terephthalic acid 분해균의 생균수 또한 PCR에 의한 C. testosteroni의 검출과 동일한 양상으로 나타났다.

Keywords

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