• 제목/요약/키워드: meta-cleavage

검색결과 51건 처리시간 0.023초

대장균에서 분리된 din (damage-inducible)과 tin (temperature-inducible) 유전자들의 특성 (Characterization of the din (damage-inducible) and tin (temperature-inducible) Genes Isolated from Escherichia coli)

  • 백경희
    • 미생물학회지
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    • 제29권6호
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    • pp.392-396
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    • 1991
  • Pseudomonas sp. DJ77의 chromosomal DNA로부터 6.9kb XhoI 절편 상에 존재하는 phenanthrene 분해에 관련된 유전자군을 vector pBLUESCRIPT SK(+)에 클로닝하였다. 이렇게 얻은 재조합 plasmid인 pHENX7을 가지고 있는 JM101 균주는 3-methylcarechol을 노란색의 meta-cleavage 화합물로 전환할 수 있었다. 그러나 삽입된 절편의 방향이 반대가 되도록 제조한 pHENX7은 extradiol dioxygenase 활성을 나타내지 않기 때문에 전사방향을 알 수 있었다. pHENX7과 이의 듀도체들을 지니는 JH101균주에서 PhnC(24kDa), PhnD(31KDa), PhnE(34kDa), PhnF(KDa)의 4 polypeptide를 확인 할 수 있었고 개개의 유전자의 위치와 범위를 알 수 있었다. 유전자 순서는 phnC-phnD-phnE-phnF-phnG이었으며, phnC, phnD, phnE, phnF, phnG는 각기 glutathione S-transferase, meta-cleavage compound hydrolase extradiol dioxygenase, meta-cleavage compound dehydrogenase의 유전자이었다.

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Construction of a Bioluminescent Reporter Using the luc Gene and meta-Cleavage Dioxygenase Promoter for Detection of Catecholic Compounds

  • Park, Sang-Ho;Lee, Dong-Hun;Oh, Kye-Heon;Kim, Chi-Kyung
    • Journal of Microbiology
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    • 제38권3호
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    • pp.183-186
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    • 2000
  • Several types of bioluminescent reporter strains have been developed for the detection and monitoring of pollutant aromatics contaminating the environment. In this study, a bioluminescent reporter strain, E. coli SHP3, was constructed by fusing the luc gene of firefly luciferase with the promoter of pcbC responsible for the meta-cleavage of aromatic hydrocarbons. the bioluminescence expressed by the luc gene in the reporter was well triggered by the promoter when it was exposed to 2,3-dihydroxybiphenyI (2,3-DHBP) at 0.5 to 1 mM concentrations. The bioluminescent response was more extensive when the reporter strain was exposed to 5 mM catechol and 2 mM 4-chlorocatechol. These different types of bioluminescent responses by E. coli SHP3 appeared to be characterized by the nature of the aromatics to stress. Since E. coli SHP3 responded to 2,3-DHBP quite sensitively, this reporter strain could be applied for detecting some catecholic pollutants.

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Pseudomonas sp. DJ77 균주에서 Extradiol Dioxygenase를 암호화하는 phnQ 유전자의 클로닝과 대장균에서의 발현 (Cloning of phnQ Gene Encoding Extradiol Dioxygenase from Pseudomonas sp. DJ77 and Its Expression in Escherichia coli)

  • 신희정;박용춘;민경희;김치경;임재윤;김영창
    • 미생물학회지
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    • 제33권1호
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    • pp.22-26
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    • 1997
  • Pseudomonas sp. DJ77의 게놈 library로부터 phenanthrene 분해에 관련된 유전자를 포함하는 약 5-kb의 XhoI 절편을 pBLUESCRIPT SK(+)로 클로닝하였으며, 이 재조합 plasmid를 pUPX5라 명명하였다. 이 재조합 균주에 catechol과 2,3-dihydroxybiphenyl 용액을 분무하면 노란색의 meta-cleavage 화합물이 생성됨을 관찰 할 수 있었다. 그리고 효소 활성을 측정한 결과 catechol에서보다 2,3-dihydroxybiphenyl에 더 큰 활성을 나타냈다. 이 부분에 존재하는 2,3-dihydroxybiphenyl 1,2-dioxygenase 유전자의 위치를 결정하고, 이 유전자를 phnQ라 명명하였다.

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디젤오염지역에서 분리한 세균 Sphingomonas sp. 3Y의 석유계 탄화수소분해특성 (Characterization of Petroleum Hydrocarbon Degradation by a Sphingomonas sp. 3Y Isolated from a Diesel-Contaminated Site.)

  • 안영희;정병길;성낙창;이영옥
    • 생명과학회지
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    • 제19권5호
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    • pp.659-663
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    • 2009
  • 장기간 경유로 오염된 지역의 토양으로부터 분리한 세균 3Y는 석유계 탄화수소를 구성하는 다양한 화합물을 유일 탄소원으로하여 성장하였다. Sphingomonas sp. 3Y는 지방족 화합물은 물론이고 방향족 화합물을 이용해서 성장할 수 있었다. 지방족 화합물로서는 hexane과 hexadecane을 이용하여 성장하였고, 한편 방향족 화합물로서는 BTEX는 물론이고 phenol, biphenyl, 또는 phenanthrene을 유일 탄소원으로 이용하여 성장하였다. 본 균주는 indole과 catechol을 이용한 실험결과 방향족 탄화수소의 생분해 과정에서 맨 첫 단계 반응에 관여하는 효소인 aromatic ring dioxygenase 활성과 benzene 환을 깨는 효소인 meta-cleavage dioxygenase 활성을 나타내었다. Sphingomonas sp. 3Y의 16S rRNA 유전자의 염기서열 분석과 계통수 작성 결과 본 균주는 ${\alpha}$-Proteobacteria인 Sphingomonas속에 해당하였으며 지금까지 잘 알려진 석유계 탄화수소를 분해하는 Sphingomonas sp. 균주들과는 다른 cluster를 형성하였다. 다양한 석유계 탄화수소 성분을 이용하여 성장하는 Sphingomonas sp. 3Y는 유류로 오염된 토양의 복원에 유용하게 사용될 것으로 여겨지며 이 균주의 최적 분해 조건을 조사한다면 그 결과는 이 균주가 분리된 오염지역의 생물학적 분해를 최적화하는데 기여할 것이다.

Extradiol Cleavage of Two-ring Structures of Biphenyl and Indole Oxidation by Biphenyl Dioxygenase in Commamonas Acidovorans

  • On, Hwa-Young;Lee, Na-Ri;Kim, Young-Chang;Kim, Chi-Kyung;Kim, Young-Soo;Park, Yong-Keun;Ka, Jong-Ok;Lee, Ki-Sung;Min, Kyung-Hee
    • Journal of Microbiology and Biotechnology
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    • 제8권3호
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    • pp.264-269
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    • 1998
  • Commamonas acidovorans SMN4 showed wide growth substrate spectra for various aromatic hydrocarbons. Strain SMN4 was able to grow on biphenyl producing a meta-cleavage compound, yellow 2-hydroxy-6-oxophenylhexa-2,4-dienoic acid with a spray of 2,3-dihydroxybiphenyl, while it also grew on catechol, developing yellow 2- hydroxymucoic semialdehyde with a spray of 100 mM catechol. Thus these results indicate that two-ring structures of biphenyl were cleaved by meta-mode in upper and lower pathways. Strain SMN4 metabolized various substituted biphenyl compounds and xylene to the corresponding benzoate derivatives through oxidation of the ring structures. It was clearly shown that biphenyl can be a common inducer in the oxidation of biphenyl and 2,3-dihydroxybiphenyl. Various compounds were examined for their suitability to serve as substrates for indole oxidation, indicating that biphenyl, benzoate, and succinate are quite good inducers of indigo production due to the activity of biphenyl dioxygenase. This results suggest that indigo formation is by means of the combined activities of biphenyl dioxygenase and tryptophanase.

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Isolation and Characterization of a Rhodococcus Species Strain Able to Grow on ortho- and para-Xylene

  • Jang Jung Yeon;Kim Dockyu;Bae Hyun Won;Choi Ki Young;Chae Jong-Chan;Zylstra Gerben J.;Kim Young Min;Kim Eungbin
    • Journal of Microbiology
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    • 제43권4호
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    • pp.325-330
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    • 2005
  • Rhodococcus sp. strain YU6 was isolated from soil for the ability to grow on o-xylene as the sole carbon and energy source. Unlike most other o-xylene-degrading bacteria, YU6 is able to grow on p-xylene. Numerous growth substrate range experiments, in addition to the ring-cleavage enzyme assay data, suggest that YU6 initially metabolizes 0- and p-xylene by direct aromatic ring oxidation. This leads to the formation of dimethylcatechols, which was further degraded largely through meta-cleavage path-way. The gene encoding meta-cleavage dioxygenase enzyme was PCR cloned from genomic YU6 DNA using previously known gene sequence data from the o-xylene-degrading Rhodococcus sp. strain DK17. Subsequent sequencing of the 918-bp PCR product revealed a $98\%$ identity to the gene, encoding meth-ylcatechol 2,3-dioxygenase from DK17. PFGE analysis followed by Southern hybridization with the catechol 2,3-dioxygenase gene demonstrated that the gene is located on an approximately 560-kb megaplasmid, designated pJY J1

Sequence Characteristics of xylJQK Genes Responsible for Catechol Degradation in Benzoate-Catabolizing Pseudomonas sp. S-47

  • Park, Dong-Woo;Lee, Jun-Hun;Lee, Dong-Hun;Lee, Kyoung;Kim, Chi-Kyung
    • Journal of Microbiology and Biotechnology
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    • 제13권5호
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    • pp.700-705
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    • 2003
  • Pseudomonas sp. S-47 is capable of degrading benzoate and 4-chlorobenzoate as well as catechol and 4-chlorocatechol via the meta-cleavage pathway. The three enzymes of 2-oxopenta-4-enoate hydratase (OEH), acetaldehyde dehydrogenase (acylating) (ADA), and 2-oxo-4-hydroxypentonate aldolase (HOA) encoded by xylJQK genes are responsible for the three steps after the meta-cleavage of catechol. The nucleotide sequence of the xylJQK genes located in the chromosomal DNA was cloned and analyzed. GC content of xylJ, xylQ, and xylK was 65% and consisted of 786, 924, and 1,041 nucleotides, respectively. The deduced amino acid sequences of xylJ, xylQ, and xylK genes from Pseudomonas sp. S-47 showed 93%, 99%, and 99% identity, compared with those of nahT, nahH, and nahI in Pseudomonas stutzeri An10. However, there were only about 53% to 85% identity with xylJQK of Pseudomonas putida mt-2, dmpEFG of P. putida CF600, aphEFG of Comamonas testosteroni TA441, and ipbEGF of P. putida RE204. On the other hand, the xylLTEGF genes located upstream of xylJQK in the strain S-47 showed high homology with those of TOL plasmid from Pseudomonas putida mt-2. These findings suggested that the xylLTEGFIJQK of Pseudomonas sp. S-47 responsible for complete degradation of benzoate and then catechol via the meta-pathway were phylogenetically recombinated from the genes of Pseudomonas putida mt-2 and Pseudomonas stutzeri An10.

페놀 분해 Rhodococcus sp. DGUM 2011의 분리 및 특성

  • 오정석;한영환
    • 한국미생물·생명공학회지
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    • 제25권5호
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    • pp.459-463
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    • 1997
  • A bacterium DGUM 2011 has been selected from various samples of industrial wastewater and soil. Based on the morphological and physiological characteristics, the isolate DGUM 2011 was identified as Rhodococcus sp. and named as Rhodococcus sp. DGUM 2011. The optimal temperature and pH for the cell growth of Rhodococcus sp. DGUM 2011 were 37$\circ$C and 7.6, respectively. When phenol was added to the minimal media as a sole source of carbon and energy, the concentrations of maximum and optimum for cell growth was 0.10% and 0.08%, respectively. When 0.05% phenol was given in the minimal media, Rhodococcus sp. DGUM 2011 completely utilize it within 24 hrs. The isolate could utilize benzoic acid, p-hydroxybenzoate, p-cresol, tyrosine and phloroglucinol. The isolate possessed both catechol 1,2-dioxygenase and 2,3-dioxygenase activity. However, the activity of catechol 1,2-dioxygenase was much higher than that of 2,3-dioxygenase, which suggests that the isolate might degrade phenol via both ortho- and meta-cleavage, mainly via ortho-cleavage.

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정면적법을 이용한 불연속면의 특성화 및 사면안정해석: 충북 보은군 내북면 창리 지역 (Analysis of Slope Stability and Property of Discontinuities Using Square-Inventory Method: The Changri area, Boeun-Gun, Chungbuk)

  • 최병렬;정상원
    • 터널과지하공간
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    • 제18권1호
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    • pp.20-32
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    • 2008
  • 정면적법을 이용하여 연구지역의 사면 안정성 분석을 하였으며, 이 방법은 선조사 방법이나 면적조사법을 이용하는 것보다 빠르게 유추할 수 있는 방법이다. 연구지역은 충북 보은군 창리 일대에 속해 있는 옥천누층군의 지층으로 구성되어 있으며 연구지역을 포함하여 천매암으로 구성된 지층에서 여러 종류의 사면 파괴 유형이 발생했음을 관찰하였다. 이러한 변성퇴적암의 물리적 특성은 광물입자가 극세립질의 석영과 견운모로 구성되었으며 층리(또는 편리) 구조와 파랑벽개구조를 따라 층리에 평행하게 배열되어 암석의 공학적 특성에 영향을 끼치는 것으로 분석되었다. 연구지역 내 3개 지점을 선정하여 각각 사면의 특성을 대표할 수 있는 $1m{\times}1m$ 사면을 2 또는 3 개소를 선택하여 사면 안정성분석을 시도하였다. 분석결과 정면적법을 이용한 사면의 안정성분석에서 3개 연구지점 모두에서 전도파괴와 쐐기파괴의 가능성이 매우 높은 것으로 분석되었으며 또한 연구지점 2의 경우 평면파괴의 가능성은 미약한 것으로 분석되었으나 실제 현장의 노두에서는 평면파괴가 발생한 곳이 다수 관찰되었다. 변성퇴적암에서는 화성암과는 다른 불연속면 구조인 층리(또는 편리)와 벽개구조가 함께 존재하기 때문에 사면 안정성 분석에서 절리구조와 더불어 반드시 고려해야 할 중요한 요소이다.