다환방향족 탄화수소에 오염된 토양에서 분리한 Pseudomonas sp. KK1의 생리학적 특성

윤병준,강형일,오덕철 제주대학교 생명과학연구소 2001 제주생명과학연구 Vol.4 No.-

Strain KK1 was isolated from soil contaminated with polycyclic aromatic hydrocarbons and able to mineralize anthracene, naphthalene, and phenanthrene. In this study, strain KK1 was tested to evaluate carbazole-degrading potential using radiorespirometry. When KK1 was pre-grown on phenanthrene the cells were able to mineralize carbazole much more rapidly, suggesting a possible close linkage between the pathways for catabolism of carbazole and phenanthrene. Such an increase in carbazole degradation was not appreciable when KK1 was pre-grown on naphthalene. Strain KK1 was identified as the genus Pseudomonas with over 90 % confidence based on BIOLOG system and FAME analysis. Analysis of PLFAs extracted from kk1 cells on carbazole medium revealed that lipids 10:0 3OH, 17:0 cyclo, and 18:0 were representatives produced or significantly increased in response to carbazole. Tests for antibiotics provided the data that strain KK1 was resistant to antibiotic ampicillin, but susceptibile to chloramphenicol, gentamycin, kanamycin, streptomycin, and tetracycline. Strain KK1 demonstrated strong resistance to most heavy metals such as Ba, Cd, Fe, Hg, Pb used in this study.

광합성 세균 Rhodospirillum rubrum S1이 생성한 Catalase-Peroxidase의 부분 정제 및 특성 규명

김영미,이동헌,강형일,오덕철 濟州大學校 基礎科學硏究所 2001 基礎科學硏究 Vol.14 No.2

호기적으로 배양한 광합성 세균 Rhodospirillum rubrum SI은 5 가지의 catalase를 생성함을 알 수 있었다. 그 중 peroxidase의 기능도 동시에 갖는 catalase-peroxidase(Cat-3)를 부분 정제하고 그 특성을 조사하였다. 부분정제된 catalase-peroxidase의 수율은 catalase가 1.6% 그리고 peroxidasse가 5.1% 였으며, 정제 배수가 4.6배와 14배로 증가한 효소를 얻을 수 있었는데, catalase보다 peroxidase의 정제배수가 더 높았다. pH에 대한 영향은 catalase활성은 pH6에서, peroxidase활성은 pH5에서 가장 높게 나타났다. 온도에 대한 영향은 catalase와 peroxidase 활성이 모두 30℃에서 최고의 활성을 보이는 것으로 나타났으며, 온도에 대한 효소의 안정성은 50℃에서 catalase가 peroxidase보다 더 안정함을 알 수 있었다. 부분정제된 catalase-peroxidase에 유기용매와 10mM 3-amino-1,2,4-triazole을 처리한 결과, 유기용매에 대해 catalase의 활성은 79%, peroxidase의 활성은 85%까지 억제됨을 알 수 있었으며, 10mM 3-amino-1,2,4-triazole에 대한 각 효소의 활성은 그대로 유지되고 있었다. 전형적인 heme단백질 효소들의 저해제로 알려진 NaCN, NaN3, NH20H를 농도별로 처리한 결과, NaCN인 경우 catalase는 8.72×10-s M의 농도에서, peroxidase는 5.1×10-s M의 농도에서, NaN3에서 catalase는 4.2×10-7 M, peroxidase는 3.2×10-7 M, NH20H에서는 catalase인 경우는 2.0×10-7 M, peroxidase인 경우는 2.5×10-7M 농도에서 억제되었으며, 이들 저해제들은 catalase활성과 peroxidase활성을 동시에 저해하는것으로 나타났다. Five different catalases have been already found in the aerobically grown photosynthetic bacterium Rhodospirillum rubrum S1. Among them, a bifunctional catalase-peroxidase, designated Cat-3 was partially purified and characterized. The enzyme was purified through four steps in 1.6% yield for catalatic activity and 5.1% yield for peroxidatic activity. On the basis of catalatic activity, the protein purification increased nearly 4.6-fold, whereas for peroxidatic activity, an approximately 14-fold purification was achieved. The optimum pHs of the catalatic and peroxidatic activities were 6 and 5, respectively, and the optimum temperature for both activities was 30℃. The catalatic activity of the enzyme maintained at 50℃ for lh was more stably than peroxidatic activity. The catalatic and peroxidatic activities were inhibited about 79 % and 85 % by exposure to organic solvent(ethanol/chloroform), respectively. Both enzymatic activities were not neally inhibited by lOmM 3-amino-1,2,4-triazole. Fifty percent enzyme inhibition of the catalatic activity was reached with 2.0×10^(-7)M, hydroxylamine, 4.2×10^(-7)M azide and 8.7×10^(-6)M cyanide, and that of the peroxidatic activity was obtained with 2.5×10^(-7)M hydroxylamine, 3.2×10^(-7)M azide and 5.1×10^(-6)M cyanide.

제주연안 갯녹음(백화) 지역의 해수에 분포하는 세균군의 분자생물학적 분석

강봉조,김미란,윤병준,이동헌,오덕철,강형일 濟州大學校 基礎科學硏究所 2002 基礎科學硏究 Vol.15 No.2

본 연구에서는 갯녹음(백화)현상이 세균생태계와 어떤 관련이 있는지에 대한 기초자료 및 정보를 얻기 위하여 갯녹음 현상이 일어난 제주도 성산과 강정지역 연안의 해수에 존재하는 세균군을 16S rRNA 증폭기법을 이용하여 조사하였다. 강정지역에서는 Alcanivorax, Paracoccus, Damselae, Pseudomonas, Rhodovulum, Silicibacter, Sulfitobacter, Roseobacter 등 다양한 종류의 세균이 분포되어 있었으며, Alcanivorax가 20%의 빈도로 가장 많이 나타났다. 반면, 성산 지역에서는 Pseudomonas속 균주가 우점종으로서 존재하였으며, Pseudomonas tolaasii(혹은 Pseudomonas corrugata)와 유연관계가 가까운 세균은 44%, Pseudomonas mandeli와 가까운 세균이 24%, Verrucomicrobiales와 가까운 세균은 4%, 기타 동정되지 않은 세균은 세 group으로 구분되었으며, 각각 8%, 8%, 12%를 차지하여, 두 곳에 분포되어 있는 세균군상이 상당한 차이점이 있음을 확인하였다. 갯녹음 지역인 강정과 성산 해수의 8월(표품 채집시기)의 수온은 27℃-27.5℃, 염분의 농도는 30.24-30.60%, pH는 8.23-8.36, 용존산소량(DO)은 각각 7.20-7.28로, 두 지역에서 매우 비슷한 것으로 조사된 바 있는데, 이는 수온이나 염분의 농도, 또는 pH보다는 다른 원인에 의하여 두 갯녹음 지역에 분포하는 세균군의 차이를 가져왔음을 제시해 주었다. In this study, the bacterial communities distributed in sea water of the whitening areas of Gangjeong and Seongsan, Jeju-do have been analyzed using the PCR amplification of 16S rRNA to obtain fundamental data and information on relationship of the whitening phenomenon and microbial ecosystem. In Gangjeong, diverse bacteria such as Alcanivorax, Paracoccus, Damselae, Pseudomonas, Rhodovulum, Silicibacier, Sulfitobacter, and Roseobacier have been found, and Alcanivorax was the most abundant clone. The most abundant clone from Seongsan was Pseudoinonas, of which Pseudomonastolaasii and Pseudonionas mandeli were most abundantly occurred in the frequency of approx 44% and 24%, respectively. Approx 4% of the bacterial clones closest to firruconiicrobiales and other unidentified clones were also found in Seongsan, suggesting there is a great discrepancy between bacterial communities from the whitening areas of Seongsan and Gangjeong. The mean tem- perature, chlorine concentration, pH, and dissolved oxygen (DO) of the sea water of Gangjeong and Seongsan in August of 2001 (sampling period) was 27℃-27.5℃, 30.24-30.60%, pH 8.23-8.36,7.20-7.28 ㎎/ℓ, suggesting other environmental factors except for the factors mentioned above might result in difference of bacterial communities distributed in both areas.

Isoaltion and Characterization of Aniline Degrading Bacteria

Hyung Yeel Kahng,Mi Jeong Woo,Yong Keun Park 한국미생물생명공학회(구 한국산업미생물학회) 1992 한국미생물생명공학회 학술발표초록집 Vol.1992 No.1

Molecular Detection of Catabolic Genes for Polycyclic Aromatic Hydrocarbons in the Reed Rhizosphere of Sunchon Bay

Hyung-Yeel Kahng,Kye-Heon Oh 한국미생물학회 2005 The journal of microbiology Vol.43 No.6

This study focused on detecting catabolic genes for polycyclic aromatic hydrocarbons (PAHs) distributed in the reed rhizosphere of Sunchon Bay, Korea. These marsh and mud environments were severely affected by human activities, including agriculture and fisheries. Our previous study on microbial roles in natural decontamination displayed the possibility that PAH-degrading bacteria, such as Achromobacter sp., Alcaligenes sp., Burkholderia sp. and Pseudomonas sp. play an important decontamination role in a reed rhizosphere. In order to gain further fundamental knowledge on the natural decontamination process, catabolic genes for PAH metabolism were investigated through PCR amplification of dioxygenase genes using soil genomic DNA and sequencing. Comparative analysis of predicted amino acid sequences from 50 randomly selected dioxygenase clones capable of hydroxylating inactivated aromatic nuclei indicated that these were divided into three groups, two of which might be originated from PAH-degrading bacteria. Amino acid sequences of each dioxygenase clone were a part of the genes encoding enzymes for initial catabolism of naphthalene, phenanthrene, or pyrene that might be originated from bacteria in the reed rhizosphere of Sunchon Bay.

Molecular Characteristics of Pseudomonas rhodesiae Strain KK1 in Response to Phenanthrene

( Hyung Yeel Kahng ) 한국미생물생명공학회 2002 Journal of microbiology and biotechnology Vol.12 No.5

Characterization of strain HY99, a novel microorganism capable of aerobic and anaerobic degradation of aniline

Kahng, Hyung-Yeel,Kukor, Jerome J.,Oh, Kye-Heon 濟州大學校 基礎科學硏究所 2001 基礎科學硏究 Vol.14 No.2

We have characterized a novel microorganism, strain HY99, which is capable of aerobic and anaerobic degradation of aniline. Strain HY99 was found to aerobically metabolize aniline via catechol and 2-hydroxymuconicsemialdehyde intermediates, and to transform aniline .via p-aminobenzoate in anaerobic environments. Physiological and biochemical tests revealed that strain HY99 was most similar to Delftia acidovorans, but unlike D. acidovorans, strain HY99 was able to metabolize aniline under anaerobic conditions linked with nitrate reduction. Phylogenetic analysis based on 16S rDNA sequencing also revealed that strain HY99 was closely related to D. acidovorans, with 96% overall similarity.

도서벽지(島嶼僻地) 유아(幼兒)의 사회화과정(社會化過程) : 언어(言語) 및 인지발달(認知發達)을 중심(中心)으로

고형일(Hyung Yeel Koh) 전남대학교 교육문제연구소 1981 교육연구 Vol.7 No.-

교육과정(教育課程)에 대한 신(新)·구교육사회학(舊教育社會學)의 리론적(理論的) 쟁점(爭点)

高炯一(Hyung-yeel Koh) 전남대학교 교육문제연구소 1980 교육연구 Vol.6 No.-

Molecular Detection of Catabolic Genes for Polycyclic Aromatic Hydrocarbons in the Reed Rhizosphere of Sunchon Bay

Kahng Hyung-Yeel,Oh Kye-Heon The Microbiological Society of Korea 2005 The journal of microbiology Vol.43 No.6

This study focused on detecting catabolic genes for polycyclic aromatic hydrocarbons (PAHs) distributed in the reed rhizosphere of Sunchon Bay, Korea. These marsh and mud environments were severely affected by human activities, including agriculture and fisheries. Our previous study on microbial roles in natural decontamination displayed the possibility that PAH-degrading bacteria, such as Achromobacter sp., Alcaligenes sp., Burkholderia sp. and Pseudomonas sp. play an important decontamination role in a reed rhizosphere. In order to gain further fundamental knowledge on the natural decontamination process, catabolic genes for PAH metabolism were investigated through PCR amplification of dioxygenase genes using soil genomic DNA and sequencing. Comparative analysis of predicted amino acid sequences from 50 randomly selected dioxygenase clones capable of hydroxylating inactivated aromatic nuclei indicated that these were divided into three groups, two of which might be originated from PAH-degrading bacteria. Amino acid sequences of each dioxygenase clone were a part of the genes encoding enzymes for initial catabolism of naphthalene, phenanthrene, or pyrene that might be originated from bacteria in the reed rhizosphere of Sunchon Bay.