• Journal of Microbiology and Biotechnology
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• 제15권6호
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• pp.1170-1177
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• 2005

Terminal-restriction fragment length polymorphism (T-RFLP) analysis was used to monitor inoculated oil-degrading microorganisms during bioremedial treatability tests. A pair of universal primers, fluorescently labeled 521F and 1392R, was employed to amplify small subunit rDNA in order to simultaneously detect two bacterial strains, Corynebacterium sp. IC10 and Sphingomonas sp. KH3-2, and a yeast strain, Yarrowia lipolytica 180. Digestion of the 5'-end fluorescence/labeled PCR products with HhaI produced specific terminal-restriction fragments (T-RFs) of 185 and 442 bases, corresponding to Corynebacterium sp. IC10 and Y. lipolytica 180, respectively. The enzyme NruI produced a specific T-RF of 338 bases for Sphingomonas sp. KH3-2. The detection limit for oildegrading microorganisms that were inoculated into natural environments was determined to be $0.01\%$ of the total microbial count, regardless of the background environment. When three oil-degrading microorganisms were released into oil-contaminated sand microenvironments, strains IC10 and 180 survived for 35 days after inoculation, whereas strain KH3-2 was detected at 8 days, but not at 35 days. This result implies that T-RFLP could be a useful tool for monitoring the survival and relative abundance of specific microbial strains inoculated into contaminated environments.

• 한국환경보건학회지
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• 제33권5호
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• pp.428-433
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• 2007

To clarify the pioneer group in the development of biofilms in high chlorine residual water, a semi-pilot model system was operated and 16S rDNA V3 targeted PCR-DGGE was submitted. Biofilm formation occurred rapidly in the model of a drinking water distribution system. It reached $10^3\;CFU/cm^2$ or more on the surface of stainless steel, PVC, and galvanized iron in chlorinated (1.0 mg/l) water within a week. Within a week, uncultured Proteobacteria- and Bacillales group-like sequences were detected and Sphingomonas-like sequences were identified from all season and all pipe materials tested. Hence Sphingomonas species were regarded as the potential pioneer group in the development of biofilm in drinking water and this results would be useful for the prevention of biofilm formation and safety of drinking tap water.

• KSBB Journal
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• 제21권6호
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• pp.428-432
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• 2006

본 연구는 미생물이 오염물 분해를 위해 황산염이 필요한 동전기 생물학적복원에서 황산염원의 종류가 제거효율에 미치는 영향을 조사하였다. 대표오염물과 적용 미생물은 각각 phenanthrene과 Sphingomonas sp. 3Y였다. Magnesium sulfate를 이용했을 때 Mg이온은 음극에서 전기적으로 생성되는 수산화이온과 결합하여 전해질 pH를 크게 감소시키고 미생물활성을 저해하는 현상이 관찰되었다. 따라서 ammonium sulfate와 disodium sulfate를 이용했을 때 전해질 및 토양 pH가 중성영역으로 유지되었으며 미생물 활성도 높게 유지되었다. 하지만 전자의 경우 ammonium이 미생물의 오염물 분해를 지연시켜 제거효율이 오히려 magnesium sulfate의 21.8%보다 12.0%로 더 낮게 나타났으며, 반면 후자의 경우 27.2%로 제거효율이 증가된 것을 관찰할 수 있었다. 이와 같은 제거효율의 차이는 장기간의 처리에서는 더욱 두드러질 것으로 예상된다.

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 2004년도 임시총회 및 추계학술발표회
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• pp.181-185
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• 2004

Electrokinetic bioremediation was conducted on phenanthrene-contaminated soil to study the effects of soil temperature and pH on microbial population and removal efficiency at different current densities from 0.63 to 3.13 mA cm$^{-2}$ . Microorganism used in the biodegradation of phenanthrene was Sphingomonas sp. 3Y, which was isolated from a diesel-contaminated site. The microorganism was successfully penetrated into the contaminated soil by electrokinetic phenomena and the highest microbial population was observed in the middle region of soil specimen where soil pH was near neutral. Therefore, phenanthrene removal occurred mainly at anode and middle parts of soil specimen due to a relatively high microbial population. Also, the highest removal efficiency of 68.8% was obtained at 1.88 mA cm$^{-2}$ while low degradation was detected at 3.13 mA cm$^{-2}$ . It was presumably because the soil temperature at 1.88 mAcm$^{-2}$ was close to the appropriate temperature of about 30'c while the temperature increase to above 45$^{\circ}C$ at 3.13 mA cm$^{-2}$ inhibited the microbial activity severely.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1998년도 한국생물과학협회 학술발표대회
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• pp.173.1-173
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• 1998

No Abstract, See Full Text

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 2004년도 임시총회 및 추계학술발표회
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• pp.186-190
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• 2004

The electrokinetic bioremediation employing electrolyte circulation method was carried out for the cleanup of phenanthrene-contaminated kaolinite, and microorganism used in the biodegradation of phenanthrene was Sphingomonas sp. 3Y. The electrolyte circulation method supplied ionic nutrientsand the microorganism into soil, and inhibited the significant pH change of soil by increasing the soil buffering capacity by providing phosphate buffer compounds. When the remediation process was conducted without surfactant, the removal efficiency of phenanthrene, at the initial concentration of 200 ppm, was 69% for only 7 days. Higher microbial population and lower phenanthrene concentration were observed in the anode and middle regions of soil specimen than in the cathode region. The higher density of microorganism was because the microbial movement was in the direction of the anode part due to the negative surface charge. When Triton X-100 and APG of 20 g/1 were used to improve the bioavailability of phenanthrene strongly adsorbed onto soil surface, about 90 and 39% of phenanthrene removal were obtained. Consequently, it was confirmed that the microorganism preferred APC to phenanthrene as carbon source and so the removal efficiency with APG decreased less than that without APG.

• Journal of Microbiology and Biotechnology
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• 제22권11호
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• pp.1532-1539
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• 2012

The ${\alpha}$-galactosidase-coding gene agaAJB13 was cloned from Sphingomonas sp. JB13 showing 16S rDNA (1,343 bp) identities of ${\leq}97.2%$ with other identified Sphingomonas strains. agaAJB13 (2,217 bp; 64.9% GC content) encodes a 738-residue polypeptide (AgaAJB13) with a calculated mass of 82.3 kDa. AgaAJB13 showed the highest identity of 61.4% with the putative glycosyl hydrolase family 36 ${\alpha}$-galactosidase from Granulicella mallensis MP5ACTX8 (EFI56085). AgaAJB13 also showed <37% identities with reported protease-resistant or Sphingomonas ${\alpha}$-galactosidases. A sequence analysis revealed different catalytic motifs between reported Sphingomonas ${\alpha}$-galactosidases (KXD and RXXXD) and AgaAJB13 (KWD and SDXXDXXXR). Recombinant AgaAJB13 (rAgaAJB13) was expressed in Escherichia coli BL21 (DE3). The purified rAgaAJB13 was characterized using p-nitrophenyl-${\alpha}$-D-galactopyranoside as the substrate and showed an apparent optimum at pH 5.0 and $60^{\circ}C$ and strong resistance to trypsin and proteinase K digestion. Compared with reported proteaseresistant ${\alpha}$-galactosidases showing thermolability at $50^{\circ}C$ or $60^{\circ}C$ and specific activities of <71 U/mg with or without protease treatments, rAgaAJB13 exhibited a better thermal stability (half-life of >60 min at $60^{\circ}C$) and higher specific activities (225.0-256.5 U/mg). These sequence and enzymatic properties suggest AgaAJB13 is the first identified and characterized Sphingomonas ${\alpha}$-galactosidase, and shows novel protease resistance with a potential value for basic research and industrial applications.

• 생명과학회지
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• 제23권12호
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• pp.1420-1427
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• 2013

이전에 새로운 아세틸알긴산 아세틸분해효소(acetylalginate esterase, AcAlgE)를 Sphingomonas sp. MJ-3 균주로부터 클로닝하고 특성을 보고한 바 있다. 본 연구에서는 Pseudomonas aeruginosa로부터 얻은 아세틸알긴산을 분해하는데 미치는 MJ-3 AcAlgE와 KS-408 알긴산 분해효소의 상승효과를 고-자기장 $^1H$-NMR과 peptide column을 장착한 FPLC를 이용하여 조사하였다. 알긴산 분해효소 coupled assay 법으로 측정한 결과 AcAlgE 효소는 산가수 분해하여 얻은 저분자 아세틸알긴산을 분해하는 것보다 고분자 아세틸알긴산을 분해하는 경우 낮은 활성을 보였다. 아세틸알긴산을 알긴산 분해효소로 분해하는 경우 먼저 AcAlgE로 아세틸알긴산의 아세틸기를 분해하여 제거한 후에야 KS-408 알긴산 분해효소의 활성이 높은 것으로 나타났다. 이러한 결과는 재조합 AcAlgE는 알긴산 분해효소에 의한 아세틸알긴산의 분해효과를 상승시킨다는 사실을 보여준다.

• 미생물학회지
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• 제34권4호
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• pp.194-199
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• 1998

한강의 본류와 만나는 탄천과 중랑천에서 16S rDAN를 증폭하고 부분적인 염기서열 분석을 통하여 한강의 세균 다양성을 결정하였다. 총 27개의 클론을 분리하였으며 RFLP를 이용하여 7개의 group으로 나누었다. 탄천의 15개 클론은 4개의 group으로 나뉘어졌으며 가장 많은 클론을 포함하는 group(HT-1 클론)은 class Proteobacteria의 ${\delta}$-subdivision에 속하는 Acrobacter cryaerophilius와 높은 유사도를 보였으며, 다른 두 group(HT-6과 HT-9 클론)은 모두 clas Cytophagales에 속하였다. 중랑천의 12개의 클론은 3개의 group으로 나뉘어졌으며 가장 많은 클론을 보이는 group(HJ-1 클론)은 class Proteobacteria의 ${\alpha}$-subdivision에 속하는 Sphingomonas sp. 와 높은 유사도를 나타내었다. 전체적으로는 Proteobateria(alpha, beta and delta subdivision), Cytophagales와 Actinomycetales가 검출되었다.

• BMB Reports
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• 제32권4호
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• pp.399-404
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• 1999

2,3-Dihydroxybiphenyl 1,2-dioxygenase (2,3-DHBD), which catalyzes the ring meta-cleavage of 2,3-dihydroxybiphenyl, is encoded by the phnQ gene of biphenyl- and phenanthrene-degrading Pseudomonas sp. strain DJ77. We determined the nucleotide sequence of a DNA fragment of 1497 base pairs which included the phnQ gene. The fragment lncluded an open reading frame of 903 base pairs to accommodate the enzyme. The predicted amino acid sequence of the enzyme subunit consisted of 300 residues. In front of the gene, a sequence resembling an E. coli promoter was identified, which led to constitutive expression of the cloned gene in E. coli. The deduced amino acid sequence of the PhnQ enzyme exhibited 85.6% identity with that of the corresponding enzyme in Sphingomonas yanoikuyae Q1 (formerly S. paucimobilis Q1) and 22.1% identity with that of catechol 1,2,3-dioxygenase from the same DJ77 strain. PhnQ showed broader substrate preference than previously-cloned PhnE, catechol 2,3-dioxygenase. Ten amino acid residues, considered to be important for the role of extradiol dioxygenases, were conserved.