• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.386-390
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• 2006

A green fluorescent protein-based Pseudomonas putida reporter was successfully constructed and shown to be capable of detecting oxidative stress. In this whole-cell reporter, the promoter of the paraquat-inducible ferredoxin-$NADP^+$ reductase (fpr) was fused to a promoterless gfp gene on a broad-host-range promoter probe vector. Pseudomonas putida KT2440 harboring this reporter plasmid exhibited an increased level of gfp expression in the presence of redox-cycling agents (paraquat and menadione), hydrogen peroxide, and potential environmental pollutant chemicals such as toluene, paint thinner, gasoline, and diesel. Induction of fpr in the presence of these chemicals was confirmed using Northern blot analysis.

• Journal of Microbiology
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• 제44권6호
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• pp.671-673
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• 2006

A new 4.87 kb Escherichia-Pseudomonas shuttle vector has been constructed by inserting a 1.27 kb DNA fragment with a replication origin of a Pseudomonas plasmid pRO1614 into the 3.6 kb E. coli plasmid pBGS18. This vector, designated pJH1, contains an aminogly-coside phosphotransferase gene (aph) from Tn903, a lacZ' gene for $\alpha$-complementation and a versatile multiple cloning site possessing unique restriction sites for EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, BspMI, PstI, SphI, and HindIII. When pJH1 was transformed into E. coli DHS${\alpha}$ and into P. putida HK-6, it was episomally and stably maintained in both strains. In addition, the enhanced green fluorescent protein (EGFP) gene which was transcriptionally cloned into pJH1 rendered E. coli cells fluorescence when its transformants were illuminated at 488 nm.

• Journal of Microbiology and Biotechnology
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• 제32권8호
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• pp.989-1002
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• 2022

Cephalopods, in particular octopus (Octopus vulgaris), have the ability to alter their appearance or body pattern by showing a wide range of camouflage by virtue of their chromatophores, which contain nanostructured granules of ommochrome pigments. Recently, the antioxidant and antimicrobial activities of ommochromes have become of great interest; therefore, in this study, the pH-dependent redox effect of the extraction solvent on the antioxidant potential and the structural characterization of the pigments were evaluated. Cell viability was determined by the microdilution method in broth by turbidity, MTT, resazurin, as well as fluorescence microscopy kit assays. A Live/Dead Double Staining Kit and an ROS Kit were used to elucidate the possible inhibitory mechanisms of ommochromes against bacterial and fungal strains. The results obtained revealed that the redox state alters the color changes of the ommochromes and is dependent on the pH in the extraction solvent. Natural phenoxazinone (ommochromes) is moderately toxic to the pathogens Staphylococcus aureus, Bacillus subtilis, Salmonella Typhimurium and Candida albicans, while the species Pseudomonas aeruginosa and Pseudomonas fluorescens, and the filamentous fungi Aspergillus parasiticus, Alternaria spp. and Fusarium verticillioides, were tolerant to these pigments. UV/visible spectral scanning and Fourier- transform infrared spectroscopy (FTIR) suggest the presence of reduced ommatin in methanol/ HCl extract with high intrinsic fluorescence.

• 식물병연구
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• 제24권3호
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• pp.242-247
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• 2018

Pseudomonas chlororaphis O6 induces systemic tolerance in plants against drought stress. A volatile, 2R, 3R-butanediol, produced by the bacterium causes partial stomatal closure, thus, limiting water loss from the plant. In this study, we report that applications of extracellular polymeric substances (EPS) from P. chlororaphis O6 to epidermal peels of leaves of Arabidopsis thaliana also reduce the size of stomatal openings. Growth of A. thaliana seedlings with applications of the EPS from P. chlororaphis O6 reduced the extent of wilting when water was withheld from the plants. Fluorescence measurements showed photosystem II was protected in the A. thaliana leaves in the water stressed EPS-exposed plants. These findings indicate that P. chlororaphis O6 has redundancy in traits associated with induction of mechanisms to limit water stress in plants.

• Journal of Microbiology and Biotechnology
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• 제8권2호
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• pp.111-118
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• 1998

A bacterium producing unique antistaphylococcal substance (ASS) was isolated from soil samples. The isolated strain KUH-001 was identified to belong to Pseudomonas species from the characteristic properties of its fluorescence and cellular 3-hydroxy fatty acid composition, etc. The ASS component was purified by procedures employing activated carbon adsorption, column chromatography with silica gel, preparative TLC and HPLC. This compound could also be purified mainly by repeating of trituration and precipitation with chilled ether. Purified ASS with a m.p. value of $140~142^{\circ}C$ showed marked stability at high temperature (at $121^{\circ}C$ for 10 min) and extreme pHs (in 1N HC1 and 1N NaOH for 1 day) without significant loss of antibiotic activity. From spectral data of UV, IR, NMR, and FAB-MS, the compound was elucidated as 2-heptyl-4-hydroxyquinoline N-oxide (HHQO). Under the conditions employed, HHQO exhibited a narrow antimicrobial spectrum. active particularly against Staphylococcus aureus including the methicillin resistant strain. Moreover, it did not induce resistance, and besides, interacted synergistically with certain antibiotics such as vancomycin or erythromycin.

• 한국지하수토양환경학회지:지하수토양환경
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• 제12권2호
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• pp.1-8
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• 2007

토양 및 지하수의 오염 확산을 방지하면서 오염물을 처리할 수 있는 생물벽체(Biobarrier)에 적용하기 위해 회분식실험과 칼럼 실험을 실시하였다. 회분식 실험에서는 순수 호기성 미생물 Pseudomonas fluorescens의 글루코즈와 당밀에 대한 동역학 상수를 산정하였다. P. fluorescens의 최대비성장속도(Vmax)는 글루코즈와 당밀을 사용하였을 때 $23^{\circ}C$에서 각각 $0.246\;hr^{-1}$, $0.073\;hr{-1}$로 글루코즈의 비성장속도가 빠르지만, P. fluorescence 성장 시 당밀에 대한 지체기가 없으며, 경제성을 고려하여 당밀을 칼럼실험의 탄소원으로 사용하였다. 일정수두 칼럼실험에서 P. fluorescence를 접종하고 영양원을 주입하면서 칼럼 내 토양의 투수계수 감소를 측정한 결과 영양원 주입 6일 만에 투수계수는 $4.1{\times}10^{-2}\;cm/sec$에서 $2.8{\tims}10^{-4}\;cm/sec$로 $6.8{\times}10^{-3}$배 감소하였고, 미생물량은 유입부에서 최대값을 나타냈다. 그러나 투수계수의 감소를 야기하는 미생물의 성장은 탄소원이 아닌 전자수용체(용존산소)의 농도에 의하여 제한되었다.

• 대한환경공학회지
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• 제35권6호
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• pp.449-455
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• 2013

소규모 하수처리시스템에 다양한 반송비와 고정식 담체를 적용하여 합성폐수에서 질소, 인의 제거를 검토하였다. 담체를 첨가 시 질소의 제거율은 40.1%에서 65.1%로 증가하였으나 COD와 인의 제거에는 영향이 없었다(<5%). 반송을 통해 유량을 2배(2Q)로 증가시킨 경우 질소의 제거율은 7% 정도가 증가하였으나, 인 제거율은 반송비가 증가됨에 따라 31.8에서 26.6%로 감소하였다. 무산소조에서 담체에 부착된 미생물은 약 73.4 $mg/cm^2$로 존재하였으며, FISH분석 결과 탈질 미생물인 Pseudomonas aeruginosa 존재 (약 58%)를 확인하였다.

• 한국식품영양과학회지
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• 제30권4호
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• pp.630-634
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• 2001

가공유는 우유에 과즙 등을 첨가하여 우유의 성분을 변화시킨 우유로서 백색 시유와는 달리 가공공정 중 각종 첨가물에 의해 미생물이 오염되어 품질악화를 초래할 수도 있다. 미생물 오염에 의한 가공유의 품질악화를 방지하고 품질개선을 위해 chitosan의 이용 가능성을 검토하였다. 시판 가공유에서 api 20E kit 와 api 20C AUX kit 로 동정한 결과 Saccharomyces cerevisiae와 Pseudomonas fluorescence를 분리동정하였다. 0.01%, 0.03%, 0.05%, 0.1% chitosan 을 함유한 YM broth에서 분리 yeast의 성장을 측정한 결과 0.03% chitosan 첨가구에서 배양 12시간후 성장이 완전히 저해되었다. 분리 P. fluorescence는 chitosan 0.03%를 함유한 TSB 에서 2~3 l$og_{10}$ cycle 성장이 억제되었다. Chitosan 0.03%를 첨가한 가공 시유를 제조하여 15일 동안 $4^{\circ}C$, 1$0^{\circ}C$, $25^{\circ}C$에서 저장하면서 품질특성을 조사한 결과 $4^{\circ}C$와 1$0^{\circ}C$에서는 대조구와 뚜렷한 차이를 나타내지 않았으나 $25^{\circ}C$ 저장의 경우 대조구는 저장 2일째부터 이화학적 변화(pH, 산도)를 나타낸 반면 chitosan 첨가구의경우 저장 15일 동안 저장초기와 거의 같은 수준을 나타내었다. Chitosan 첨가 가공유는 5% 유의수준에서 taste는 감소하였으며 texture는 증가하여, chitosan 첨가에 의한 가공유의 기호성 저하는 없었다.

• The Plant Pathology Journal
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• 제25권1호
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• pp.77-85
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• 2009

Pseudomonas fluorescens strain Gpf01, isolated from ginseng rhizosphere showed antiviral activity against Cucumber mosaic virus, when tested in a local host of CMV, Chenopodium amaranticolor. Transposon mutant library of Gpf01 was prepared using pGS9::Tn5 and the mutant Gpf01-RS19 was found to loose antiviral production. We developed primers from the flanking region of Tn5 and found a cosmid clone pAV1123, harboring 1.2 kb antiviral compound producing (avcf01) locus. When a sub-clone pPH9, which carried 9.3 kb region of pAV1123, was introduced into antivirus deficient P. fluorescens wild type strain B16, it exhibited antiviral activity. Using Tn3-gus mutagenesis and complementation analysis, it was found that the genes related to antiviral activity production resided in a 9.3 kb HindIII-HindIII fragment of pAV1123, indicating that the plasmid carries an essential genes promoting antiviral activity.

• Mycobiology
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• 제33권3호
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• pp.131-136
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• 2005

Infection structures were observed at the penetration sites on the leaves of cucumber plants inoculated with Colletotrichum orbiculare using a fluorescence microscope. The cucumber plants were previously drenched with suspension of bacterial strains Pseudomonas putida or Micrococcus luteus. The plants pre-inoculated with both bacterial strains were resistant against anthracnose after inoculation with C. orbiculare. To investigate the resistance mechanism by both bacterial strains, the surface of infected leaves was observed at the different time after challenge inoculation. At 3 days after inoculation there were no differences in the germination and appressorium formation of conidia of C. orbiculare as well as in the callose formation of the plants between both bacteria pre-inoculated and non-treated. At 5 days, the germination and appressorium formation of the fungal conidia were, however, significantly decreased on the leaves of plants pre-inoculated with M. luteus at the concentration with $1.0{\times}10^7\;cfu/ml$. Furthermore, callose formation of plants cells at the penetration sites was apparently increased. In contrast, there were no defense reactions of the plants at the concentration with $1.0{\times}10^6\;cfu/ml$ of M. luteus. Similarly, inoculation P. putida caused no plant resistance at the low concentration, whereas increase of callose formation was observed at the higher concentration. The results of this study suggest that the resistant mechanisms might be differently expressed by the concentration of pre-treatment with bacterial suspension.