• Title/Summary/Keyword: P. putida

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Aerobic Degradation of Tetrachloroethylene(PCE) by Pseudomonas stutzeri OX1

  • Ryoo, Doohyun;Shim, Hojae;Barbieri, Paola;Wood, Thomas K.
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2000.11a
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    • pp.207-208
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    • 2000
  • Since trichloroethylene (TCE), dichloroethylene (DCE), and vinyl chloride (VC) arise from anaerobic degradation of tetrachloroethylene (PCE) and TCE, there is interest in creating aerobic remediation systems that avoid the highly toxic VC and cis-DCE which predonominate in anaerobic degradation. However, it seemed TCE could not be degraded aerobically without an inducing compound (which also competitively inhibits TCE degradation). It has been shown that TCE induces expression of both the toluene dioxygenase of p. putida F1 as well as toluene-p-monooxygenase of P.mendocina KRI. We investigated here the ability of PCE, TCE, and chlorinated phenols to induce toluene-o-xylene monooxygenase (ToMO) from P.stutzeri OX1. ToMO has a relaxed regio-specificity since it hydroxylates toluene in the ortho, meta, and para positions; it also has a broad substrate range as it oxidizes o-xylene, m-xylene, p-xylene, toluene, benzene, ethylbenzene, styrene, and naphthalene; chlorinated compounds including TCE, 1, 1-DCE, cis-DCE, trans-DCE, VC, and chloroform : as well as mixtures of chlorinated aliphatics (Pseudomonas 1999 Maui Meeting). ToMO is a multicomponent enzyme with greatest similarity to the aromatic monooxygenases of Burkholderia pickettii PKO1 and P.mendocina KR1. Using P.sturzeri OX1, it was found that PCE induces P.mendocina KR1 Using P.situtzeri OX1, it was found that PCE induces ToMO activity measured as naphthalene oxygenase activity 2.5-fold, TCE induces 2.3-fold, and toluene induces 3.0 fold. With the mutant P.stutzeri M1 which does not express ToMO, it was also found there was no naphthalene oxygenate activity induced by PCE and TCE; hence, PCE and TCE induce the tow path. Using P.putida PaW340(pPP4062, pFP3028) which has the tow promoter fused to the reporter catechol-2, 3-dioxygenase and the regulator gene touR, it was determined that the tow promoter was induced 5.7-, 7.1-, and 5.2-fold for 2-, 3-, 4-chlorophenol, respectively (cf. 8.9-fold induction with o-cresol) : however, TCE and PCE did not directly induce the tou path. Gas chromatography and chloride ion analysis also showed that TCE induced ToMO expression in P.stutzeri OX1 and was degraded and mineralized. This is the first report of significant PCE induction of any enzyme as well as the first report of chlorinated compound induction of the tou operon. The results indicate TCE and chlorinated phenols can be degraded by P.stutzeri OX1 without a separate inducer of the tou pathway and without competitive inhibition.

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Antibacterial and Antifungal Effect by Artemisia lavandulaefolia Essential Oil (참쑥 정유의 항세균 및 항진균 효과)

  • 한규용
    • The Korean Journal of Food And Nutrition
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    • v.12 no.6
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    • pp.559-563
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    • 1999
  • Essential oil of Artemisia lavandulaefolia the chrysanthemum family plant used in the chinese medicine was extracted and antibacterial and antifungal activity with many kinds of the pathogenic bacterium and fungi was experimented by it. Ataphylococcus epidermidis and Streptococcus aureus gram positive bacterium at the concentration of 200ppm and Streptococcus mutans at the concentration of 1,000ppm showed the growth injibition effect of the cell. These showed statistically significant difference(p<0.05) Zymomonas mobilis Entrecoccus faecalis gram negative bacterium at the concentration of 200ppm and Pseudomonas putida at the concentration of 400ppm showedd the growth inhibition effect of the cell)p<0.05) V. Parahaemolyticus at the concentration of 800ppm and Pseudomonas aeruginosa at the concentration of 1,000ppm showed the growth inhibition effect of the cell(p<0.05) Candida albicans and Cryptococcus neoformans yeast-type fungi showed the gorwth inhibition effect of the cell at the concentration of 200ppm(p<0.05) Altenaria mali Aspergillus nidulans and Fusarium oxysporum filamentous fungi took the growth inhibition effect of the cell at the concentration of 600ppm, 400ppm, and 100ppm. respectively.

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Decomposition rate of iprobenfos, isoprothiolane, and diazinon by some environmental factors in aqueous systems (몇가지 수중 환경요인에 의한 iprobenfos, isoprothiolane 및 diazinon의 분해속도)

  • Park, Byung-Jun;Choi, Ju-Hyun;Lee, Byung-Moo;Im, Geon-Jae;Kim, Chan-Sub;Park, Kyung-Hun
    • The Korean Journal of Pesticide Science
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    • v.2 no.2
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    • pp.39-44
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    • 1998
  • Three pesticides for paddy rice, iprobenfos, isoprothiolane, and diazinon were examined on some environmental factors, their hydrolysis, microbial degradation, and photolysis in aqueous systems. Iprobenfos was mainly degraded by microorganisms and its half-life was 5.7 days at $28^{\circ}C$ in aqueous systems. Hydrolysis of iprobenfos was accelerated by the higher temperature, but its photodegradation was accelerated by the lower pH. Isoprothiolane was rapidly decomposed by two factors, microorganisms and sunlight. The half-life of isoprothiolane by sunlight was 91 days at pH 9.0, while it was 13 days at pH 4.0 and 16 days at pH 7.2. However, it was shortened under low pH condition. In aqueous system, diazinon was degraded by all of three factors and its degradation rate was remarkably accelerated by acidic solution. Main degradation factors of iprobenfos, isoprothiolane, and diazinon in the aqueous system were investigated by microbial degradation, photolysis, and hydrolysis, respectively. The strains of microbial degradation for iprobenfos, isoprothiolane, and diazinon in the aqueous environment were identified as Pseudomonas putida, Alcaligenes xylosoxydans ss, Klebsiella planticola/ornithinllytica, respectively. The similarity rates of identity were $54.8{\sim}86.2%$ with biolog-system.

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The Characteristics of Imipenem-Resistant Bacteria Isolated from One Patient (한 환자에게서 분리된 Imipenem 내성세균들의 특성)

  • Park, Chul;Lee, Hyeok-Jae;Seo, Min-Young
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.413-419
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    • 2017
  • Four imipenem-resistant bacteria were isolated from the clinical specimens of a patient with pneumonia. To identify the isolates, we used the GN card of Vitek II system and performed a phylogenetic analysis based on 16S rRNA gene sequence. The isolates were identified as P. aeruginosa (2 strains), P. monteilii (1 strain), and P. putida (1 strain), and were tested for antibiotic resistance after determining the MIC of imipenem to be $${\geq_-}8{\mu}g/mL$$ using the AST-N225 card of Vitek II system. The imipenem-resistant genotypes were determined using PCR products amplified using specific ${\beta}-Lactamase$ gene primers. The MBL gene was identified in all four isolates. One strain of P. aeruginosa exhibited the VIM and SHV-1 type genes, while the other strain exhibited both VIM and OXA group II genes. According to the antimicrobial susceptibility test, the bacteria were more susceptible to amikacin than other antibiotics. DNA fingerprint analysis using ERIC-PCR to analyze the epidemiological relationship between strains estimated that both the P. aeruginosa isolates were similar, but exhibited different DNA band types. It is uncommon to find four strains of imipenem-resistant bacteria with different DNA band types in a single patient.

Isolation, Identification and Antagonisms of Rhizospheric Antagonists to Cucumber Wilt Pathogen, Fusarium oxysporum f. sp. cucumerinum Owen (오이 덩굴쪼김병균에 대한 오이 근권길항미생물의 분리, 동정 및 길항작용)

  • Jee Hyeong Jin;Kim Hee Kyu
    • Korean Journal Plant Pathology
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    • v.3 no.3
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    • pp.187-197
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    • 1987
  • Bacteria and fungi antagonistic to Fusarium oxysporum f. sp. cucumerinum Owen were effectively isolated with each of modified Triple Layer Agar (TLA) technique from rhizosphere soil where cucumber had been grown healthily in plastic film house. Three predominant bacterial isolates selected were identified as Pseudomonas fluorescens, and P. putida, Serratia sp. and three fungal isolates were Gliocladium sp. Trichoderma harzianum, and T. viride. Antagonistic bacteria inhibited $26-45\%$ of germination and $41-56\%$ of germ tube elongation of microconidia of F. oxysporum f. sp. cucumerinum on Water Agar (WA). P. fluorescens was the strongest inhibitor. Several my co parasitisms were observed on dual culture of WA between antagonistic fungi and F. oxysporum f. sp. cucumerinum such as coiling, penetration, overgrowing, and lysis. Mycelial lysis of the pathogen was the most severe at pH 4.6, followed by 3.6, 5.6 and 6.6 of the medium in decreasing order. At pH 6.6, mycelia of the pathogen were not conspicuously damaged, however, the antagonistic fungi formed abundant chlamydospores especially Gliocladium sp. T. harzianum revealed the most excellent antagonism in vitro.

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유류분해 미생물을 이용한 세척 유출수 처리에 관한 연구

  • 김형수;최상일;조장환
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2002.04a
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    • pp.287-290
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    • 2002
  • 계면활성제를 이용한 유류 오염토양 복원기술 적용시 발생되는 세척 유출수를 유류분해 미생물로 알려진 Pseudomonas putida Fl, Pseudomonas oleovorans, Acinetobacter calcoaceticus를 이용하여 생물학적으로 처리하는데 필요한 기초 연구를 실시하였다 세종류의 미생물은 본 연구에 사용된 계면활성제(POE$_{5}$, POE$_{14}$)에 대하여 독성이 없는 것으로 판명되었으며, 배양 종료 후 계면활성제 농도에 따른 TPH 분해율은 0.1%, 0.5%, 1%의 계면활성제 농도에서 유사한 분해율을 나타냈으나, 배양 초기에는 0.1%가 0.5%나 1% 농도에 비하여 높은 분해율을 보였다 pH의 변화에 따른 TPH 분해율은 적용 pH 범위(4, 6.5, 9)내에서 모두 생육이 가능하였으며, 세 종류의 미생물은 pH 6.5에서 가장 높은 TPH 분해 율을 나타내었다.

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Homology Analysis Among the Biphenyl and 4-Chlorobiphenyl Degrading Genes by Southern Hybridization (Southern Hybridization에 의한 Biphenyl 및 4-Chlorobiphenyl 분해유전자들의 상동성 분석)

  • 남정현;김치경;이재구;이길재
    • Microbiology and Biotechnology Letters
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    • v.22 no.1
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    • pp.37-44
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    • 1994
  • The homology among the genes coding for degradation of bipheny(BP) and 4-chlorobiphenyl(4CB) was comparatively analyzed by Southern hybridization in several BP/4CB degrading bacterial strains. As the hybridization results of their genomic DNAs with pcbABCD as the DNA probe, the group of Pseudomonas sp. DJ-12. P08 and P27 strain was separated by the group of P20 and P1242 strains. The P. pseudoalcaligenes KF707 showed the hybidization signal which was homologous to the group of DJ-12, but they had different restriction endonuclease sites. The pcbAB genes in pCUl recombinant plasmid from Pseudomonas sp. DJ-12 appeared to be homologous to pchAB genes in pKTF20 cloned from P. pseudoalcaligenes KF707, but the C genes in both strains were not homologous. The bphABC in pKTF20 showed the signals homologous to the cbp ACB in pAW6194 cloned from P. putida OU83, but homologous signal was not found botween the pcbABCD genes in pCUl and the cbpADCB genes in pAW6194 recombbinant plasmid.

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Detection of blaKPC and blaNDM Genes from Gram-Negative Rod Bacteria Isolated from a General Hospital in Gyeongnam (경남지역 종합병원에서 분리된 그람음성막대균으로부터 blaKPC 및 blaNDM 유전자 검출)

  • Yang, Byoung Seon;Park, Ji Ae
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.1
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    • pp.49-59
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    • 2021
  • This study investigated the use of real-time PCR melting curves for the diagnosis of blaKPC and blaNDM genes among the most frequently detected carbapenemase-producing Enterobacteriaceae in Korea. As a means of addressing the shortcomings of phenotype tests and conventional PCR. The modified Hodge test confirmed positivity in 25 of 35 strains, and carbapenemase inhibition testing confirmed positivity in 14 strains by meropenem+PBA or meropenem+EDTA. PCR analysis showed amplification products in 25 strains of Klebsiella pneumoniae carbapenemases (KPC), 10 of K. pneumoniae, 5 of E. coli, 5 of A. baumannii, 4 of P. aeruginosa, and 1 of P. putida. New Delhi metallo β-lactamase (NDM) identified amplification products in 8 strains, that is, 2 K. pneumoniae, 3 E. coli, 1 P. aeruginosa, 1 E. cloacae, and 1 P. retgeri strains. Real-time PCR melting curve analysis confirmed amplification in 25 strains of KPC and 8 strains of NDM, and these results were 100% consistent with PCR results. In conclusion, our findings suggest early diagnosis of carbapenem resistant Enterobacteriaceae by real-time PCR offers a potential means of antibacterial management that can prevent and control nosocomial infection spread.

Biotransformation of Flavone by CYP105P2 from Streptomyces peucetius

  • Niraula, Narayan Prasad;Bhattarai, Saurabh;Lee, Na-Rae;Sohng, Jae Kyung;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.22 no.8
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    • pp.1059-1065
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    • 2012
  • Biocatalytic transfer of oxygen in isolated cytochrome P450 or whole microbial cells is an elegant and efficient way to achieve selective hydroxylation. Cytochrome P450 CYP105P2 was isolated from Streptomyces peucetius that showed a high degree of amino acid identity with hydroxylases. Previously performed homology modeling, and subsequent docking of the model with flavone, displayed a reasonable docked structure. Therefore, in this study, in a pursuit to hydroxylate the flavone ring, CYP105P2 was co-expressed in a two-vector system with putidaredoxin reductase (camA) and putidaredoxin (camB) from Pseudomonas putida for efficient electron transport. HPLC analysis of the isolated product, together with LC-MS analysis, showed a monohydroxylated flavone, which was further established by subsequent ESI/MS-MS. A successful 10.35% yield was achieved with the whole-cell bioconversion reaction in Escherichia coli. We verified that CYP105P2 is a potential bacterial hydroxylase.

Binding Site of Heavy Metals in the Cell of Heavy Metal-Tolerant Microorganisms (중금속 내성균의 세포내 중금속 결합 위치)

  • Cho, Ju-Sik;Lee, Hong-Jae;Lee, Young-Han;Sohn, Bo-Kyoon;Jung, Yeun-Kyu;Heo, Jong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.17 no.3
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    • pp.246-253
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    • 1998
  • Heavy metal-tolerant microorganisms, such as Pseudomonas putida, P. aeruginosa, P. chlororaphis and P. stutzeri which possessed the ability to accumulate cadmium, lead, zinc and copper, respectively, were isolated from industrial wastewaters and mine wastewaters polluted with various heavy metals. The binding sites of heavy metal in the cells were investigated by chemical modification of functional groups the cell walls. To determine the binding sites of heavy metal in the cells, electrochemical charge of amine and carboxyl groups in the cell walls of heavy metal-tolerant microorganisms were chemically modified. Chemical modifications of amine groups did not affect the heavy metal uptake as compared to native cell walls. In contrast, modifications of carboxyl groups drastically decreased heavy metal uptake as compared to native cell walls, and electron microscopy confirmed that the form and structure of the heavy metal uptake were different from those of native cell walls. The results suggested that the carboxyl groups were the major sites of heavy metal uptake in the heavy metal-tolerant microorganism cell.

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