• Title/Summary/Keyword: P. putida

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Cloning and Expression of pcbC and pcbD Genes Responsible for 2,3-Dihydroxybiphenyl Degradation from Pseudomonas sp. P20

  • Nam, Jung-Hyun;Oh, Hee-Mock;Kim, Chi-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.5 no.2
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    • pp.68-73
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    • 1995
  • Pseudomonas sp. P20 was shown to be capable of degrading biphenyl and 4-chlorobiphenyl (4CB) to produce the corresponding benzoic acids wnich were not further degraded. But the potential of the strain for biodegradation of 4CB was shown to be excellent. The pcbA, B, C and D genes responsible for the aromatic ring-cleavage of biphenyl and 4CB degradation were cloned from the chromosomal DNA of the strain. In this study, the pebC and D genes specifying degradation of 2, 3-dihydroxybiphenyl (2, 3-DHBP) produced from biphenyl by the pebAB-encoded enzymes were cloned by using pBluescript SK(+) as a vector. From the pCK102 (9.3 kb) containing pebC and D genes, pCK1022 inserted with a EcoRI-HindIII DNA fragment (4.1 kb) carrying pebC and D and a pCK1092 inserted with EcoRI-XbaI fragment (1.95 kb) carrying pebC were constructed. The expression of pcbC and D' in E. coli CK102 and pebC in E. coli CK1092 was examined by gas chromatography and UV-vis spectrophotometry. 2.3-dihydroxybiphenyl was readily degraded to produce meta-cleavage product (MCP) by E. coli CK102 after incubation for 10 min, and then only benzoic acid(BA) was detected in the 24-h old culture. The MCP was detected in E. coli CK1022 containing pebC and 0 genes (by the resting cells assay) for up to 3 h after incubation and then diminished completely in 8 h, whereas the MCP accumulated in the E. coli CK1092 culture even after 6 h of incubation. The 2, 3-DHBP dioxygenases (product of pebC gene) produced by E. coli CK1, CK102, CK1023, and CK1092 strains were measured by native PAGE analysis to be about 250 kDa in molecular weight, which were about same as those of Pseudomonas sp. DJ-12, P. pseudoa1caligenes KF707, and P. putida OU83.

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Enhancement of Plant Growth and Suppression of Damping-off of Cucumber by Low Temperature Growing Pseudomonas fluorescens Isolates (저온 생장성 Pseudomonas fluorescens M45와 MC07을 이용한 오이의 생육촉진과 모잘록병의 방제)

  • 염주립;박창석
    • Korean Journal Plant Pathology
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    • v.11 no.3
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    • pp.252-257
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    • 1995
  • Growth rates of the low temperature growing isolates, Pseudomonas fluorescens M45 and MC07, reached maximum stationary phase within 50 hrs at the low temperature, 4$^{\circ}C$. But an ordinary biocontrol agent P. putida Pf3 did not reach logarithmic growth phase until 80 hrs. The culture filtrates of M45 and MC07 significantly inhibited the mycelial growths of Pythium ultimum, Rhizoctonia solani and Phytophthora capsici. Detached cotyledons of cucumber grown on Murashige and Skoog agar medium were much enhanced in their growth, compared to those without the filtrates. Population densities of M45 and MC07 in the rhizosphere at 14$^{\circ}C$ were more stable than at 27$^{\circ}C$. When M45 and MC07 were treated into soil, the population density of MC07 continuously increased up to 9 days after treatment, and sustained the initial inoculum density up to 60 days. Cucumber damping-offs caused by P. ultimum and R. solani were significantly reduced by applying M45 as seed-inoculant and by soil treatment with MC07. The combined treatment of M45 and MC07 provided greater effect in reducing the disease incidence than that obtained by single treatments.

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Cytotoxic and Antimicrobial Activities of Bioactive Monoterpenophenols

  • Oh In Kio;Lee Hyun Ok;Ahn Jong Woong;Kim Hyung Min;Shin Ji Hee;Lim Jin A;Chun Hyun Ja;Baek Seung Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.6
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    • pp.1270-1276
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    • 2002
  • Compounds 1 - 12 were tested for their growth inhibitory effects against tumor cell lines using two different 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and sulforhodamine B protein (SRB) assays and antimicrobial activity. The cytotoxic activity of methyl-4-[{(2E)-3,7-dimethyl-2,6-octadienyl}oxy]-3-methoxy benzoate (1) exhibit more active than that of 5-fluorouracil (11) on human oral epithelioid carcinoma (KB, ATCC No. OCL 17) cell lines. But this compound (1) on human skin melanoma (SK-MEL-3, HBT 69) cell lines shows less active than that of adriamycin (12). However, compound 9 showed the antimicrobial activity against S. epidermidis (MIC, 15.625 ㎍/㎖), S. aureus, C. albicans (MIC, 31.25 ㎍/㎖), S. mutans, S. typhimurium, P. putida (MIC. 125 ㎍/㎖) and P. aeruginosa (MIC, 500 ㎍/㎖).

Screening of Cytotoxicity and Antimicrobial Effects of Hexane Extracts from Cornis fructus (산수유 헥산 추출물의 항균효과 및 세포독성)

  • Chun Hyun Ja;Choi Won Hyung;Lee Jeong Ho;Yang Hyun Ok;Baek Seung Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.476-480
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    • 2003
  • Cornis fructus was extracted by successive extraction and then fractionated with hexane extract to get active fractions. This study was performed to determine the cytotoxic effect of hexane extract from Cornis fructus on NIH 3T3 fibroblasts and cancer cell lines using MTT assay. Hexane extract showed cytotoxic effect against A549, B16 melanoma and MDA-MB-231. Futher fractionation with hexane extract was performed to obtain effective fraction, fraction 3 showed the cytotoxic effect against A549 and MDA-MB-231. In antimicrobial test of each fraction of hexane extract, fraction 5 showed antimicrobial activities against P. putida and P. aeruginosa.

Effects of Plants, Rhizobacteria and Physicochemical Factors on the Phytoremediation of Contaminated Soil (오염 토양의 식물상 복원효율에 미치는 식물, 근권세균 및 물리.화학적 인자의 영향)

  • Hong, Sun-Hwa;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.261-271
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    • 2007
  • Phytoremediation is an economic and environmentally friendly technique to remediate contaminated-soil. In this study, the effects of plants, rhizobacteria and physicochemical factors on phytoremediation have been reviewed. For successful phytoremediation, the selection of plants is primarily important. To remediate soil contaminated with petroleum hydrocarbon, raygrass (Lolium multiflorum lam), white mustard, vetch (Vicia villosa), tall fescue (Festuca arundinacea), legumes, poplar, and Pine (Pinus densiflora) were mainly applied, and the removal efficiency of petroleum hydrocarbon were ranged 68 to 99%. Corn (Zea mays), raygrass (Lolium multiflorum lam), vetch (Vicia villosa), mustard, clover (Trifolium repens), and tall fescue (Festuca arundinacea) were used for the removal of polycyclic aromatic hydrocarbon, and their removal efficiencies were 50-98%. Rhizobacteria play significant roles for phytoremediation because they can directly participate in the degradation of contaminant as well as promoting plants growth. The following rhizobacteria were preferred for phytoremediation: Azospirillum lipoferum, Enterobactor cloacae, Azospirillum brasilense, Pseudomonas putida, Burkholderia xenovorans, Comamonas testosterone, Pseudomonas gladioli, Azotobacter chroococcum, Bacillus megaterium, and Bacillus subtilis. Pysicochemical factors such as pH, temperature, nutrient, electron acceptor, water content, organic content, type of contaminants are consequential limiting factors for phytoremediation.

Effect of Bioluminescence Stimulating Agent of the Genetically Engineered Strain KG1206 on the Monitoring of the Petroleum Hydrocarbon Contaminated Groundwater Samples (발광유전자 재조합 균주 활성 촉진 조건이 석유계 탄화수소 오염지하수 모니터링에 미치는 영향)

  • Ko, Kyung-Seok;Kong, In-Chul
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.1
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    • pp.79-84
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    • 2008
  • This paper describes the application of bioluminescence stimulating agents on a genetically engineered microorganism, Pseudomonas putida mt-2 KG1206, to monitor toluene analogs using in groundwater samples from petroleum hydrocarbon contaminated sites. The maximum bioluminescent response with pure chemicals followed in the order: m-methyl benzyl alchohol > m-toluate > toluene > m-xylene > benzoate > p-xylene > o-xylene. Generally, the bioluminescence production of strain mixed with groundwater samples was dependent on the contaminated total inducer concentrations. However, few samples showed opposite results, where these phenomena may be caused by the complexicity of environmental samples. Two chemicals, SL(sodium lactate) and KNO$_3$, were tested to determine a better bioluminescence stimulant. Both chemicals stimulate the bioluminescence activity of strain KG1206, however, a slightly high bioluminescence was observed with nitrogen chemical. This selected stimulant was then tested on samples collected from contaminated groundwater samples. The bioluminescence activity of all samples mixed with the strain was stimulated with KNO$_3$ amendment. This suggests that the low bioluminescence activity exhibited by the environmental groundwater samples can be stimulated by amending the culture with a proper agent, such as nitrogen compound. These findings would be useful, especially, when strain was used to monitor the groundwater samples contaminated with low inducer contaminants. Overall, the results of this study found the ability of bioluminescence producing bacteria to biosensor a specific group of environmental contaminants, and suggest the potential for more efficient preliminary application of this engineered strain in a field-ready bioassay.

Biocontrol of Phytophthora Blight and Anthracnose in Pepper by Sequentially Selected Antagonistic Rhizobacteria against Phytophthora capsici

  • Sang, Mee Kyung;Shrestha, Anupama;Kim, Du-Yeon;Park, Kyungseok;Pak, Chun Ho;Kim, Ki Deok
    • The Plant Pathology Journal
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    • v.29 no.2
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    • pp.154-167
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    • 2013
  • We previously developed a sequential screening procedure to select antagonistic bacterial strains against Phytophthora capsici in pepper plants. In this study, we used a modified screening procedure to select effective biocontrol strains against P. capsici; we evaluated the effect of selected strains on Phytophthora blight and anthracnose occurrence and fruit yield in pepper plants under field and plastic house conditions from 2007 to 2009. We selected four potential biocontrol strains (Pseudomonas otitidis YJR27, P. putida YJR92, Tsukamurella tyrosinosolvens YJR102, and Novosphingobium capsulatum YJR107) among 239 bacterial strains. In the 3-year field tests, all the selected strains significantly (P < 0.05) reduced Phytophthora blight without influencing rhizosphere microbial populations; they showed similar or better levels of disease suppressions than in metalaxyl treatment in the 2007 and 2009 tests, but not in the 2008 test. In the 2-year plastic house tests, all the selected strains significantly (P < 0.05) reduced anthracnose incidence in at least one of the test years, but their biocontrol activities were variable. In addition, strains YJR27, YJR92, and YJR102, in certain harvests, increased pepper fruit numbers in field tests and red fruit weights in plastic house tests. Taken together, these results indicate that the screening procedure is rapid and reliable for the selection of potential biocontrol strains against P. capsici in pepper plants. In addition, these selected strains exhibited biocontrol activities against anthracnose, and some of the strains showed plant growth-promotion activities on pepper fruit.

Chloroplast-type Ferredoxin Involved in Reactivation of Catechol 2,3-Dioxygenase from Pseudomonas sp.S-47

  • Park, Dong-Woo;Chae, Jong-Chan;Kim, Young-Soo;Iida, Toshiya;Kudo, Toshiaki;Kim, Chi-Kyung
    • BMB Reports
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    • v.35 no.4
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    • pp.432-436
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    • 2002
  • Pseudomonas sp. S-47 is capable of degrading catechol and 4-chlorocatechol via the meta-cleavage pathway. XyITE products catalyze the dioxygenation of the aromatics. The sylT of the strain S-47 is located just upstream of the xylE gene. XylT of the strain S-47 is located just upstream of the xylE gene. XyIT is typical chloroplast-type ferredoxin, which is characterized by 4 cystein residues that are located at positions 41, 46, 49, and 81. The chloroplast-type ferredoxin of Pseudomonas sp. S-47 exhibited a 98% identity with that of P. putida mt-2(TOL plasmid) in the amino acid sequence, but only about a 40 to 60% identity with the corresponding enzymes from other organisms. We constructed two recombinant plasmids (pRES1 containing xylTE and pRES101 containing xylE without xylT) in order to examine the function of XyIT for the reactivation of the catechol 2,3-dioxygenase (XyIE) that is oxidized with hydrogen peroxide was recovered in the catechol 2,3-dioxygenase (C23O) activity about 4 mimutes after incubation, but the pRES101 showed no recovery. That means that the typical chloroplast-type ferredoxin (XyIT) of Pseudomonas sp. S-47 is involved in the reactivation of the oxidized C23O in the dioxygenolytic cleavage of aromatic compounds.

Cytotoxicity and antimicrobial effects of the methanolic extract of Sophora flavescens Ait. (IV)

  • Baek, Seung-Hwa;Kang, Kil-Ung;Lee, Jeong-Ho;Park, Nang-Kyu;Chai, Kyu-Yun;You, Il-Soo;Kim, Jong-Soo;Ryu, Do-Gon;Lee, Kang-Min;Yang, Eun-Yeong;Lee, Hyun-Ok
    • Advances in Traditional Medicine
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    • v.1 no.2
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    • pp.45-51
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    • 2000
  • This study was carried out to evaluate cytotoxicity of the methanol extract from Sophora flavescens Ait. against L1210 (lymphocytic leukemia) and $P388D_1$ (lymphoid neoplasma) Cells in vitro. We have determined cytotoxicity by the MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H- tetrazolium bromide) assay. The order of cytotoxicity of Sophora flavescens Ait. extracts against L1210 and $P388D_1$ cells in vitro is as follows: Fr. 4 > Fr. 3 > Fr. 5 > Fr. 2 > Fr. 1. These results suggest that the fraction 4 of the methanol extracts from Sophora flavescens Ait. may be a valuable choice for the development of antitumor agents. In order to develop an antimicrobial agent, dried Sophora flavescens Ait. was extracted with hot methanol, and then antimicrobial activity (MIC test) was investigated. In this study, the fraction 3 of the methanol extracts from the roots of S. flavescens showed strong growth inhibition activity against gram-positive and gram-negative bacteria (MIC, $3.125\;{\mu}g/ml$) such as S. mutans, S. epidermidis and P. putida. These results indicate that fractions 3 and 4 inhibit tumor cells and bacteria.

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Meta-analysis Reveals That the Genus Pseudomonas Can Be a Better Choice of Biological Control Agent against Bacterial Wilt Disease Caused by Ralstonia solanacearum

  • Chandrasekaran, Murugesan;Subramanian, Dharaneedharan;Yoon, Ee;Kwon, Taehoon;Chun, Se-Chul
    • The Plant Pathology Journal
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    • v.32 no.3
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    • pp.216-227
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    • 2016
  • Biological control agents (BCAs) from different microbial taxa are increasingly used to control bacterial wilt caused by Ralstonia solanacearum. However, a quantitative research synthesis has not been conducted on the role of BCAs in disease suppression. Therefore, the present study aimed to meta-analyze the impacts of BCAs on both Ralstonia wilt disease suppression and plant (host) growth promotion. The analysis showed that the extent of disease suppression by BCAs varied widely among studies, with effect size (log response ratio) ranging from -2.84 to 2.13. The disease incidence and severity were significantly decreased on average by 53.7% and 49.3%, respectively. BCAs inoculation also significantly increased fresh and dry weight by 34.4% and 36.1%, respectively on average. Also, BCAs inoculation significantly increased plant yield by 66%. Mean effect sizes for genus Pseudomonas sp. as BCAs were higher than for genus Bacillus spp. Among antagonists tested, P. fluorescens, P. putida, B. cereus, B. subtilis and B. amyloliquefaciens were found to be more effective in general for disease reduction. Across studies, highest disease control was found for P. fluorescens, annual plants, co-inoculation with more than one BCA, soil drench and greenhouse condition were found to be essential in understanding plant responses to R. solanacearum. Our results suggest that more efforts should be devoted to harnessing the potential beneficial effects of these antagonists, not just for plant growth promoting traits but also in mode of applications, BCAs formulations and their field studies should be considered in the future for R. solanacearum wilt disease suppression.