• Title/Summary/Keyword: Burkholderia sp.

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Comparative Analysis of Aniline Dioxygenase Genes from Aniline Degrading Bacteria, Burkholderia sp. HY1 and Delftia sp. HY99. (Aniline 분해균주 Burkholderia sp. HY1과 Delftia sp. HY99에서 유래된 Aniline Dioxygenases 유전자의 비교 분석)

  • Kahng, Hyung-Yeel;Oh, Kye-Heon
    • Microbiology and Biotechnology Letters
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    • v.35 no.2
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    • pp.104-111
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    • 2007
  • In this study, aniline dioxygenase genes responsible for initial catabolism of aniline in Burkholderia sp. HY1 and Delftia sp. HY99 were cloned and the amino acid sequences were comparatively analyzed, which already have been reported as bacteria utilizing aniline as a sole source of carbon and nitrogen, B. sp. HY1 was found to have at least a plasmid, and the plasmld-cured strain, B. sp. HY1-PC obtained using mitomycin C was tested with wild type strain to investigate whether the former maintained the degradability for aniline. This proved that the aniline oxygenase gene from B. sp. HY1 was located in chromosomal DNA, not in plasmid DNA. Aniline dioxygenase small subunits from B. sp. HY1 and D. sp. HY99 were found, based on 146 amino acids, to share 79% similarity. Notably, ado2 genes from B. sp. HY1 and D. sp. HY99 which were found to be terminal dioxygenase of aniline dioxygenase small subunit showed 99% similarity in the deduced amino acid sequences with tdnA2 of Frateuria sp. ANA-18 and danA2 of D. sp. AN3, respectively. Besides, enzyme assay and amino acid sequence analysis of catechol dioxygenase supported the previous report that B. sp. HY1 might occupy ortho-cleavage pathway using catechol 1,2-dioxygenase, while D. sp. HY99 might occupy catechol 2,3-dioxygenase for meta-cleavage pathway.

Molecular Characterization of Burkholderia Strains Isolated from Rice Cultivars (Oryza sativa L.) for Species Identification and Phylogenetic Grouping

  • Madhaiyan, Munusamy;Poonguzhali, Selvaraj;Kwon, Soon-Wo;Song, Myung-Hee;Sa, Tong-Min
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1005-1010
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    • 2008
  • The genus Burkholderia consists of extremely versatile bacteria that occupy diverse niches and are commonly encountered in the rhizosphere of crop plants. In this study, we characterized three plant growth promoting strains assigned as Burkholderia sp. using biochemical and molecular characterization. The Burkholderia spp. strains CBMB40, CBPB-HIM, and CBPB-HOD were characterized using biochemical tests, BIOLOG carbon substrate utilization, fatty acid methyl ester analysis, analysis of recA gene sequences, and DNA-DNA hybridization. The results from these studies indicated that the strains CBMB40, CBPB-HIM, and CBPB-HOD can be assigned under Burkholderia vietnamiensis, Burkholderia ubonensis, and Burkholderia pyrrocinia, respectively.

Degradation of 3-Methyl-4-nitrophenol, a Main Product of the Insecticide Fenitrothion, by Burkholderia sp. SH-1 Isolated from Earthworm (Eisenia fetida) Intestine

  • Kim, Seon-Hwa;Park, Myung-Ryeol;Han, Song-Ih;Whang, Kyung-Sook;Shim, Jae-Han;Kim, In-Seon
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.281-287
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    • 2007
  • Microorganisms were isolated from earthworm intestine and examined for their ability to degrade 3-methyl-4-nitrophenol (MNP), a main degradation product of the insecticide fenitrothion. An isolate that showed the best degradation of MNP was selected for further study. The 16S rRNA analysis showed that the isolate belongs to the genus of Burkholderia, close to phenanthrene-degrading Burkholderia sp. S4.9, and is named Burkholderia sp. SH-1. When time-course degradation of MNP by SH-1 was examined by high performance liquid chromatographic analysis, almost complete degradation of MNP was observed within 26 h. Colony forming unit value assays indicated that the isolate SH-1 was capable of utilizing MNP as a sole carbon source. SH-1 could also degrade p-nitrophenol (PNP) but could not degrade ortho-substituted nitroaromatics such as 2,4-, 2,6- and 2,5-dinitrophenol. Catechol was detected as the main degration product of MNP and PNP. SH-1 was also found in the soil from which earthworms were obtained. These results suggest that the dispersal of Burkholderia sp. SH-1 into different environment with the aid of earthworms is likely to play a role in bioremediation of the soil contaminated with MNP.

Acute Oral, Pulmonary and Intravenous Toxicity/Pathogenicity Testing of Burkholderia pyrrocinia CAB08106-4 of in Rats (랫드를 이용한 Burkholderia pyrrocinia CAB08106-4의 급성경구, 호흡기, 정맥독성/병원성시험)

  • Kwon, Min;Kang, Tae-Ku;Chung, Chang-Kook;Park, Cheol-Beom
    • The Korean Journal of Pesticide Science
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    • v.17 no.3
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    • pp.193-199
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    • 2013
  • Burkholderia pyrrocinia CAB08106-4 has an anti-bacterial effect on Garlic White Rot caused by Sclereotium cepivorum and Sclereotium sp.. It is an environmentally friendly microbial product that prevents and controls a variety of phytopathogens involving Garlic White Rot caused by Sclereotium cepivorum and Sclereotium sp.. The aim of this study was to assess and to compare the pathogenicity of Burkholderia pyrrocinia CAB08106-4 by single exposure of rats through several routes such as oral, intranasal and intravenous. For the acute toxicity / pathogenicity study, the animals were sacrificed on days 3, 7, 14 and 21, and macroscopically observed their organs to examine the numbers of internally-retained pesticidal microbes. Clinical examinations were performed daily during administration period, and body weight gain was evaluated. In the study, no clinical sign, weight gain and mortality were observed in relation to the administration of test article. The significant changes of internal/external microbes by test article were not detected. The pathological findings in relation to the administration of the test article in the necropsy were not observed. It could be concluded that the microorganism was not toxic or pathogenic in rats via oral, intranasal and intravenous route.

Antimicrobial activities of Burkholderia sp. strains and optimization of culture conditions (Burkholderia sp. OS17의 항균활성 증진을 위한 배양최적화)

  • Nam, Young Ho;Choi, Ahyoung;Hwang, Buyng Su;Chung, Eu Jin
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.428-435
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    • 2018
  • In this study, we isolated and identified bacteria from freshwater and soil collected from Osang reservoir, to screen antimicrobial bacteria against various pathogenic bacteria. 38 strains were isolated and assigned to the class Proteobacteria (22 strains), Actinobacteria (7 strains), Bacteroidets (6 strains), and Firmicutes (3 strains) based on 16S rRNA gene sequence analysis. Among them, strain OS17 showed a good growth inhibition against 5 methicillin-resistant Staphylococcus aureus subsp. aureus strains and Bacillus cereus, Bacillus subtilis, Filobasidium neoformans. As a result of the 16S rRNA gene sequence analysis, strain OS17 show the high similarity with Burkholderia ambifaria $AMMD^T$, B. diffusa $AM747629^T$, B. tettitorii $LK023503^T$ 99.8%, 99.7%, 99.6%, respectively. We investigated cell growth and antimicrobial activity according to commercial culture medium, temperature, pH for culture optimization of strain OS17. Optimal conditions for growth and antimicrobial activity in strain OS17 were found to be: YPD medium, $35^{\circ}C$ and pH 6.5. When the strain was cultured in LB, NB, TSB, R2A media at $20^{\circ}C$ and $25^{\circ}C$, the antimicrobial activity did not show. Culture filtrate of strain OS17 showed antimicrobial activity against 5 MRSA strains, Bacillus cereus, Bacillus subtilis, and Filobasidium neoformans with inhibition zones from 2 to 8 mm. Optimal reaction time was 48 h in YPD medium, 100 rpm and 0.3 vvm in 2 L-scale fed-batch fermentation process for antimicrobial activity. Culture optimization of strain OS17 can be improved on antimicrobial activity. Therefore, the antimicrobial activity of Burkholderia sp. OS17 had potential as antibiotics for pathogens including MRSA.

Isolation and Identification of a Marine Bacterium Producing an Immunostimulant (면역증강물질을 생산하는 해양미생물의 분리 및 동정)

  • 최혜정;정명주;정영기
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.6
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    • pp.1145-1150
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    • 2000
  • 면역증강물질을 생산하는 균주를 해수로부터 분리하여 동정한 결과 크림색의 둥근 점질성의 집략을 나타내었으며, Gram 음성의 간산형이고, 미약한 운동성을 가지며 catalase 양성과 oxidase 음성 반응을 보였다. 균주를 6시간 , 20시간, 72시간, 및 144시간 뱅양하여 전자 현미경사에서 관찰한 결과 20시간 배양한 영양세포는 1.25~0.6$\times$0.6$\mu\textrm{m}$ 크기의 간균 형태를 갖추었으며 시간이 흐를수록 세포내 관립의 밀도가 증가하면서 세포형태가 다형태성으로 변하였다. 세포내 과립의 존재를 확인하기 위해 sudan black B로 염색한 결과 양성으로 polyhy-droxy butyrate로 예상되었다. 생육을 위한 최적 온도는 3$0^{\circ}C$, pH는 3.0~10.0이었으며, 호기성균이었다. D-Glu-cose, D-mannose, D-mannitiol. insitol, maltose 등의 당과 L-asparagine, L-glutamate 등의 아미노산을 이용하였고 특시, O/F test에서 glucose와 maltose를 산화 하였다. 이상과 같은 결과로 해양세균 유래인 Pseudo-mallei group의 Burkholderia 속으로 확인되었음로 본 균주를 Burkholderia sp. IS-203으로 명명하였다.

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Degradation of Phenanthrene and Pyrene by Burkholderia sp. D5 (Burkholderia sp. D5에 의한 phenanthrene과 pyrene 분해)

  • Kim, Tae-Jeong;Jo, Gyeong-Suk;Ryu, Hui-Uk
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.267-271
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    • 2003
  • Burkholderia sp. D5, a polyaromatic hydrocarbons(PAHs)-degrading bacterium, was isolated from oil-contaminated soil. The bacterium could utilize phenanthrene (Phe) as a sole carbon source but could not use pyrene (Pyr). However, the strain could degrade Pyr when a cosubstrate such as yeast extract (YE) was supplemented. The PAH degradation rate of the bacterium was enhanced by the addition of other organic materials such as YE, peptone and glucose. YE was a particularly effective additive in stimulating cell growth as well as PAH degradation. When 1 g-YE/L was supplemented into the basal salt medium (BSM) with 215 mg-Phe/L, the specific growth rate (0.28 h-1) and Phe-degrading rate (29.30 μmol/L/h) were enhanced approximately ten and two times more than those obtained in the BSM with 215 mg-Phe/L, respectively. Through kinetic analysis, the maximum specific growth rate (μmax) and PAH degrading rate (Vmax) for Phe were obtained as 0.34/h and 289 ${\mu}mol$/L/h, respectively. Also, μmax and Vmax for Pyr were 0.27 h-1 and 50 ${\mu}mol$/L/h, respectively. The degradation rates for each Phe (2.20 μmol/L/h) and Pyr (2.18 μmol/L/h) were lower in mixture substrates than in a single substrate (29.30 ${\mu}mol$/L/h and 9.58 ${\mu}mol$/L/h, respectively). Burkholderia sp. D5 can degrade Phe and Pyr contained in soil, and the PAH degradation rates in soil were 20.03 ${\mu}mol$/L/h for Phe and 1.09 ${\mu}mol$/L/h for Pyr.

Characterization of PAH-Degrading Bacteria from Soils of Reed Rhizosphere in Sunchon Bay Using PAH Consortia (순천만 갈대근권 토양으로부터 얻은 PAH 분해세균의 특성 분석)

  • Kim Sung-Hyun;Kang Sung-Mi;Oh Kye-Heon;Kim Seung-Il;Yoon Byoung-Jun;Kahng Hyung-Yeel
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.208-215
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    • 2005
  • This study was accomplished in order to collect fundamental data on microbial roles in recycling process of reed rhizosphere. Sunchon bay, which is considered as one of the marsh and mud environments severely affected by human activities such agriculture and fisheries, was selected as a model place. In our initial efforts, two bacterial consortia were obtained by enrichment culture using PAH mixtures containing anthracene, naphthalene, phenanthrene and pyrene as the sources of carbon and energy, and four pure bacteria capable of rapid degradation of PAH were isolated from them. Four strains designated as SCB1, SCB2, SCB6, and SCB7 revealed by morphological, physiological and molecular analyses were identified as Burkholderia anthina, Alcaligenes sp., Achromobacter xylosoxidans., and Pseudomonas putida, respectively with over $99{\%}$ confidence. Notably, Burkholderia anthina SCB1 and Alcaligenes sp. SCB2 were found to utilize anthracene and pyrene more quickly than naphthalene and phenanthrene, whereas Achromobacter xylosoxidans SCB6 and Pseudomonas putida SCB7 exhibited similar growth and degradation patterns except for pyrene. These facts suggest that the rhizosphere microorganisms capable of PAH degradation might be used to clean up the contamination sites with polycyclic aromatic hydrocarbons.

An Enantioselective Amidase from Burkholderia multivorans for the Stereoselective Synthesis of Esfenvalerate

  • Lee, Sang-Hyun;Park, Oh-Jin;Shin, Hyun-Jae
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.936-942
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    • 2014
  • Using racemic (R,S)-2-(4-chlorophenyl)-3-methylbutyramide, an intermediate for the chiral pyrethroid insecticide Esfenvalerate, as a sole nitrogen source in a minimal medium, several strains with high enatioselectivity (${\geq}98%$) were isolated by enrichment techniques. One of the strains, LG 31-3, was identified as Burkholderia multivorans, based on physiological and morphological tests by a standardized Biolog station for carbon source utilization. A novel amidase was purified from B. mutivorans LG 31-3 and characterized. The enzyme exhibited (S)-selective amidase activity on racemic (R,S)-2-(4-chlorophenyl)-3-methylbutyramide. Addition of the racemic amide induced the production of the enantioselective amidase. The molecular mass of the amidase on SDS-PAGE analysis was shown to be 50 kDa. The purified amidase was subjected to proteolytic digestion with a modified trypsin. The N-terminal and internal amino acid sequences of the purified amidase showed a high sequence homology with those deduced from a gene named YP_366732.1 encoding indole acetimide hydrolase from Burkholderia sp. 383.

Optimization of the Production of an Immunostimulant from a Marine Bacterium (해양미생물로부터 면역증강물질의 생산 최적화)

  • 최혜정;정명주;정영기
    • Journal of Life Science
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    • v.12 no.6
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    • pp.759-764
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    • 2002
  • A halophilic bacterium for the production of the immunostimulant was isolated from domestic marine, it was identified as Burkholderia sp. IS-203. The optimal conditions for the production of the immunostimulant were 1 % dextrose and 1 % yeast extract in artificial sea water for carbon and nitrogen sources, respectively. The initial pH and growth temperature for the prodution were 8.0 and $30^{\circ}C$ under the presence of oxygen, respectively.