• Title/Summary/Keyword: Burkholderia cepacia

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Immobilization of Burkholderia cepacia Lipase on Weak Base Styrene Resin Using Polyethyleneimine with Cross-linking (PEI(Polyethyleneimine)를 이용하여 음이온계 레진에 고정화된 Lipase AH 제조 및 효소적 Interesterification을 통한 반응 특성 연구)

  • Lee, Chi Woo;Lee, Ki Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.7
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    • pp.1025-1035
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    • 2014
  • This study assessed the effect of immobilized lipase on weak base styrene resin using polyethyleneimine (PEI) with cross-linking. Two procedures were used in this study. The first one, "mono-layer" lipase immobilization, involves washing PEI after adsorption. The second procedure, "multi-layer" lipase immobilization, has no washing before the cross-linking step. Treverlite XS-100200 (weak base styrene resin) was immersed with PEI solution (2.2 mg/mL). Lipase AH (from Burkholderia cepacia) was adsorbed onto the support coated with PEI before cross-linking with glutaraldehyde. Structured lipid was synthesized by immobilized lipase-catalyzed interesterification using canola oil, palmitic ethyl ester (PEE), and stearic ethyl ester (StEE). Total fatty acid contents of triacylglycerol (TAG) in structured lipids were analyzed to investigate activity, properties, and reusability of immobilized lipases. Activities of immobilized lipases on the multi-layer and mono-layer increased at a high concentration (8 mg/mL) of lipase solution used for immobilization. The results show that immobilized lipase with the mono-layer method at pH 8.0 on resin had the highest total saturated fatty acid content (26.17 area%). Activity of immobilized lipase with the multi-layer method at pH 7.5 on support was lower than that of the mono-layer, but total saturated fatty acid content was 16.79 area% higher than that of lipase AH (15.01 area%).

Expression of human lactoferrin N-lobe in Pichia pastoris and its antibacterial activity (Pichia pastoris에서 사람 락토페린 N-lobe의 발현과 항균활성)

  • Won, Su-Jin;Jo, Jae-Hyung;Kim, Seung-Hwan;Kwon, Hyuk-Jin;Lee, Hyune-Hwan
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.271-279
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    • 2015
  • Lactoferrin (LF) is a multifunctional, iron-binding glycoprotein found in physiological secretions of mammals. LF shows antibacterial, antiviral and antifungal activities. In the present study, a gene encoding the N-terminal lobe of human lactoferrin (hLF) was isolated, cloned and expressed in methylotrophic yeast, Pichia pastoris. The recombinant hLF-N (rhLF-N) protein was secreted into the culture medium at the level of $458{\mu}g/ml$ in 3 L fermentor. The size of purified hLF-N was estimated as 35 kDa when analyzed by SDS-PAGE and western blotting. The rhLF-N was further confirmed by immunodiffusion using the anti-hLF polyclonal antibody. The expression profile analysis by qRT-PCR showed that the relative mRNA expression of rhLF-N was maximal after 2-3 days of methanol induction and reduced gradually at 4 days. The purified rhLF-N showed broad antibacterial activities against the pathogens such as Staphylococcus aureus, E. coli, Pseudomonas aeruginosa, Burkholderia cepacia, and Salmonella typhimurium. However, rhLF-N showed relatively lower activity when compared to peptides derived from LF. In spite of this weak activity, the rhLF-N expressed in P. pastoris might be more advantageous for the industrial application, because rhLF-N is secreted into the culture medium and the production can also be increased by optimization of culture conditions.

Distribution and Characteristics of Acidotolerant Heterotrophic and Naphthalene­Degrading Bacteria in Acidic Soils (산성토양에서 내산성 종속영양세균과 나프탈렌분해세균의 분포 및 특성)

  • Moon Yong-Suk;Chu Kwang-Il;Kim Jongseol
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.313-319
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    • 2004
  • The distribution and characteristics of acidotolerant heterotrophic and naphthalene-degrading bacteria were investigated in two forest areas, one near Ulsan petrochemical industrial complex (Sunam) and the other in countryside (Daeam). Average values of soil pH at Sunam and Daeam were 3.8 and 4.6, respectively. When het­erotrophic and naphthalene-degrading bacteria were enumerated by most probable number (MPN) procedures at Sunam, the median values of heterotrophs growing at pH 7.0 and pH 4.0 were $5.3{\times}10^7\;and\;3.3{times}10^7$ MPN/g, whereas those of naphthalene-degraders were $5.6{\times}10^4\;and\;4.0{times}10^5$ MPN/g, respectively. While the medians of heterotrophs at Daeam were larger than those at Sunam, the concentrations of naphthalene-degraders were higher at Sunam compared to those at Daeam. From the MPN tubes and enrichment cultures, we obtained 17 isolates of naphthalene-degraders which were identified as Sphingomonas paucimobilis, Brevundimonas vesic­ularis, Burkholderia cepacia, Ralstonia pickettii, Pseudomanas fluorescens, and Chryseomonas luteola. Among them, 6 isolates showed higher naphthalene-degrading activity on minimal media of pH 4 compared to pH 7, whereas the extent of growth was not greater at pH 4 than at pH 7 when they were inoculated on nutrient-rich media. It is plausible that the pH may affect naphthalene-degrading activity of the isolates by changing fatty acid composition of bacterial membrane.

Antifungal Property of Microorganisms against Korea Oak Wilt Pathogen, Raffaelea quercus-mongolicae (참나무시들음 병원균 Raffaelea quercus-mongolicae에 대한 항균미생물 분리)

  • Lee, Sang-Hyun;Lee, Seung-Kyu;Kim, Jae-Young;Lee, Chong-Kyu;Kim, Kyung-Hee;Yi, Yong-Sub
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.66-69
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    • 2012
  • Five strains out of 200 candidate strains (SG 1-9, 1-12, SG 2-8, 2-10, and 2-17) were selected to determine their antifungal activity against Raffaelea quercus-mongolicae. The 16S rDNA sequences of the five strains were determined by sequencing analysis and analyzed by the homology of the blast program at NCBI. The homology search showed that SG 1-9 and 1-12 had a 98% homology with Streptomyces cinnamoneus and 98% homology with Burkholderia cepacia, while SG 2-8, 2-10, and 2-17 had a 99% homology with Streptomyces fradiae, a 97% homology with Staphylococcus epidermidis, and a 99% homology with Staphylococcus epidermidis. Out of the five selected strains, organic extract and protein extracts of SG2-17 strain broth were employed to determine antifungal activity against Raffaelea quercus-mongolicae. The organic extract exhibited antifungal activity, but the protein extracts did not demonstrate such an activity. Three organic solvents, butanol, benzene, and ethyl acetate, were also used for determination of antifungal activities. The activity measurements revealed that benzene extract possessed the greatest inhibitory effect on the growth of Raffaelea quercus-mongolicae, with the next highest being butanol extract, and ethyl acetate extract being the lowest.

Development of Functional Halogenated Phenylpyrrole Derivatives (기능성 할로겐화 페닐피롤 )

  • Min-Hee Jung;Hee Jeong Kong;Young-Ok Kim;Jin-Ho Lee
    • Journal of Life Science
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    • v.33 no.10
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    • pp.842-850
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    • 2023
  • Pyrrolnitrin, pyrrolomycin, and pyoluteorin are functional halogenated phenylpyrrole derivatives (HPDs) derived from microorganisms with diverse antimicrobial activities. Pyrrolnitrin is a secondary metabolite produced from L-tryptophan through four-step reactions in Pseudomonas fluorescens, Burkholderia cepacia, Serratia plymuthica, etc. It is currently used for the treatment of superficial dermatophytic fungal infections, has high antagonistic activities against soil-borne and foliar fungal infections, and has many industrial applications. Since pyrrolnitrin is easily decomposed by light, it is difficult to widely use it outdoors. As an alternative, fludioxonil, a synthetically produced non-systemic surface fungicide that is structurally similar and has excellent light stability, has been commercialized for seed and foliar treatment of plants. However, due to its high toxicity to aquatic organisms and adverse effects in human cell lines, many countries have established maximum residue levels and strictly control its levels. Pyrrolomycin and pyoluteorin, which have antibiotic/antibiofilm activity against Gram-positive bacteria and high anti-oomycete activity against the plant pathogen Pythium ultimum, respectively, were isolated and identified from microorganisms. This review summarizes the biosynthesis and production of natural pyrrolnitrin derived from bacteria and the characteristics of synthetic fludioxonil and other natural phenylpyrrole derivatives among the HPDs. We expect that a plethora of highly effective, novel HPDs that are safe for humans and environments will be developed through the generation of an HPD library by microbial biosynthesis and chemical synthesis.

Analysis of Plasmid pJP4 Horizontal Transfer and Its Impact on Bacterial Community Structure in Natural Soil

  • KIM TAE SUNG;KIM MI SOON;JUNG MEE KUM;JOE MIN JEONG;AHN JAE HYUNG;OH KYOUNG HEE;LEE MIN HYO;KIM MIN KYUN;KA JONG OK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.376-383
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    • 2005
  • Alcaligenes sp. JMP228 carrying 2,4­dichlorophenoxyacetic acid (2,4-D) degradative plasmid pJP4 was inoculated into natural soil, and transfer of the plasmid pJP4 to indigenous soil bacteria was investigated with and without 2,4-D amendment. Plasmid pJP4 transfer was enhanced in the soils treated with 2,4-D, compared to the soils not amended with 2,4-D. Several different transconjugants were isolated from the soils treated with 2,4-D, while no indigenous transconjugants were obtained from the unamended soils. Inoculation of the soils with both the donor Alcaligenes sp. JMP228/pJP4 and a recipient Burkholderia cepacia DBO 1 produced less diverse transconjugants than the soils inoculated with the donor alone. Repetitive extragenic palindromic-polymerase chain reaction (REP-PCR) analysis of the transconjugants exhibited seven distinct genomic DNA fingerprints. Analysis of 16S rDNA sequences indicated that the transconjugants were related to members of the genera Burkholderia and Pandoraea. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that inoculation of the donor caused clear changes in the bacterial community structure of the 2,4-D­amended soils. The new 16S rRNA gene bands in the DGGE profile corresponded with the 16S rRNA genes of 2,4-D­degrading transconjugants isolated from the soil. The results indicate that introduction of the 2,4-D degradative plasmid as Alcaligenes sp. JMP228/pJP4 has a substantial impact on the bacterial community structure in the 2,4-D-amended soil.

가스상 TCE 처리를 위한 추출막 생물반응기의 수학적 모사

  • Kim, Ji-Seok;Kim, Gwan-Su;Jang, Deok-Jin
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.370-373
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    • 2000
  • In this work, an extractive membrane bioreactor containing coulture broth of Burkholderia cepacia G4 PR1 constitutively expressing the TCE-degrading enzyme, tolune-ortho-monooxygenase(TOM), was used for the degradation of TCE. The membrane bioreactor operates by seperating the TCE-containing waste gas from the aerated biomedium, by which the air-stripping of TCE without degradation was overcome that could occur in conventional aerobic biological treatments of TCE-contaminated waste gases. This was achieved by a silicone rubber membrane which was coiled around a perspex draft tube. TCE from the gas phase diffuses across the silicone rubber membrane into microbial culture broth that was continuously fed from a separate aerobic CSTR. Therefore, TCE degradation occured without the TCE being directly exposed to the aerating gas stream. Of the TCE supplied to the membrane bioreactor, 72.6% was biodegraded during the operation of this system. To construct a mathematical model for this system, parameters describing microbial growth kinetics on TCE were determined using a CSTR bioreactor. Else parameters used for numerical simulation were determined from either indepedent experiments or values reported in the literature. The model was compared with the experimental data, and there was a good agreement between the predicted and the measured TCE concentrations in the system. To achieve a higher treatment efficiency, various operating conditions were simulated as well.

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Utilization of biofilter/photo-catalytic reactor system for the simultaneous treatment of hydrogen sulfide and toluene from waste-air

  • Lim, Kwang-Hee;Ping, Wang Zhi;Lim, Dong-Joon;Lee, Eun-Ju
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.287-292
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    • 2005
  • Transient behavior of biofilter/photo-catalytic reactor system was observed to eliminate both hydrogen sulfide and toluene from waste air at its four sampling ports. The biofilter was packed with a equivolume mixture of granular activated carbon(GAC) and compost as packing media on which Thiobacillus sp. IW and Burkholderia cepacia G4 were inoculated and were fixed. The biofilter/photo-catalytic reactor system was run for eight stages of operation under various operating conditions. As a result the removal efficiencies of hydrogen sulfide and toluene began to decrease from 100% after the inlet loads of hydrogen sulfide and toluene surpassed ca. 100 $S-g/m^{3}/h$ and $161g/m^{3}/h$, respectively, and were rapidly decreased to 60% after the inlet loads of hydrogen sulfide and toluene were increased to 200 $S-g/m^{3}/h$ and $644g/m^{3}/h$, respectively.

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Effect of Electrochemical Oxidation Potential on Biofilter for Bacteriological Oxidation of VOCs to $CO_2$

  • Kang Hye-Sun;Lee Jong-Kwang;Kim Moo-Hoon;Park Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.399-407
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    • 2006
  • In this study, an electrical conductive carbon fiber was used as a biofilter matrix to electrochemically improve the biofilter function. A bioreactor system was composed of carbon fiber (anode), titanium ring, porcelain ring, inorganic nutrient reservoir, and VOC reservoir. Electric DC power of 1.5 volt was charged to the carbon fiber anode (CFA) to induce the electrochemical oxidation potential on the biofilter matrix, but not to the carbon fiber (CF). We tested the effects of electrochemical oxidation potential charged to the CFA on the biofilm structure, the bacterial growth, and the activity for metabolic oxidation of VOCs to $CO_2$, According to the SEM image, the biofilm structure developed in the CFA appeared to be greatly different from that in the CF. The bacterial growth, VOCs degradation, and metabolic oxidation of VOCs to $CO_2$ in the CFA were more activated than those in the CF. On the basis of these results, we propose that the biofilm structure can be improved, and the bacterial growth and the bacterial oxidation activity of VOCs can be activated by the electrochemical oxidation potential charged to a biofilter matrix.