• Title/Summary/Keyword: Pseudomonas sp. P20

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미생물을 이용한 원유 및 원유제품의 분해 특성

  • O, Gyeong-Taek;Park, Gwi-Hwan;Lee, Jeong-Il;Lee, Jung-Gi;Kim, Seong-Jun;Motoki, Kubo;Jeong, Seon-Yong
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.435-438
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    • 2000
  • Crude oil-degrading microorganisms, Acinetobacter sp. A132, Pseudomonas aeruginosa F722, and Acinetobacter calcoaceticus OM1 were isolated from soil and sea. The optimal temperature of strain A132 and strain F722 on growth isolated from soil was $35^{\circ}C$ both, and also their growth were optimized at pH 8 and 9, respectively. The growth of the strains, A132 and F722, showed that crude oil of 2% (w/v) in culture broth in which crude oil was used as carbon and energy sources appeared to be an optimum. Optimal culture conditions of strain OM1 were different from those of the soil microorganisms except for temperature. The growth of strain OM1 was optimized at pH 7 and crude oil of 3.0% (w/v). The degradability to crude oil by strain A132 showed maximum $5.49g/\;l\;{\cdot}\;day$ under the conditions of $25^{\circ}C$, NaCl of 1.0% (w/v), and crude oil of 2.0% (w/v). The highest degradability of strain F722 to crude oil was $1.19g/\;l\;{\cdot}\;day$ under the culture conditions at $35^{\circ}C$, NaCl 1.0% (w/v), and crude oil of 2.0% (w/v). The degradation characteristics of kerosene $(nC_9-nC_{20})$ and diesel $(nC_9-nC_{28})$ by strain OM1, and F722 were analyzed by gas chromatography. Strain OM1 degraded more than 95% of kerosene and 75% of diesel for 7 days cultivation. Strain F722 showed degradation of more than 80% to kerosene in 10 days.

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Purification and Characterization of Bacteriocin Produced by Enterococcus sp. (Enterococcus sp.가 생산하는 Bacteriocin의 정제 및 특성에 관한 연구)

  • 정건섭;양은석;이국진;고현정;정병문
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.523-528
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    • 1998
  • We isolated microorganism secreting antimicrobial substance from tomato and identified as Enterococcus faecium. This substance was completely inactivated by pretense treatment and retained activity after catalase treatment. This result indicated that the antimicrobial activity of this substance was due to proteinaceous substance known as bacteriocin. The bacteriocin inhibited growth of Gram positive bacteria, such as Listeria monocytogenes, Leuconostoc mesenteroides, Lactobacillus plantarum, Streptococcus agalactiae, Streptococcus pyrogenes, and Gram negative bacteria, such as Pseudomonas aeruginosa. Purification of the bacteriocin was achieved by ethanol precipitation, ion exchange chromatography on CM Sepharose CL-6B, and gel filtration on Sephacryl S-100 HR. After these purification steps, the specific activity of the bacteriocin was increased 35.8 fold compared with culture broth. Purified bacteriocin was shown single band on SDS-PAGE and molecular weight was estimated 51 kDa. The residual activity of this bacteriocin was 3.3% at 10$0^{\circ}C$ for 60 min, and this bacteriocin was stable at pH 2~7.

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Biosynthesis of Lactate-containing Polyhydroxyalkanoates in Recombinant Escherichia coli by Employing New CoA Transferases (재조합 대장균에서 새로운 코엔자임 에이 트랜스퍼레이즈를 이용한 젖산을 모노머로 함유한 폴리하이드록시알칸산 생산 연구)

  • Kim, You Jin;Chae, Cheol Gi;Kang, Kyoung Hee;Oh, Young Hoon;Joo, Jeong Chan;Song, Bong Keun;Lee, Sang Yup;Park, Si Jae
    • KSBB Journal
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    • v.31 no.1
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    • pp.27-32
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    • 2016
  • Several CoA transferases from Clostridium beijerinckii, C. perfringens and Klebsiella pneumoniae were examined for biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) in recombinant Escherichia coli XL1-Blue strain. The CB3819 gene and the CB4543 gene from C. beijerinckii, the pct gene from C. perfringens and the pct gene from K. pneumoniae, which encodes putative CoA transferase gene, respectively, was co-expressed with the Pseudomonas sp. MBEL 6-19 phaC1437 gene encoding engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 ($PhaC1_{Ps6-19}$) to examine its activity for the construction of key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli XL1-Blue expressing the phaC1437 gene and CB3819 gene synthesized poly(3-hydroxybutyrate) [P(3HB)] homopolymer to the P(3HB) content of 60.5 wt% when it was cultured in a chemically defined medium containing 20 g/L of glucose and 2 g/L of sodium 3-hydroxybutyrate. Expression of the phaC1437 gene and CB4543 gene in recombinant E. coli XL1-Blue also produced P(3HB) homopolymer to the P(3HB) content of 51.2 wt% in the same culture condition. Expression of the phaC1437 gene and the K. pneumoniae pct gene in recombinant E. coli XL1-Blue could not result in the production of PHAs in the same culture condition. However, the recombinant E. coli XL1-Blue expressing the phaC1437 gene and the C. perfringens gene could produce poly(3-hydroxybutyrate-co-lactate [P(86.4mol%3HB-co-13.7 mol%LA) up to the PHA content of 10.6 wt% in the same culture condition. Newly examined CoA transfereases in this study may be useful for the construction of engineered E. coli strains to produce PHA containing novel monomer such lactate.

Characterization of Diesel Oil-Degrading Bacteria (디젤유 분해균주의 특성 및 토양배양)

  • 안민정;한윤전;임현섭;최기현;권오범;정병철
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.108-113
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    • 2003
  • Diesel oil-degrading bacterial strains were isolated from diesel oil contaminated soil and called HS series (HS1, HS2 and HS3). These strains were identified as Acinetobacter sp. (HS1) and Pseudomonas sp. (HS2 and HS3) based on Biolog test, cellular fatty acid composition, and 16S rDNA sequence analysis. These strains were coltivated in liquid minimal media containing 2% diesel oil, and diesel oil-degrading activity was measured. As result, all strains degraded over 70% of total diesel oil. But PAH (polycyclic aromatic hydrocarbon)- and pris- tane-degrading rate of these strain was below 20% of total PAH and pristane. The HS 1 strain showed highest hydrophobicity and low emulsifying activity among the experimental strains and high diesel oil-degrading activity. From the above-mentioned result, microcosm experiment was performed with the HS1 strain. The HS1 strain showed a degrading activity of over 80% of total diesel oil in microcosm test. And microbial activity was correlated to diesel oil-degrading activity. Therefore, it is suggested that the HS1 strains could be effectively used for the bioremediation for diesel oil.

Biosynthesis of Lactate-containing Polyhydroxyalkanoates in Recombinant Escherichia coli from Sucrose (재조합 대장균에서 수크로즈로부터의 젖산을 모노머로 함유한 폴리하이드록시알칸산 생산 연구)

  • Oh, Young Hoon;Kang, Kyoung-Hee;Shin, Jihoon;Song, Bong Keun;Lee, Seung Hwan;Lee, Sang Yup;Park, Si Jae
    • KSBB Journal
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    • v.29 no.6
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    • pp.443-447
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    • 2014
  • Biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) was examined in recombinant Escherichia coli W strain from sucrose. The Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene, which encode engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 ($PhaC1_{Ps6-19}$) and engineered C. propionicum propionyl-CoA transferase ($Pct_{Cp}$), respectively, were expressed in E. coli W to construct key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli W expressing the phaC1437 gene and the pct540 gene could synthesize P(3HB-co-13mol%LA) up to the polymer content of 31.3 wt% when it was cultured in chemically defined MR medium containing 20 g/L of sucrose and 2 g/L of sodium 3-hydroxybutyrate. When Ralstonia eutropha phaAB genes were additionally expressed to provide 3-hydroxybutyrate-CoA (3HB-CoA) from sucrose, P(3HB-co-16mol%LA) could be synthesized from sucrose as a sole carbon source without supplement of sodium 3-hydroxybutyrate in culture medium, but the PHA content was decreased to 12.2 wt%. The molecular weight of P(3HB-co-16 mol%LA) synthesized in E. coli W using sucrose as carbon source were $1.53{\times}10^4$ ($M_n$) and $2.78{\times}10^4$ ($M_w$), respectively, which are not different from those that have previously been reported by other recombinant E. coli strains. Engineered E. coli strains developed in this study should be useful for the production of lactate-containing PHAs from sucrose, one of the most abundant and least expensive carbon sources.

Biochemical and Molecular Identification of Antibacterial Lactic Acid Bacteria Isolated from Kimchi (김치에서 항균활성 유산균의 분리 및 동정)

  • Kim, Soo-Young;Kim, Jong-Doo;Son, Ji-Soo;Lee, Si-Kyung;Park, Kab-Joo;Park, Myeong-Soo
    • Korean Journal of Food Science and Technology
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    • v.43 no.4
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    • pp.446-452
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    • 2011
  • Total 480 lactic acid-producing bacteria were isolated from five kinds of kimchi, and their antibacterial activity was tested against Salmonella enterica serovar Typhimurium, Bacillus subtilis, and Pseudomonas aeruginosa using an agar diffusion assay. Among them, 340 isolates showed a halo on MRS agar against one or more indicator strains, which were identified using multiplex PCR, an API 50CHL kit, and a 16S rDNA sequence analysis. As a result, 169 Lactobacillus plantarum, 20 Lactobacillus fermentum, two Lactobacillus paracasei ssp. paracasei, two Lactobacillus sp., and 15 Pediococcus sp. were identified. This may be the first report on the isolation of antibacterial Lactobacillus fermentum from kimchi.

Response of Microbe to Chemical Properties from Orchard Soil in Gyeongnam Province (경남지역 과수원 토양 화학성분이 미생물 생태에 미치는 영향)

  • Lee, Young-Han;Zhang, Yong-Sun
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.2
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    • pp.236-241
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    • 2011
  • Soil microbial diversity was responsible for a strong effect on the chemical properties of orchard soils. This study evaluated a relationship between soil chemical properties and soil microbial diversities at 25 sites in orchard soils in Gyeongnam Province. The average nutrients in the orchard soils were 2.6 times for available phosphorous, 2.3 times for exchangeable potassium and 1.3 times for exchangeable calcium higher compared to recommend concentrations in the orchard soils. Contents of available phosphorous and organic matter in the inclined piedmont soils were higher than those in the other topographical soils (p<0.05). Populations of fungi and fluorescence Pseudomonas sp. in the silt loam soils were significantly higher than those in the sandy loam soils (p<0.05). In principal component analysis of chemical properties and microbial populations in the upland soils, our findings suggested that population of bacteria should be considered as potential factor responsible for the clear orchard soils differentiation. The soil organic matter was significantly negative correlation with population of bacteria whereas was positive correlation with population of fungi in orchard soils.

Studies on the promoted Aging of flue-cured leaf tobacco by cellulolytic Enzyme and Nicotinophiles (섬유소 분해효소 및 니코틴 분해세균을 이용한 잎담배의 발효촉진효과)

  • 이태호;성낙계
    • Journal of the Korean Society of Tobacco Science
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    • v.13 no.2
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    • pp.5-20
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    • 1991
  • For the quality enhancement of harvested-year leaf tobacco to the quality of 2-year naturally aged leaf tobacco, cellulose and nicotine degradative bacteria were isolated and identified. Effects of artificial fermentation treated cellulase and nicotine degradative bacteria on the quality of leaf tobacco were investigated from the chemical and sensory points of view. 1, Changes in chemical composition of leaf tobacco resulted from the addition of cellulase extracted from Cellulomonas sp. [3ml(${\mu}{\textrm}{m}$ D-glucose/ml. mil-1) of enzymes solution 11009 of leaf tobacco] and nicotine degradative bacteria, Pseudomonas sp. 2ml(IX109 cells$\div$ 100g of leaf tobacco), and subsequently fermented at 40${\mu}{\textrm}{m}$$^{\circ}C$, 65% R. H. for 40 days are as follows : 1) Content of crude fiber decreased 12% It took 9 min, 53 sec. to reach full combustion in control group but took only 7 min. 47 sec. in the treated group, taking almost equal time to 2-year naturally aged leaf tobacco(7 min. 35sec.). 2) Light intensity of control group was 60.96% with bright lemon color but that of treated leaf tobacco accounted for 47.69 with orange to dark brown color series, which was almost equal to the value, 45.69, of 2-year naturally aged leaf tobacco. 3) Linoleic acid, serving mild taste among organic acids, amounted to 1.llmg/g in control group but increased to 1.35m9/9 in the treated leaf tobacco, identical to the content(1.35mg/g) of 2-year naturally aged leaf tobacco. 4) Content of solanone, on of the typical leaf tobacco flavor compounds, accounted for 2.95% in control group but increased to 2.87% in treated group. 5) Methyl furan, useful flavor compound in smoke composition, accounted for 17.6$\mu\textrm{g}$/cig. in control group but increased to 25.9$\mu\textrm{g}$/cig. in treated group. However, acroleine decreased from 69.3$\mu\textrm{g}$/cig. in control group to 58.6$\mu\textrm{g}$/cig. in treated group 2. In sonsory test, mild taste evaluation of control group scored 5.47 and treated group 7.93 which was evaluted almost equal to the value(8.00) of 2-year naturally aged leaf tobacco. Aroma evaluation of control group scored 5.60, treated group 8.20, and 2-year naturally aged leaf tobacco 8.33. In addition, total harmony taste of control group showed 5.67, treated group 8.07 (p<0.01), and 2-year naturally aged leaf tobacco 8.00. From these results, it can be said that quality of treated leaf tobacco is not inferior to that 2-year naturally aged leaf tobacco.

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Changes in Physicochemical Properties and Microbial Population during Fermenting Process of Organic Fertilizer (혼합발효 유기질비료의 발효과정 중 이화학성 및 미생물밀도 변화)

  • Lee, Jong-Tae;Lee, Chan-Jung;Kim, Hee-Dae
    • Korean Journal of Soil Science and Fertilizer
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    • v.37 no.2
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    • pp.116-123
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    • 2004
  • This study was conducted to investigate the changes in physicochemical and microbiological properties during fermenting process of organic fertilizer which was made from the mixture of organic materials such as sesame oil cake, fish meal, blood meal, rice bran, ground bone meal, and natural minerals such as illite, crusted oyster shell and loess. They were mixed and fermented for 70 days. The sesame oil cake and rice bran, major ingredients for organic fertilizers, consisted of 7.6 and 2.6% total nitrogen, 3.6 and 4.6% $P_2O_5$, 1.4 and 2.2% $K_2O$, respectively. The ground bone meal included 29.2% $P_2O_5$ and illite included 3.8% $K_2O$. Temperature of organic fertilizer during the fermentation rapidly increased over $50^{\circ}C$ within 2 days after mixing and stabilized similar to outdoor temperature after 40 days. Moisture content decreased from 36.3 to 16.0% after 1 month. C/N ratio of organic fertilizer slightly increased until 30 days and thereafter, it slowly decreased, It resulted from the faster decrease of total nitrogen concentration compared with organic matter. Concentration of $NH_4-N$ in organic fertilizer rapidly increased from 1,504 to $5,530mg\;kg^{-1}$, the highest concentration after 10 days. Meantime, $NO_3-N$ concentration was low and constant about $150mg\;kg^{-1}$ over the whole fermenting period. This result seemed to be due to the high pH. The organic ferfilizer fermented for 70 days was composed of 2.7% N, 2.8% $P_2O_5$, 1.8% $K_2O$, and 35.9% organic matter. Total populations of aerobic bacteria, Bacillus sp. and actinomycetes, after fermenting process, were $12.5{\times}10^{10}$, $45.5{\times}10^{5}$ and $13.6{\times}10^{5}cfu\;g^{-1}$ respectively. Pseudomonas sp. was $71.9{\times}10^{7}cfu\;g^{-1}$ at first, but it rapidly decreased according to the rise of temperature. Yeasts played an important role in the early stage of fermentation and molds did in the late stage.

Biological Control of Phytophthora Blight of Red-pepper Caused by Phytophthora capsici.;II. Isolation and Antifungal Activity of the Substances (고추역병균(疫病菌)(Phytophthora capsici)의 생물학적(生物學的) 방제(防除);II. 항균물질(抗菌物質)의 분리(分離) 정제(精製) 및 항균활성(抗菌活性))

  • Chang, Yoon-Hee;Chang, Sang-Moon;Choi, Jyung;Lee, Dong-Hoon
    • Korean Journal of Environmental Agriculture
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    • v.15 no.4
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    • pp.399-405
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    • 1996
  • In the culture medium, the three antifungal fractions against P. capsici were separated by Sephadex G-25 column chromatography and Silica-gel chromatography. The substance A in white powder and the substance B in sticky oil were isolated by ethyl acetate : acetone mixture(7 : 3), and the substance C in yellow powder was isolated by chloroform : ethyl acetate mixture(95 : 5). The crude extract by ethyl acetate from the culture medium acidified to pH 2 was known to inhibit completely the growth of P. capsici at the level of $50mgkg^{-1}$. The substance A and B were known to be effective above the level of $5mgkg^{-1}$, and the substance C was effective above the level of $1mgkg^{-1}$. However, at the level of $20mgkg^{-1}$, the efficiency was in the order of A>C>B. It is apparent on a pot-experiment scale that the three substances effectively control Phytophthora blight of the red-pepper plant grown in the soil inoculated with P. capsici.

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