• Title/Summary/Keyword: Carbon Source

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Structure Analysis of pmcABCDEFT Gene Cluster for Degradation of Protocatechuate from Comamonas sp. Strain DJ-12 (Comamonas sp. Strain DJ-12로부터 Protocatechuate의 분해에 관여하는 pmcABCDEFT 유전자군의 구조 분석)

  • Kang Cheol-Hee;Lee Sang-Mhan;Lee Kyoung;Lee Dong-Hun;Kim Chi-Kyung
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.195-200
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    • 2005
  • Comamonas sp. strain DJ-12 is a bacterial isolate capable of degrading of 4-chlorobiphenyl (4CB) as a carbon and energy source. The degradation pathway was characterized as being conducted by consecutive reactions of the meta-degradation of 4CB, hydrolytic dechlorination of 4-chlorobenzoate (4CBA), hydroxylation of 4-hydroxybenzoate, and meta-degradation of protocatechuate to product TCA metabolites. The 6.8 kb fragment from the chromosomal DNA of Comamonas sp. strain DJ-12 included the genes encoding for the meta-degradation of PCA; the genes of protocatechuate 4,5-dioxygenase alpha and beta subunits (pmcA and pmcB), 4-carboxy-2-hydroxymuconate-6-semialdehyde dehydrogenase (pmcC), 2-pyrone-4,6-dicarboxylate hydrolase (pmcD), 4-oxalomesaconate (OMA) hydratase(pmcE), 4-oxalocitramalate (OCM) aldolase (pmcF), and transporter gene (pmcT). They were organized in the order of pmcT-pmcE-pmcF-pmcD-pmcA-pmcB-pmcC. The amino acid sequences deduced from the nucleotide sequences of pmcABCDEFT genes from Comamonas sp. strain DJ-12 exhibited 94 to $98\%$ homologies with those of Comamonas testosteroni BR6020 and Pseudomonas ochraceae NGJ1, but only 52 to $74\%$ with homologies Sphingomonas paucimobilis SYK-6, Sphingomonas sp. LB126, and Arthrobacter keyseri 12B.

Isolation and Characterization of Bacteria Capable of Degrading Bisphenol A (Bisphenol A 분해세균의 분리 및 특성)

  • 김희식;이영기;이완석;박찬선;윤병대;오희목
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.189-196
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    • 2001
  • Eighty-seven microbial strains capable of growing on bisphenol A (BPA) as a sole carbon source were isolated from soils, waste waters and sludges. Among them, three bacterial strains were finally selected as potential decomposers through measuring BPA-degradation efficiency by HPLC analysis. Two of these bacterial strains were identified as Serratia marcescens 1901 and S. marcescens 1902, and another was Pseudomonas putida 1401 by 16S rDNA partial sequences and based on morphological and physiological properties. They showed higher cell growth and BPA degradation in PAV (PAS medium containing vitamin mixtures) than in PAS medium. The degradation efficiencies of these bacterial strains were within a range of 20-40% in the PAV containing 500 mg/1 or 100 mg/l of BPA fur 3 days. S. marcescens 1901 showed higher degradation efficiency at 100 mg/1 of BPA than those of other selected strains, while S. marcescens 1902 and P. putida 1401 degraded a high concentration of BPA (500 mg/l) with a degradation efficiency of 40% for 3 days. The BPA degradation using a mixed culture of three selected strains showed the similar level of dog-radation efficiency with that using a pure culture.

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The Uptake of 2-deoxy-D-glucose (2dGlc) by the Endogenous Sugar Transporter(s) of Spodoptera frugiperda Clone 21-AE Cells and the Inhibition of 2dGIc Transport in the Insect Cells by Fructose and Cytoc halasin B

  • Lee, Chong-Kee
    • Biomedical Science Letters
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    • v.9 no.4
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    • pp.177-181
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    • 2003
  • The baculovirus/Spodoptera frugiperda (Sf) cell system has become popular for the production of large amounts of the human erythrocyte glucose transporter, GLUT1, heterologously. However, it was not possible to show that the expressed transporter in insect cells could actually transport glucose. The possible reason for this was that the activity of the endogenous insect glucose transporter was extremely high and so rendered transport activity resulting from the expression of exogenous transporter very difficult to detect. Sf21-AE cells are commonly employed as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains 0.1 % D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike the human glucose transporter, very little is known about properties of the endogenous sugar transporter(s) in insect cells. Thus, the uptake of 2-deoxy-D-glucose (2dGlc) by Sf21-AE cells and the inhibition of 2dGlc transport in the insect cells by fructose and cytochalasin B were investigated in the present work. The binding assay of cytochalasin B was also performed, which could be used as a functional assay for the endogenous glucose transporter(s) in the insect cells. Sf21-AE cells were infected with the recombinant virus AcNPV-GT or no virus, at a multiplicity of infection (MOI) of 5. Infected cells were resuspended in PBS plus and minus 300 mM fructose, and plus and minus 20 $\mu$M cytochalasin B for use in transport assays. Uptake was measured at 28$^{\circ}C$ for 1 min, with final concentration of 1 mM deoxy-D-glucose, 2-[1,2-$^3$H]- or glucose, L-[l,$^3$H]-, used at a specific radioactivity of 4 Ci/mol. The results obtained demonstrated that the sugar uptake in uninfected cells was stereospecific, and was strongly inhibited by fructose but only poorly inhibitable by cytochalasin B. It is therefore suggested that the Sf21-AE glucose transporter has very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.

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Effects of Nutrient Source on Soil Physical, Chemical, and Microbial Properties in an Organic Pear Orchard (유기질 비료 급원이 배 과원의 토양 물리화학성 및 미생물성에 미치는 영향)

  • Choi, Hyun-Sug;Li, Xiong;Kim, Wol-Soo;Lee, Youn
    • Korean Journal of Environmental Agriculture
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    • v.30 no.1
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    • pp.16-23
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    • 2011
  • BACKGROUND: This study was conducted to investigate the effects of different organic treatments and a chemical fertilizer on the soil chemical, physical, and microbial properties in an organic pear orchard. METHODS AND RESULTS: Control was referred as a NPK chemical fertilizer (15N-9P-10K) and organic treatments included compost containing with oil cake, compost containing with humic acid, and compost containing with chitin substance. All treatments applied at rates equivalent to 200 g N per tree per year under the tree canopy in March 30 of 2008 and 2009. Soil bulk density, solid phase, liquid phase, and penetration resistance were not significantly different among the treatments. Organic treatment plots had greater organic matter, total nitrogen, potassium, and magnesium concentrations compared to control, and the nutrient concentrations were not consistently affected by the organic treatments. Microbial biomass nitrogen and carbon, dehydrogenase, acid-phosphatase, and chitinase activities overall increased from March to August. Organic treatments, especially compost containing with oil cake or chitin aicd, increased the microbial variables compared to control. CONCLUSION(s): All the organic treatments consistently stimulated soil biological activity. The consistent treatment effect, however, did not occur on the soil mineral nutrition as the trees actively taken up the nutrients during a growing season, which would have diminished treatment effects. Long-term study required for evaluating soil physical properties in a pear orchard.

Phosphorus Removal (Characteristics by Anoxic Oxic Process) by Anoxic and Oxic Processed Combined with Iron Electrolysis (철 석출장치가 결합된 무산소.호기공정에 의한 인 제거 특성)

  • Kim, Min-Ho;Kim, Young-Gyu;Kim, Soo-Bok
    • Journal of Environmental Health Sciences
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    • v.36 no.6
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    • pp.502-509
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    • 2010
  • In this study, the (phosphorous removal) the characteristics of phosphorous removal due to (the iron compound precipitated) iron compound precipitation by iron electrolysis in (the anoxic. oxic process) anoxic and oxic processes (equipped with the) in an iron precipitation device were analyzed. During the device operation period, the average concentration of BOD, T-N, and T-P were 219.9 mg/l, 54.6 mg/l and 6.71 mg/l, respectively. The BOD/$COD_{Cr}$ ratio was 0.74, and the BOD/T-N and BOD/T-P ratios were 4.0 and 32.8, respectively. The removal rate of (the organic matters) organic matter (BOD and $COD_{Cr}$) was very high at 91.6% or higher, and that of nitrogen was 80.5%. The phosphorous concentration (of the final) in the treated water was 0.43 mg/l (0.05-0.74 mg/l) on average, and the removal efficiency was high at 90.8%. The soluble T-P concentrations in (an) the anoxic reactor, oxic reactor (II) and final treated water were 1.99 mg/l, 0.79 mg/l and 0.43 mg/l, respectively, which indicated that the phosphorous concentration in the treated water was very low. Regardless of the changes in the concentrations of (organic matters) organic matter, nitrogen and phosphorous in the influent, the quality of the treated water was relatively stable and high. The removal rate of T-P somewhat increased with the increase in the F/M ratio in the influent, and it also linearly increased in proportion to the T-P loading rate in the influent. In the treatment process used in this study, phosphorous was removed (using) by the precipitated iron oxide. Therefore, the consumption of organic (matters) matter for biological phosphorus removal was minimized and (most of the organic matters were) was mostly used as the organic carbon source for the denitrification in the anoxic reactor. This (can be an economic) treatment process (without the need for the supply of additional organic matters) is economic and does not require the supply of additional organic matter.

Nitrogen, Phosphorus, and Organic Carbon Discharges in the Imgo Small Agricultural Watershed Catchment (임고천상류 소규모 농업유역에서 하천으로의 질소, 인 및 유기물의 부하)

  • Chung, Jong-Bae;Kim, Min-Kyeong;Kim, Bok-Jin;Park, Woo-Churl
    • Korean Journal of Environmental Agriculture
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    • v.18 no.1
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    • pp.70-76
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    • 1999
  • Since high concentrations of N, P, and organic C cause the excessive eutrophication in water systems, the control of nutrient export from agricultural nonpoint sources has become important. This study was conducted to estimate discharges of N, P, and organic C from a small agricultural watershed of the upper Imgo stream in Youngchun, Kyongbuk. Of the total area(1.420ha), 25% was agricultural land including paddy, upland and orchards and most of the remainder was forest. The resident population in the watershed was 194 in 80 households and relatively small numbers of livestocks including cow were raised. Mean concentrations of nutrients in the stream water were 4.95, 0.80, 6.72, 0.07 and 2.52mg/L for $NO_3-N$, $NH_4-N$, Total N, Total P and COD respectively. Annual discharges in 1997 were 28,991kg of $NO_3-N$. 3,010kg of $NH_4-N$, 37,006kg of Total N. 590kg of Total P, and 29,138kg of COD. There was a strong positive relationship between stream flow and precipitation, and also most of the nutrient discharges occurred in the rainy season (May to August). Since there was no any other industries in the watershed, agricultural practices and sewage from the resident households, forest runoff and livestock wastes were the major sources of NPS discharges. A combination of management options, including management of soil erosion and fertilizer application, could lead to reductions in nutrient exports.

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Effect of Medium Components and Culture Methods on Prothallus Propagation of Pteridium aquilinum var. latiusculum (Desv.) Underw. ex Hell. (고사리 전엽체의 증식에 미치는 배지구성물질과 배양방법의 영향)

  • Shin, So-Lim;Lee, Moo-Yeul;Choi, Jae-Sun;Lee, Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.22 no.4
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    • pp.337-342
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    • 2009
  • Present studies were conducted to evaluate the effects of medium strength(MS and Hyponex), carbon sources and their concentrations, agar concentrations, and inoculation amounts on prothallus propagation of Pterdium aquilinum var. latiusculum(Desv.) Underw. ex Hell in vitro. The optimum MS medium strength for prothallus propagation was 2MS concentration. Phosphate source was most effective for prothallus growth of P. aquilinum var. latisculum. The addition of 1% sucrose or glucose to MS medium promoted prothallus multiplication. Growth of prothallus was not affected by agar concentration. Propagation of homogenized prothallus was vigorous even in liquid medium. Chopped gametophytes(100 and 200 mg) were inoculated on 250 ml ${\Delta}$flask with 100 mL of 2MS concentration medium and suspension culture was done at 100 rpm for 22 days. After 20 days, prothallus multiplication slowed down, so 100 mg of chopped prothalli is recommended for initial inoculation, since initial amount of inoculum did not affect subsequent prothallus multiplication. Consequently after 20 days of suspension culture, prothallus should be subcultured or transplanted outside of growing vessels.

Studies on the Artificial Cultivation of Lentinus edodes on sawdust media (표고버섯 톱밥 인공재배(人工栽培)에 관(關)한 연구(硏究))

  • Kim, Han-Kyung;Park, Yong-Hwan;Cha, Dong-Yule;Chung, Hwan-Chae
    • The Korean Journal of Mycology
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    • v.15 no.1
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    • pp.42-47
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    • 1987
  • 4 strains of the oak mushroom Lentinus edodes including the strain ASI 3046 were collected home and abroad to investigate the characterics of mycelial growth on synthetic media and sawdust media. The possibility of artificial culture of Lentinus edodes on media composed of different saw dusts and adding materials was investigated. Results were as follows: The optimum temperature for growth ranged from $22^{\circ}C$ to $26^{\circ}C$ among the lineages of Lentinus edodes. The optimal growth temperature for ASI 3046 strain and ASI 3047 strain was $26^{\circ}C$ and $22^{\circ}C$, respectively. The optimal carbon source for ASI 3046 strain was glucose, and peptone and ammoinum tartrate were proved excellent as nitrogen sources. When Lentinus edodes was cultured on sawdust media by varying the mixing ratio of different sawdusts and adding materials, mixed sawdust media composed of oak sawdust(50%, v/v), poplar sawdust(50%, v/v), acorn powder(5%, v/v) and rice bran(10%, v/v) showed higher yields of fruit bodies. Results showed that the ideal strain of Lentinus edodes for artificial culture on sawdust media was strain ASI 3046.

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Characterization and N-Terminal Amino Acid Sequence Analysis of Catechol 2,3-dioxygenase Isolated from the Aniline Degrading Bacterium, Delftia sp. JK-2 (Aniline 분해세균 Delftia sp. JK-2에서 분리된 catechol 2,3-dioxygenase의 특성 및 N-말단 아미노산 서열분석)

  • 황선영;송승열;오계헌
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.1-7
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    • 2003
  • The aim of this work was to investigate the characterization and sequence of catechol 2,3-dioxygenase isolated from Delfia sp. JK-2, which could utilize aniline as sole carbon, nitrogen and energy source. In initial experiments, several characteristics of C2,3O separated with ammonium sulfate precipitation, DEAE-sepharose were investigated. Specific activity of C2,3O was approximately 4.72 unit/mg. C2,3O demonstrated its enzyme activity to other substrates, catechol and 4-methylcatechol. The optimum temperature of C2,3O was $$Cu^{2+}$^{\circ}C$, and the optimal pH was approximately 8. Metal ions such as $Ag^{+}$, $Hg^{+}$, and $Cu^{2+}$ showed inhibitory effect on the activity of C2,3O. Molecular weight of the enzyme was determined to approximately 35 kDa by SDS-PAGE. N-terminal amino acid sequence of C2,3O was analyzed as $^{1}MGVMRIG-HASLKVMDMDA- AVRHYENV^{26}$, and exhibited high sequence homology with that of C2,30 from Pseudomonas sp. AW-2, Comamonas sp. JS765, Comamonas testosteroni and Burkholderia sp. RPO07. PCR product was amplified with the primers derived from N-terminal amino acid sequence. In this work, we found that the amino acid sequence of Delftia sp. JK-2 showed high sequence homology of C2,3O from Pseudomonas sp. AW-2 (100%) and Comamonas sp. JS765 (97%).

Isolation and Characterization of xylR/TMutants in Escherichia coli (대장균(大腸菌)의 xylRjT 변이주(變異株)의 분리(分離) 및 그 특성(特性))

  • Roh, Dong Hyun;Rhee, In Koo
    • Current Research on Agriculture and Life Sciences
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    • v.10
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    • pp.125-135
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    • 1992
  • Nine strains of xyl mutants that could not utilize xylose as a carbon source were isolated from E. coli JM109 by the treatment of NTG in order to investigate the regulation of xylose operon and to use recipient cells for the cloning of xylR gene. For the characterization of all isolated mutants, colony colors of all mutants on MacConkey-xylose and MacConkey-xylulose agar plate were observed for the utilization of xylose and xylulose, and the growth level and the activity of xylose isomerase and xylulokinase were determined in need. The isolated xylR/T mutants formed the white colony on MacConkey-xy-lose and MacConkey-xylulose agar plate. They did not detect the activity of xylose isomerase, and the activity of xylose isomerase was not restored in transformants of xylR/T mutant with pEX13 which contained xylA gene. xylR and xylT mutants were classified from xylR/T mutants depending upon the growth level in minimal medium. xylT mutants; DH13, DH121 and DH125 could grow a little in that medium, but xylR mutants; DH10, DH53, and DH60 could not grow that medium.

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