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Construction of a Novel Shuttle Vector for Tetragenococcus species based on a Cryptic Plasmid from Tetragenococcus halophilus

  • Min Jae Kim;Tae Jin Kim;Yun Ji Kang;Ji Yeon Yoo;Jeong Hwan Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.211-218
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    • 2023
  • A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Emr), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

Characterization of Ni-YSZ cermet anode for SOFC prepared by glycine nitrate process (Glycine nitrate process에 의한 제조된 SOFC anode용 Ni-YSZ cermet의 물성)

  • Lee, Tae-Suk;Ko, Jung-Hoon;Lee, Kang-Sik;Kim, Bok-Hee
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.21 no.1
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    • pp.21-26
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    • 2011
  • Ni-YSZ (Yttria Stabilized Zirconia) composite powders were fabricated by glycine nitrate process. The prepared powders were sintered at $1300{\sim}1400^{\circ}C$ for 4 h in air and reduced at $1000^{\circ}C$ for 2 h in a nitrogen and hydrogen atmosphere. The microstructure, electrical conductivity, thermal expansion and mechanical properties of the Ni-YSZ cermets have been investigated with respect to the volume contents of Ni. A porous microstructure consisting of homogeneously distributed Ni and YSZ phases together with well-connected grains was observed. It was found that the open porosity, electrical conductivity, thermal expansion and bending strength of the cermets are sensitive to the volume content of Ni. The Ni-YSZ cermet containing 40 vol% Ni was ascertained to be the optimum composition. This composition offers sufficient open porosity of more than 30 %, superior electrical conductivities of 917.4 S/cm at $1000^{\circ}C$ and a moderate average thermal expansion coefficient of $12.6{\times}10^{-6}^{\circ}C^{-1}$ between room temperature and $1000^{\circ}C$.

BaCeO3-BaZrO3 Solid Solution (BCZY) as a High Performance Electrolyte of Protonic Ceramic Fuel Cells (PCFCs) (BaCeO3-BaZrO3 고용체(BCZY) 기반 프로톤 세라믹 연료전지(PCFC)용 고성능 전해질 개발)

  • An, Hyegsoon;Shin, Dongwook;Choi, Sung Min;Lee, Jong-Ho;Son, Ji-Won;Kim, Byung-Kook;Je, Hae June;Lee, Hae-Weon;Yoon, Kyung Joong
    • Journal of the Korean Ceramic Society
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    • v.51 no.4
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    • pp.271-277
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    • 2014
  • To overcome the limitations of the solid oxide fuel cells (SOFCs) due to the high temperature operation, there has been increasing interest in proton conducting fuel cells (PCFCs) for reduction of the operating temperature to the intermediate temperature range. In present work, the perovskite $BaCe_{0.85-x}Zr_xY_{0.15}O_{3-\delta}$ (BCZY, x = 0.1, 0.3, 0.5, and 0.7) were synthesized via solid state reaction (SSR) and adopted as an electrolyte materials for PCFCs. Powder characteristics were examined using X-ray diffraction (XRD), thermogravimetric analysis (TGA) and Brunauer, Emmett and Teller (BET) surface area analysis. Single phase BCZY were obtained in all compositions, and chemical stability was improved with increasing Zr content. Anode-supported cell with $Ni-BaCe_{0.55}Z_{0.3}Y_{0.15}O_{3-\delta}$ (BCZY3) anode, BCZY3 electrolyte and BCZY3-$Ba_{0.5}Sr_{0.5}Co_{0.8}Fe_{0.2}O_{3-\delta}$ (BSCF) composite cathode was fabricated and electrochemically characterized. Open-circuit voltage (OCV) was 1.05 V, and peak power density of 370 ($mW/cm^2$) was achieved at $650^{\circ}C$.

Studies on LiF-${Li_2}O-{B_2}{O_3}-{P_2}{O_5}$ based Glassy Solid Electrolytes (LiF-${Li_2}O-{B_2}{O_3}-{P_2}{O_5}$계 유리고체전해질에 관한 연구)

  • Park, Gang-Seok;Gang, Eun-Tae;Kim, Gi-Won;Han, Sang-Mok
    • Korean Journal of Materials Research
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    • v.3 no.6
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    • pp.614-623
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    • 1993
  • Electrical characteristics of LiF-$Li_{2}O-B_{2}O_{3}-P_{2}O_5$ glasses with fixed $Li_2O$ content have been investigated by using AC impedance spectroscopy. Part of the total lithium ions present in these glasses contributes to conduction, and the changes in electrical conductivity with composition was inconsistent with the weak electrolyte model. The power law could not be used to determine the hopping ion concentration in these glasses. Both mobile carrier density and mobility have been modified as Li were added in the form of LiF. The formation of $(B-O-P)^-,di^-$, and metaborate group gave additional available sites for Li+ diffusion causing the enhancement of conductivity. The observed maximum conductivity was $2.43 \times 10^{-4}$S/cm at $150^{\circ}C$ at the composition containing 8mol% LiF. The decomposion potential amounted to 5.94V. The Li/glass electrolyte/$TiS_2$ solid-state cell showed open circuit voltage of 3.14V and energy density of 22 Wh/Kg at $150^{\circ}C$.

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Production of Biodiesel and Nutrient Removal of Municipal Wastewater using a Small Scale Raceway Pond (미세조류 옥외 배양시스템을 이용한 바이오디젤 생산 및 도시하수 영양 염류 제거)

  • Kang, Zion;Kim, Byung-Hyuk;Oh, Hee-Mock;Kim, Hee-Sik
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.207-214
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    • 2013
  • A concerted effort to develop alternative forms of energy is underway due to fossil fuel shortages and its deleterious effects. Recently, bioenergy from microalgae has gained prominence and the use of municipal wastewater as a low cost alternative for a nutrient source has significant advantages. In this study, we have employed municipal wastewater directly after primary treatment (primary settling basin) in a small scale raceway pond (SSRP) for microalgal growth. Indigenous microalgae in the wastewater were encouraged to grow in the SSRP under optimal conditions. The mean removal efficiencies of TN, TP, and $NH_3-N$ after 6 days were 77.77%, 63.55%, and 89.02%, respectively. The average lipid content of the microalgae was 19.51% of dry cell weight, and linolenate and linoleate (18:n) were the predominant fatty acids. The 18S rRNA gene analysis and microscopic observations of the indigenous microalgae community revealed the presence of Chlorella vulgaris and Scenedesmus obliquus as the dominant microalgae. These results indicate that untreated municipal wastewater, serving as an excellent nitrogen and phosphate source for microalgal growth, could be treated using microalgae in open raceway ponds. Moreover, microalgal biomass could be further profitable by the extraction of biodiesel.

Characterization of a Small Cryptic Plasmid from Pseudomonas nitroreducens Strain TX1 (Pseudomonas nitroreducens TX1에 존재하는 작은 플라스미드의 특성 규명)

  • Nguyen, Ngoc Tuan;Lee, Kyoung;Kang, Ju Beom;Huang, Shir-Ly
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.210-215
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    • 2014
  • Pseudomonas nitroreducens TX1 was isolated from a rice field drainage in Taiwan. The bacterium is of special interest because of its capability to use a group of nonionic surfactants such as alkylphenol polyethoxylates even at high concentrations as a sole carbon source. In this study, a small cryptic circular plasmid, pTX1, was characterized from P. nitroreducens TX1. It is 2,286 bp in length with a GC content of 63.3% and harbors three open reading frames, $Rep_{pTX1}$ and functionally unidentified ORF1 and ORF2. The predicted $rep_{pTX1}$ gene product is homologous to Rep proteins of plasmids belonging to the pC194/pUB110 family, which is predominantly found in Gram-positive bacteria and is known to replicate by the rolling-circle mechanism. The copy number of pTX1 was estimated to be about 150 in each cell. Based on the genetic fingerprints and comparison with other plasmids, it is concluded that pTX1 replicates by a rolling circle mechanism which is rarely found for Pseudomonas plasmids.

Confirmation of Drought Tolerance of Ectopically Expressed AtABF3 Gene in Soybean

  • Kim, Hye Jeong;Cho, Hyun Suk;Pak, Jung Hun;Kwon, Tackmin;Lee, Jai-Heon;Kim, Doh-Hoon;Lee, Dong Hee;Kim, Chang-Gi;Chung, Young-Soo
    • Molecules and Cells
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    • v.41 no.5
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    • pp.413-422
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    • 2018
  • Soybean transgenic plants with ectopically expressed AtABF3 were produced by Agrobacterium-mediated transformation and investigated the effects of AtABF3 expression on drought and salt tolerance. Stable Agrobacterium-mediated soybean transformation was carried based on the half-seed method (Paz et al. 2006). The integration of the transgene was confirmed from the genomic DNA of transformed soybean plants using PCR and the copy number of transgene was determined by Southern blotting using leaf samples from $T_2$ seedlings. In addition to genomic integration, the expression of the transgenes was analyzed by RT-PCR and most of the transgenic lines expressed the transgenes introduced. The chosen two transgenic lines (line #2 and #9) for further experiment showed the substantial drought stress tolerance by surviving even at the end of the 20-day of drought treatment. And the positive relationship between the levels of AtABF3 gene expression and drought-tolerance was confirmed by qRT-PCR and drought tolerance test. The stronger drought tolerance of transgenic lines seemed to be resulted from physiological changes. Transgenic lines #2 and #9 showed ion leakage at a significantly lower level (P < 0.01) than ${\underline{n}}on-{\underline{t}}ransgenic$ (NT) control. In addition, the chlorophyll contents of the leaves of transgenic lines were significantly higher (P < 0.01). The results indicated that their enhanced drought tolerance was due to the prevention of cell membrane damage and maintenance of chlorophyll content. Water loss by transpiration also slowly proceeded in transgenic plants. In microscopic observation, higher stomata closure was confirmed in transgenic lines. Especially, line #9 had 56% of completely closed stomata whereas only 16% were completely open. In subsequent salt tolerance test, the apparently enhanced salt tolerance of transgenic lines was measured in ion leakage rate and chlorophyll contents. Finally, the agronomic characteristics of ectopically expressed AtABF3 transgenic plants ($T_2$) compared to NT plants under regular watering (every 4 days) or low rate of watering condition (every 10 days) was investigated. When watered regularly, the plant height of drought-tolerant line (#9) was shorter than NT plants. However, under the drought condition, total seed weight of line #9 was significantly higher than in NT plants (P < 0.01). Moreover, the pods of NT plants showed severe withering, and most of the pods failed to set normal seeds. All the evidences in the study clearly suggested that overexpression of the AtABF3 gene conferred drought and salt tolerance in major crop soybean, especially under the growth condition of low watering.

Effects of Sprinkler and Fan Cooling System in Loose Barn on the Physiological Parameter and Milk Productivity in Holstein Cows During Summer (개방식우사내 Fan과 Sprinkler설치가 착유우의 생리적 변화 및 유생산성에 미치는 영향)

  • 정태영;이현민;김종민;김동일;이연섭;이인형
    • Journal of Animal Environmental Science
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    • v.2 no.1
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    • pp.41-52
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    • 1996
  • This experiment was carried out to study the effect of sprinkler and fan cooling system on the physiological parameter, milk production and milk composition for Holstein cows in hot, humid climates. Thirty cows were assigned to one of two sections of open-sided loose barn. Water nozzles of sprinkler system were spaced in line at 1.2m intervals. Forced air was provided by 85cm diameter fans at rate of 3.4㎥/sec. The results obtained from these experiments are as follows: 1. There was no significant difference in meteorological data between control and fan + sprinkler cooling system(treatment group). 2. Skin temperature and rectal temperature of the treatment group were significantly lower than those of the control group (32.96 : 39.53$^{\circ}C$ vs 34.02 : 41.21$^{\circ}C$ respectively) (P<0.05). 3. Serum cortisol concentration of the treatment group(0.90$\mu\textrm{g}$/dL) was lower than that of control group(1.44$\mu\textrm{g}$/dL)(P<0.05). 4. Milk production of cows cooled with a sprinkler and fan cooling system was significantly higher than that of no cooling system (P<0.0l). 5. Lactose, protein and solid-not-fat content of milk were not changed by the treatments. Milk fat content of the control(3.23%) was low compared with the treatment group(3.38%). Somatic cell count was reduced by 26.63% in the treatment. The results indicate that a sprinkler and fan cooling systems can provide an effective means to relieve heat stress and enhance productivity of lactating Holstein cows during hot and humid summer season.

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