• Title/Summary/Keyword: Da-Po

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Detection of Chlorotoluene and Nitrotoluene Compounds by Recombinant Microbial Biosensors (재조합 미생물 바이오센서를 이용한 chlorotoluene과 nitrotoluene 화합물의 검출)

  • Lee, Da Young;Cho, Jae Ho;Lim, Woon Ki;Shin, Hae Ja
    • Journal of Life Science
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    • v.24 no.1
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    • pp.54-60
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    • 2014
  • Aromatic hydrocarbons are toxic environmental pollutants that are detrimental to the ecosystem and human health. Among them, chlorotoluene and nitrotoluene are toxic to hydrobios and irritate the skin, eyes, and respiratory organs of humans. We herein report the development of recombinant microbial biosensors for cheap and rapid monitoring of chlorotoluene and nitrotoluene compounds. Plasmids were constructed by inserting the xylR regulatory gene for BTEX (benzene, toluene, ethylbenzene, and xylene) degradation into upstream of Po' (the DmpR activator promoter Po with the deletion of its own upstream activating sequences) or Pu (the cognate promoter of XylR)::lacZ (the ${\beta}$-galactosidase gene) and transformed into Escherichia coli $DH5{\alpha}$. In the presence of inducers, the biosensor cells immobilized in agarose developed a red color in 1-2 h due to the hydrolysis of chlorophenol red ${\beta}$-D-galactopyranoside (CPRG), a substrate of ${\beta}$-galactosidase that was expressed by the inducers. Among BTEX, high responses were specifically observed with o-, m-, p-chlorotoluene ($0.1{\mu}M-100 mM$) and o-, m-, p-nitrotoluene (0.1 mM-100 mM). Po' demonstrated higher responses than those with Pu. The biosensors immobilized in agarose showed good stability after 21 days' storage at $4^{\circ}C$, and responses in untreated wastewater spiked with chlorotoluene and nitrotoluene, suggesting they can be used to detect compounds in wastewater.

Purification of a Protease Produced by Bacillus subtilis PCA 20-3 Isolated from Korean Traditional Meju (전통 메주로부터 분리한 Bacillus subtilis PCA 20-3 유래 Protease 의 정제)

  • Lim, Seong-Il;Yoo, Jin-Young
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1635-1641
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    • 1999
  • Bacillus subtilis PCA20-3 was isolated from meju and was found to produce a protease. The strain produced the maximum amount of enzyme in the medium containing soytone (0.2%), soluble starch (2%), $(NH_4)_2SO_4\;(0.1%),\;CaCl_2(0.1%),\;yeast\;extract\;(0.01%),\;K_2HPO_4\;(0.1%),\;and\;KH_2PO_4\;(0.1%)$. Protease was first concentrated by ammonium sulfate (80% saturation, w/v) precipitation of culture supernatant. Then the enzyme was purified by column chromatography using CM Sephadex C-50. The collected proteins were rechromatographed using Sephadex G-100 gel filtration column. The fraction with protease active from Sephadex G-100 gel chromatography was found to be pure when examined by SDS-polyacrylamide gel electrophoresis and YMC-pak reverse phase chromatography. Specific activity, yield and purity were 76 U/mg. 2.7%, and 7.6 fold, respectively. The molecular weight of the enzyme was estimated to be 31.5 kDa by SDS-PAGE. The number of amino acids calculated from molecular weight was evaluated about 321 residues. N-terminal sequence of the enzyme was $Val^1-Pro^2-Tyr^3-Gly^4-Val^5-Ser^6-Gln^7-Gly^8-Lys^9-Ala^{10}$.

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균일막 형성을 위한 항공기 부품용 타이타늄의 양극산화 최적 공정

  • Lee, Da-Yeong;Han, A-Yeong;Jeong, Na-Gyeom;Choe, Jin-Seop
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2018.06a
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    • pp.66.2-66.2
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    • 2018
  • 금속의 양극산화 공정(anodizing)은 전해질 내 금속에 인위적으로 전위를 가해 금속 표면에 얇은 산화막(oxide layer)을 형성하여 금속의 내식성, 내마모성을 증가시키는 공정이다. 타이타늄은 가볍고 단단하여 산업분야에 유용하게 사용되며 이와 같은 양극산화 공정을 통해 내식성, 내마모성을 크게 높일 수 있다. 본 연구에서는 항공기 부품용 타이타늄의 최적 양극산화 조건을 찾기 위해 전압의 파형, 전해액의 조성에 따라 양극산화 실험을 진행하였다. SEM, AFM, EDS, 분광측색계, 색채색차계 등을 이용하여 각 조건에 해당하는 타이타늄의 산화막($Tio_2$)의 두께, crack 형태, pore 형태, 균일도, 표면 조도, 내전압, 색 수치를 분석하였다. 그 결과 전압 DC 140 V, 주성분이 KOH $Na_3PO_4{\cdot}12H_2O$인 전해액으로 이루어진 양극산화 조건에서 가장 균일하고 색 재현성이 우수한 타이타늄의 산화막($Tio_2$)을 형성하였다.

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Competition between Phase Separation and Crystallization in a PCL/PEG Polymer Blend Captured by Synchronized SAXS, WAXS, and DSC

  • Chuang Wei-Tsung;Jeng U-Ser;Sheu Hwo-Shuenn;Hong Po-Da
    • Macromolecular Research
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    • v.14 no.1
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    • pp.45-51
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    • 2006
  • We conducted simultaneous, small-angle, X-ray scattering/differential scanning calorimetry (SAXS/DSC) and simultaneous, wide-angle, X-ray scattering (WAXS)/DSC measurements for a polymer blend of poly($\varepsilon$-caprolactone)/poly(ethylene glycol)(PCL/PEG). The time-dependent SAXS/DSC and WAXS/DSC results, measured while the system was quenched below the melting temperature of PCL from a melting state, revealed the competitive behavior between liquid-liquid phase separation and crystallization in the polymer blend. The time-dependent structural evolution extracted from the SAXS/WAXS/DSC results can be characterized by the following four stages in the PCL crystallization process: the induction (I), nucleation (II), growth (III), and late (IV) stages. The influence of the liquid-liquid phase separation on the crystallization of PCL was also observed by phase-contrast microscope and polarized microscope with 1/4$\lambda$ compensator.

Development of a Scenario-based GIS Tool for Planning and Design of Eco-Village (시나리오 기반의 생태마을 계획 및 설계 지원 GIS도구 개발)

  • Park, Sung-Woo;Kim, Doo-Soon;Ahn, Kyung-Mo
    • KIEAE Journal
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    • v.13 no.3
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    • pp.41-49
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    • 2013
  • A tool to assist in designing an eco-village was developed using GIS(Geographical Information System) and RS(Remote Sensing) data and images. The difficulties of using GIS by untrained designers are resolved by simplifying the usage and making it a user friendly scenario-based tool, so that the designers with limited knowledge in GIS can use as a design tool. For this task, Da-Mu-Po, a village in Pohang, Gyeongbuk was picked as a site to test the design tool; through planning and designing as an eco friendly village, we tested the flexibility and usability of our newly developed design tool. From this experiment we also introduced test version of stand-alone GIS design tool by constructing program scenario and GUI(Graphic User Interface).

Biochemical analysis of a high-molecular-weight protease from Streptomyces tendae JC412

  • Seong, Chi-Nam;Nam, Doo-Hyun;Kim, Sung-Jun;Cho, Sueung-Sik;Han, Ji-Man;Kim, Joon-Ho;Lee, Hyo-Jung;Yoo, Jin-Cheol
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.333.1-333.1
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    • 2002
  • Streptomyces tendae JC412 secreted two forms of protease(ST -1 and ST -2) when grown in OSY medium (oatmeal 1.5%. soybean meal 2%. dried yeast 1 %) supplemented with glucose(0.5%) and KH2PO4(0.05%). Initial pH of the culture medium was adjusted to 10.0 with NaOH and incubated at $27^{\circ}C$ on a rotary shaking incubator (180rpm). High- molecular-weight protease ST-1 was heat labile. whereas low molecular protease ST-2(22.000 Da) was reported to be heat stable. (omitted)

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Purification and Characterization of Phytase from Bacillus subtilis (Bacillus subtilis가 생산하는 Phytase의 정제 및 특성)

  • Koh Hyun-Jung;Chu In-Ho;Chung Kun-Sub
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.40-46
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    • 2006
  • A bacterial strain producing high level of a phytase was isolated from cattle feces and identified as Bacillus subtilis, and designated as Bacillus sp. CF 5-26. The production of the phytase from Bacillus sp. CF 5-26 reached the highest level after 72 hours at $37^{\circ}C$. The optimum condition of the media for the production of phytase was 10% rice bran extract, 0.1% whey protein powder, $0.01%\;CaCl_{2},\;0.01%\;KH_{2}PO_4$. The phytase was purified 20.3 folds with ethanol precipitation, Sephadex G-100, CM Sepharose CL-6B and Sephacryl S-100-HR column chromatography. The molecular weight of the purified enzyme was estimated to be 66 kDa on SDS-polyacrylamide gel electrophoresis. The purified phytase activity was stable up pH 5.0, 7.0, 11.0 and the remaining activity was 50% when it was treated at $100^{\circ}C$ for 1 hour. The substrate specificity of phytase was most active against sodium phytate and inositol polyphosphate compound. And the phytase hydrolysed tripolyphosphate and pyrophosphate a little. The Km value for the sodium phytate was 0.64 mM and the Vmax value was $4.41\;{\mu}mol/min$.

Purification and Characterization of Bacillus subtilis JS-17 Collagenase. (Bacillus subtilis JS-17이 생산하는 Collagenase의 정제 및 특성)

  • Lim Kyoung-Suk;Son Shung-Hui;Kang Ho Young;Jun Hong-Ki
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.657-663
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    • 2005
  • Collagenases are generally defined as enzymes that are capable of degrading the polypeptide backbone of native collagen under conditions that do not denature the protein. An extracellular collagenase-producing bacterial strain was isolated from kimchi and identified to be Bacillus subtilis JS-17 through morphological, cultural, biochemical characteristics and 16S rDNA sequence analysis. Optimum culture condition of Bacillus subtilis JS-17 for the production of collagenase was $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4\cdot7H_2O,\;0.01\%\; MnSO_4\cdot4H_2O,\;,0.1\%$ citrate and $0.1\%\;CaCl_2$. The production of collagenase was optimal at $30^{\circ}C$ for 72 hr. A collagenase was isolated from the culture filtrate of Bacillus subtilis JS-17. The enzyme was purified using Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography and DEAE-Sephadex A-50 column chromatography The purified collagenase has an specific activity 192.1 units/mg. The molecular weight of the purified enzyme was estimated to be 28 kDa by SDS-PACE. The purified collagenase has $100\%$ activity up to $55^{\circ}C$.

Optimization of Media Composition on the Production of Melanin Bleaching Enzyme from Peniophora sp. JS17 (Peniophora sp. JS17 유래 멜라닌 탈색 효소 생산을 위한 배지 조성의 최적화)

  • Son, Min-Jeong;Kim, Yeon-Hee;Nam, Soo-Wan;Jeon, Sung-Jong
    • Microbiology and Biotechnology Letters
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    • v.47 no.2
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    • pp.250-258
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    • 2019
  • Peniphora sp. JS17, isolated from forest old tree, produced extracellular enzymes that decolorized human hair melanin. The JS17 strain had laccase and manganese peroxidase activity while it did not has lignin peroxidase activity. Batch culture indicated that the melanin decolorization activity of JS17 strain originated from laccase. The culture conditions to maximize the production of melanin bleaching enzymes from Peniophora sp. JS17 mycelia were investigated. Among the tested media for the laccase production, minimal medium (2% glucose, 0.2% malt extract, 0.1% $KH_2PO_4$, 0.4% $MgSO_4{\cdot}7H_2O$) showed the highest activity of laccase. Then, to optimize the culture condition for the laccase activity, the influence of various carbon and nitrogen sources was investigated in minimal medium. Among various carbon and nitrogen sources, 2% xylose and 0.4% tryptone showed the highest production of laccase, respectively. The enzyme was purified using $(NH_4)_2SO_4$ precipitation and Hitrap Q sepharose column, and the purified enzyme showed two isoenzymatic bands with molecular masses of about 70 kDa by SDS-PAGE. The melanin decolorization activity was 77% and 55% within 48 h in the presence of 1-hydroxybenzotriazole (HBT) and syringaldehyde, respectively, whereas only about 9% melanin decolorized in case of no mediator.

Staphylococcus pseudintermedius-induced Aortic and Tricuspid Bacterial Endocarditis in a Dog (개에서 Staphylococcus pseudintermedius에 의해 발생한 심내막염 증례)

  • Kim, Jun-Hwan;Song, Ru-Hui;Lee, Da-Mi;Lee, Hyeon-Suk;Cho, Ho-Seong;Shin, Gee-Wook;Park, Jin-Ho;Park, Chul
    • Journal of Veterinary Clinics
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    • v.29 no.3
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    • pp.255-258
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    • 2012
  • A 14-year-old spayed female, Shihtzu dog, presented with anorexia, depression and respiratory distress. She had a history of mastectomy for mammary gland tumor before 2 weeks. Severe gingivitis, dental plaque, and calculus were confirmed on physical examination. On auscultation, the dog had a diastolic and systolic murmur at the left heart base and right heart apex respectively. The dog had valvular vegetation including tricuspid and aortic valves with regurgitation on echocardiography. Blood culture was performed to confirm bacterial endocarditis and identify pathogens of bacterial endocarditis. Before the result of blood culture was confirmed, antibiotic (cefalexin, 30 mg/kg, PO, q12hr) with furosemide (2 mg/kg, PO, q12hr) and benazepril (0.25 mg/kg, PO, q12hr) were administered empirically and the patient was well controlled. Staphylococcus pseudintermedius was confirmed later. One week later, however, the patient died of acute respiratory distress caused by fulminant pulmonary edema. The owners denied necropsy of the patient.