• Title/Summary/Keyword: Fusion temperature

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Purification of Human HtrA1 Expressed in E. coli and Characterization of Its Serine Protease Activity (E. coli에서 발현된 human HtrA1 단백질의 정제와 HtrA1의 serine protease 활성 조건에 관한 연구)

  • Kim, Kyung-Hee;Kim, Sang-Soo;Kim, Goo-Young;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1133-1140
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    • 2006
  • Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.

Analysis of Surface Urban Heat Island and Land Surface Temperature Using Deep Learning Based Local Climate Zone Classification: A Case Study of Suwon and Daegu, Korea (딥러닝 기반 Local Climate Zone 분류체계를 이용한 지표면온도와 도시열섬 분석: 수원시와 대구광역시를 대상으로)

  • Lee, Yeonsu;Lee, Siwoo;Im, Jungho;Yoo, Cheolhee
    • Korean Journal of Remote Sensing
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    • v.37 no.5_3
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    • pp.1447-1460
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    • 2021
  • Urbanization increases the amount of impervious surface and artificial heat emission, resulting in urban heat island (UHI) effect. Local climate zones (LCZ) are a classification scheme for urban areas considering urban land cover characteristics and the geometry and structure of buildings, which can be used for analyzing urban heat island effect in detail. This study aimed to examine the UHI effect by urban structure in Suwon and Daegu using the LCZ scheme. First, the LCZ maps were generated using Landsat 8 images and convolutional neural network (CNN) deep learning over the two cities. Then, Surface UHI (SUHI), which indicates the land surface temperature (LST) difference between urban and rural areas, was analyzed by LCZ class. The results showed that the overall accuracies of the CNN models for LCZ classification were relatively high 87.9% and 81.7% for Suwon and Daegu, respectively. In general, Daegu had higher LST for all LCZ classes than Suwon. For both cities, LST tended to increase with increasing building density with relatively low building height. For both cities, the intensity of SUHI was very high in summer regardless of LCZ classes and was also relatively high except for a few classes in spring and fall. In winter the SUHI intensity was low, resulting in negative values for many LCZ classes. This implies that UHI is very strong in summer, and some urban areas often are colder than rural areas in winter. The research findings demonstrated the applicability of the LCZ data for SUHI analysis and can provide a basis for establishing timely strategies to respond urban on-going climate change over urban areas.

Biochemical Characterization of Heterologously Expressed Chitinase 1 (Chi1) from an Inky Cap, Coprinellus congregatus (이형 재조합한 먹물버섯 Coprinellus congregatus Chitinase 1 (Chi1)의 발현과 생화학적 특성 분석)

  • Yoo, Yeeun;Choi, Hyoung T.
    • Korean Journal of Microbiology
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    • v.49 no.4
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    • pp.309-312
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    • 2013
  • Fungal cell walls consist of various glucans and chitin. Fungi produce chitinases for their growth and development. The inky cap, Coprinellus congregatus, produces at least two different chitinases during its life cycle. Chitinase 1 (chi1) is expresses throughout its life cycle while chitinase 2 (chi2) is expressed at the mushroom autolysing phase. The cloned cDNA of chi1 is successfully expressed as a fusion protein with c-myc in Pichia pastoris, and purified by the affinity chromatography. The optimum pH and temperature of Chi1 was pH 8.0 and $35^{\circ}C$, respectively when 4-nitrophenyl N,N',N"-triacetyl-${\beta}$-D-chitotrioside was used as the substrate. The $K_m$ value and $V_{max}$ for the substrate was 0.780 mM and 0.10 OD $min^{-1}unit^{-1}$, respectively. The addition of purified Chi1 resulted in total growth inhibition against several plant pathogenic fungi such as Alternaria alternata, Fusarium graminearum and Trichoderma harzianum at the concentration of 60 ${\mu}g/ml$.

A Study on the Warpage of Glass Fiber Reinforced Plastics for Part Design and Operation Condition: Part 2. Crystalline Plastics (유리섬유로 보강된 수지에서 제품설계 및 성형조건에 따른 휨의 연구: Part 2. 결정성 수지)

  • Lee, Min;Kim, Hyeok;Lyu, Min-Young
    • Polymer(Korea)
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    • v.36 no.6
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    • pp.677-684
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    • 2012
  • Injection molding process is a popular polymer processing involving plasticizing and enforcing the material flow into the mold. A polymer material shrinks according to temperature variations during the shaping process, and subsequently molding shrinkage developed. Developed deflections or warpages after molding process in part are caused by residual stress relaxation contained in the part. Adding inorganic materials or fibers such as glass and carbon to control shrinkage and enhance warpage resistance are common. In this study, warpages according to part design have been investigated through experiment. Warpages for molding conditions and mold designs such as gate locations were measured. Warpages along flow direction and perpendicular to the flow direction were also measured. Warpages near gate and far from gate were compared. Glass fiber reinforced crystalline polymers, PP and PA66 have been used in this experiment. Glass fiber reinforced crystalline polymers showed large warpage compared with glass reinforced amorphous polymers. Warpages in crystalline polymers were less influenced by molding conditions compared with amorphous polymers, however warpages of crystalline polymers significantly depend on part design.

Microscopic Analysis of Effect of Shot Peening on Corrosion Fatigue Behavior of Aluminum Alloy (알루미늄합금 재료의 부식피로거동에 미치는 쇼트피닝 효과에 대한 미시적 분석)

  • Kim, Jong-Cheon;Cheong, Seong-Kyun
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.36 no.11
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    • pp.1381-1389
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    • 2012
  • The object of this study considers corrosion fatigue improvement of 7075-T6 aluminum by using shot peening treatment on 3.5% NaCl solution at room temperature. Aluminum alloy is generally used in aerospace structural components because of the light weight and high strength characteristics. Many studies have shown that an aluminum alloy can be approximately 50% lighter than other materials. Mostly, corrosion leads to earlier fatigue crack propagation under tensile conditions and severely reduces the life of structures. Therefore, the technique to improve material resistance to corrosion fatigue is required. Shot peening technology is widely used to improve fatigue life and other mechanical properties by induced compressive residual stress. Even the roughness of treated surface causes pitting corrosion, the compressive residual stress, which is induced under the surface layer of material by shot peening, suppresses the corrosion and increases the corrosion resistance. The experimental results for shot peened specimens were compared with previous work for non treated aluminum alloy. The results show that the shot peening treatment affects the corrosion fatigue improvement of aluminum alloys and the induced compressive residual stress by shot peening treatment improves the resistance to corrosion fatigue.

Study on ICT convergence in Lentinula edodes (Shiitake) cultivation system using Automated container (컨테이너형 수출용 버섯식물공장시스템설계 및 표고버섯 생산 연구)

  • Jo, Woo-Sik;Lee, Sung-Hak;Park, Woo-Ram;Shin, Seung-Ho;Park, Chang-Min;Oh, Ji-Hyun;Park, Who-Won
    • Journal of Mushroom
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    • v.15 no.4
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    • pp.264-268
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    • 2017
  • In the 21st century, information and communication technology (ICT) worldwide presents a new vision for agriculture. Time and place, as well as the high-tech industry, to overcome barriers to the fusion of the so-called "smart agriculture," are changing the agricultural landscape. Core container production in precision agriculture for mushroom cultivation, optimal temperature, humidity, irradiation, self-regulation of factors such as carbon dioxide, and environment for mushroom cultivation were adopted. Lentinula edodes (shiitake) is an edible mushroom native to East Asia, cultivated and consumed in many Asian countries. It is considered to be medicinal in certain practices of traditional medicine. We used different controlled light sources (Blue-Red-White-combined LED, blue LED, red LED, and fluorescent light) with different LED radiation intensities (1.5, 10.5, and $20.5{\mu}mol/m^2s$ for LEDs) to compare growth and development. Mushrooms were treated with light in a 12-hour-on/12-hour-off cycle, and maintained in a controlled room at $19{\sim}21^{\circ}C$, with 80~90% humidity, and an atmospheric $CO_2$ concentration of 1,000 ppm for 30 days. Growth and development differed with the LED source color and LED radiation intensity. Growth and development were the highest at $10.5{\mu}mol/m^2s$ of blue LED light. After harvesting the fruit bodies, we measured their weight and length, thickness of pileus and stipe, chromaticity, and hardness. The $10.5{\mu}mol/m^2s$ blue-LED-irradiated group showed the best harvest results with an average individual weight of 39.82 g and length of 64.03 mm, pileus thickness of 30.85 mm and pileus length of 43.22 mm, and stipe thickness of 16.96 mm with fine chromaticity and hardness. These results showed that blue LED light at $10.5{\mu}mol/m^2s$ s exerted the best effect on the growth and development of L. edodes (shiitake) mushroom in the ICT-system container-type environment.

Combustion characteristics of two imported Indonesia coals as a pulverized fuel of thermal power plants (인도네시아산 발전용 수입 석탄 2종의 연소특성 비교 평가)

  • Lee, Hyun-Dong;Kim, Jae-Kwan
    • Journal of Energy Engineering
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    • v.19 no.2
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    • pp.136-142
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    • 2010
  • Combustion reactivity and thermal behavior of two imported coals used as a pulverized fuel of commercially thermal power plant were investigated by thermogravimetric analysis (TGA) and large scale test furnace of 200 kg/hr. TGA results showed that combustion efficiency of high moisture coal has lower than reference coal due to the slow combustion completion rate although it has the low ignition temperature, and activation energies of high moisture coal with 79 kJ/mol for overall combustion was higher than reference coal of 53 kJ/mol. Test furnace results ascertained that flame of black band of high moisture coal during the combustion in boiler broke out compared to reference coal and then it becomes to unburned carbon due to the less reactivity and combustion rate. But, Blending combustion of high moisture coal with design coal of high sulfur are available because sulfur content of high moisture coal was too low to generate the low SOx content in flue gas from boiler during the combustion. The ash analysis results show that it was not expected to be associated with slagging and fouling in pulverized coal fired systems due to the low alkali metal content of $Na_2O$ and $K_2O$ compared to bituminous coal.

Effects of oxygen partial pressure on the properties of indium tin oxide film on PET substrates by RF magnetron sputtering (RF 마그네트론 스퍼터링법에 의해 PET 기판 위에 증착된 ITO 박막의 특성에 대한 산소 분압의 영향)

  • Kim, Seon Tae;Kim, Tae Gyu;Cho, Hyun;Kim, Jin Kon
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.24 no.6
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    • pp.252-255
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    • 2014
  • Indium tin oxide (ITO) films with various oxygen partial pressure from 0 to $6{\times}10^{-5}$ Pa were prepared onto polyethylene terephthalate (PET) using RF magnetron sputtering at room temperature. The structural, electrical and optical properties of the grown ITO films were investigated as a function of the oxygen partial pressure. The amorphous nature of the ITO films was dominant at the partial pressure below $1{\times}10^{-5}$ Pa and the degree of crystallinity increased as the oxygen concentration increased further. This structural change comes with the increased carrier concentration and reduction of the electrical resistivity down to $9.8{\times}10^{-4}{\Omega}{\cdot}cm$. The average transmittance (at 400~800 nm) of the ITO deposited on the PET substrates increased as the oxygen partial pressure increased and transmittance above 80 % was achieved with the partial pressure of $4{\times}10^{-5}$ Pa. The results show that the choice of optimal oxygen partial pressure can present improved film crystallinity, the increased carrier concentration, and the enhancement in the electrical conductivity.

Biosynthesis of Isoprenoids: Characterization of a Functionally Active Recombinant 2-C-methyl-D-erythritol 4-phosphate Cytidyltransferase (IspD) from Mycobacterium tuberculosis H37Rv

  • Shi, Wenjun;Feng, Jianfang;Zhang, Min;Lai, Xuhui;Xu, Shengfeng;Zhang, Xuelian;Wang, Honghai
    • BMB Reports
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    • v.40 no.6
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    • pp.911-920
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    • 2007
  • Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the leading infectious diseases to humans. It is urgent to discover novel drug targets for the development of antitubercular agents. The 2-C-methyl-Derythritol-4-phosphate (MEP) pathway for isoprenoid biosynthesis has been considered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammals. MEP cytidyltransferase (IspD), the third-step enzyme of the pathway, catalyzes MEP and CTP to form 4-diphosphocytidyl-2-C-methylerythritol (CDP-ME) and PPi. In the work, ispD gene from M. tuberculosis H37Rv (MtIspD) was cloned and expressed. With N-terminal fusion of a histidine-tagged sequence, MtIspD could be purified to homogeneity by one-step nickel affinity chromatography. MtIspD exists as a homodimer with an apparent molecular mass of 52 kDa. Enzyme property analysis revealed that MtIspD has high specificity for pyrimidine bases and narrow divalent cation requirements, with maximal activity found in the presence of CTP and $Mg^{2+}$. The turnover number of MtIspD is $3.4 s^{-1}$. The Km for MEP and CTP are 43 and $92{\mu}M$, respectively. Furthermore, MtIspD shows thermal instable above $50^{\circ}C$. Circular dichroism spectra revealed that the alteration of tertiary conformation is closely related with sharp loss of enzyme activity at higher temperature. This study is expected to help better understand the features of IspD and provide useful information for the development of novel antibiotics to treat M. tuberculosis.

Investigation on Combustion Characteristics of Sewage Sludge using Pilot-scale Bubbling Fluidized Bed Reactor (파일럿 규모 기포 유동층 반응기를 이용한 하수 슬러지 연소 특성 분석)

  • Kim, Donghee;Huh, Kang Y.;Ahn, Hyungjun;Lee, Youngjae
    • Clean Technology
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    • v.23 no.3
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    • pp.331-342
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    • 2017
  • To estimate the combustion characteristics of sewage sludge and wood pellet, thermogravimetric analysis (TGA) was conducted. As TGA results, combustion characteristics of sewage sludge was worse than wood pellet. In ash fusion temperature (AFT) analysis, slagging tendency of sewage sludge is very high compared to wood pellet. And also, the bubbling fluidized bed reactor with a inner diameter 400 mm and a height of 4300 mm was used for experimental study of combustion characteristics fueled by sewage sludge and wood pellet. The facility consists of a fluidized bed reactor, preheater, screw feeder, cyclone, ash capture equipment and gas analyzer. The thermal input of sewage sludge cases were $54.5{\sim}96.5kW_{th}$, in case of wood pellet experiment, it was $96.1kW_{th}$. As experiment results, the $NO_x$ emission of sewage sludge was averagely about 10 times the $NO_x$ emission of wood pellet. And also CO emission of sewage sludge is about 3.5 times of wood pellet. Lastly as a result of analysis of captured ash in cyclone, the combustion efficiency of all cases were over 99%, but the potential for slagging/fouling was high at all cases by component analysis of ash.