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Expression of Acetohydroxyacid Synthase from Bacillus anthracis and Its Potent Inhibitors

  • Choi, Kyoung-Jae;Pham, Chien Ngoc;Jung, Hoe-Il;Han, Sung-Hwan;Choi, Jung-Do;Kim, Jin-Heung;Yoon, Moon-Young
    • Bulletin of the Korean Chemical Society
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    • v.28 no.7
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    • pp.1109-1113
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    • 2007
  • Acetohydroxyacid synthase (AHAS, EC 2. 2. 1. 6) is the enzyme that catalyses the first step in the common pathway of the biosynthesis of the branched chain amino acids, valine, leucine and isoleucine. For the first time, the AHAS gene from Bacillus anthracis was cloned into the expression vector pET28a(+), and was expressed in the E. coli strain BL21(DE3). The purified enzyme was checked on 12% SDS-PAGE to be a single band with molecular weight of 65 kDa. The optimum pH and temperature for B. anthracis AHAS was at pH 7.5 and 37 oC, respectively. Kinetic parameters of B. anthracis were as follows: Km for pyruvate, K0.5 for ThDP and Mg2+ was 4.8, 0.28 and 1.16 mM respectively. AHAS from B. anthracis showed strong resistance to three classes of herbicides, Londax (a sulfonylurea), Cadre (an imidazolinone), and TP (a triazolopyrimidine). These results indicated that these herbicides could be used in the search for new anti-bacterial drugs.

Effect of Different Nutrient Solution and Light Quality on Growth and Glucosinolate Contents of Watercress in Hydroponics (배양액의 종류 및 광질이 물냉이의 생육 및 Glucosinolate 함량에 미치는 영향)

  • Choi, Jae Yun;Kim, Sung Jin;Bok, Kwon Jeong;Lee, Kwang Ya;Park, Jong Seok
    • Journal of Bio-Environment Control
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    • v.27 no.4
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    • pp.371-380
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    • 2018
  • Aim of this study was to investigate the effects of different nutrient solutions and various light qualities generated by LED on the growth and glucosinolates contents of watercress (Nasturtium officinale) grown under hydroponics for 3 weeks. The seeds of watercress were sown on crushed rockwool media and raised them for two weeks. They were transplanted in a semi-DFT (deep flow technique) hydroponics system. A controlled-environment room was maintained at $20{\pm}1^{\circ}C$ and $16{\pm}1^{\circ}C$ temperatures and $65{\pm}10%$ and $75{\pm}10%$ relative humidity (day and night, respectively), with a provided photosynthetic photon flux density (PPFD) of $180{\pm}10{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ and a photoperiod of 16/8h. To find out the best kinds of nutrient solutions for growing watercress, Otsuka House 1A (OTS), Horticultural Experiment Station in Korea (HES), and Netherland's Proefstaion voor Bloemisterij en Gasgroente (PBG) were adapted with initial EC of $1.0-1.3dS{\cdot}m^{-1}$ and pH of 6.2, irradiating PPFD with fluorescent lamps (Ex-1). Either monochromatic (W10 and R10) or mixed LEDs (R5B1, R3B1, R2B1G1, and W2B1G1) were irradiated with a differing ratio of each LED's PPFD to understanding light quality on the growth and glucosinolates contents of watercress (Ex-2). Although significant difference in the shoot growth of watercress was not found among three nutrient solutions treatments, but the root fresh weight increased by 13.7% and 55.1% in PBG and OTS compared to HES, respectively. OTS increased the gluconasturtiin content by 96% and 65% compared to PBG and HES. Compared with the white light (W10), the red light (R10) showed a 101.3% increase in the shoot length of watercress. Increasing blue light portion positively affected plant growth. The content of total glucosinolates in watercress was increased by 144.5% and 70% per unit dry weight in R3B1 treatment compared with R2B1G1 and W10 treatments, respectively. The growth and total glucosinolates contents of the watercress were highest under R3B1 among six light qualities.

2'-Spirocyclopropyl-carbocyclic Nucleoside as a Novel Scaffold for Potent Anti-HCV Agents

  • Li, Hua;Yoo, Jin-Cheol;Hong, Joon-Hee
    • Bulletin of the Korean Chemical Society
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    • v.32 no.4
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    • pp.1146-1152
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    • 2011
  • The discovery of 2'-spirocyclopropyl-ribocytidine (J. Med. Chem. 2010, 53, 8150-8160) as a potent inhibitor of RNA synthesis by NS5B ($IC_{50}=7.3{\mu}M$), the RNA polymerase encoded by hepatitis C Virus (HCV), has led to the synthesis and biological evaluation of several carbocyclic versions of 2'-spiropropyl-nucleosides. The cyclopentenol intermediate 7 was successfully constructed via ring-closing metathesis (RCM) from divinyl 6. Spirocyclopropanation of enone 8 was effected by using (2-chloroethyl)-dimethylsulfonium iodide and potassium tert-butoxide to form the desired intermediate 9. The synthesized nucleoside analogues 21-24 were assayed for their ability to inhibit HCV RNA replication in a subgenomic replicon Huh7 cell line. Among them, the cytosine nucleoside analogue 22 exhibited significant anti-HCV activity ($EC_{50}= 8.2{\mu}M$).

Light Quality and Photoperiod Affect Growth of Sowthistle (Ixeris dentata Nakai) in a Closed-type Plant Production System (밀폐형 식물생산시스템에서 광질과 광주기에 따른 씀바귀의 생육)

  • Kim, Hye Min;Kang, Jeong Hwa;Jeong, Byoung Ryong;Hwang, Seung Jae
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.67-76
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    • 2016
  • This study was conducted to examine the optimal environmental condition for promoting the growth of sowthistle as affected by light quality and photoperiod in a closed-type plant production system. Seeds were sown in 240-cell plug trays and then germinated for 3 days at a 24-hour photoperiod in a closed-type plant production system with LED lights (R:B:W = 8:1:1). Seedlings were transplanted and grown under 3 types of LED (R:B:W = 8:1:1, R:W = 3:7, or R:B = 8:2) and 4 photoperiods (24/0, 16/8, 8/16, or 4/20 hours) with $230{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ light intensity at a density of $20cm{\times}20 cm$. The experimental design was a randomized complete block design. Plants were cultured for 40 days un der the condition of $21{\pm}2^{\circ}C$ and $70{\pm}10%$ relative humidity after transplanting. Plants were fed with a recycling nutrient solution (pH 7.0 and EC $2.0dS{\cdot}m^{-1}$) contained in a deep floating tank. Fresh weight and dry weight of shoot or root, leaf length, and leaf area were the greatest in the photoperiod of 24/0 (light/dark) with RW LED. The highest number of leaves occurred in the photoperiod of 16/8 (light/dark) with RB LED, while the incidence of tip burn was higher in the photoperiod of 24/0 (light/dark) compared to the other treatments. Chlorophyll value was the highest in the 16/8 (light/dark) photoperiod and there was no significant difference by light quality. Chlorophyll fluorescence was the lowest in the photoperiod of 24/0 (light/dark) compared with other treatments. Therefore, in terms of economic feasibility and productivity for Ixeris dentata Nakai cultivation in a closed-type plant production system, the results obtained suggest that plants grew the best when kept in a photoperiod of 16/8 (light/dark) and light quality of combined LED RW (3:7).

Effects of Water-logging on the Chemical Properties, Microflora and Biomass in Continuous Cropping of Cucumber Soils (오이연작토양(連作土壤)의 화학성(化學性), 미생물상(微生物相) 및 Biomass 에 미치는 담수처리(湛水處理)의 영향(影響))

  • Kim, Hyun-Woo;Kim, Young-Woong;Kim, Kwang-Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.22 no.2
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    • pp.146-155
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    • 1989
  • A series of the experiments were conducted to evaluate the effects of the water-logging in continuous cropping soils of cucumber. Some of the results obtained from these experiments are summarized as follow: The values of pH in water-logged treatments were from 5.50 to 5.90. Those of EC were less than 0.3 mmhos/cm, were lower than those of control plot. The ratios of Ca/Mg, Ca/K, Mg/K respectively 2.5, 4.8, 2.0, were similar to the standard values. These changes were suitable for cucumber planting. And the number of bacteria among the microflora were determined increased from $10^6$ to $10^7\;cells/g$ dry soil, while that of the Fungi, Fusarium spp, Pythium spp, and Phythophthora spp decreased. In proportion to these changes of microflora, the ratios of B/F in water-logging plot were higher than in the control plot. The contents of Biomass-C were showed 10.47mg/100g soil to 36.12 mg/100g soil, and the contents of that in water-logging were more than in control plots.

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Variation of AC Impedance of the $TiS_2$ Composite/SPE/Li Cell with Cycling ($TiS_2$ Composite/SPE/Li Cell의 충방전에 따른 AC 임피던스의 변화)

  • Kim, J.U.;Gu, H.B.;Moon, S.I.;Yun, M.S.
    • Proceedings of the KIEE Conference
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    • 1995.07c
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    • pp.1034-1038
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    • 1995
  • The purpose of this study is to research and develop $TiS_2$ composite cathode for lithium polymer battery(LPB). $TiS_2$ electrode represent a class of insertion positive electrode used in Li secondary batteries. In this study, we investigated preparation of $TiS_2$ composite cathode and AC impedance response of $TiS_2$ composite/SPE/Li cells as a function of state of charge(SOC) and cycling. The resistance of B type cell using $TiS_2PEO_8LiClO_4PC_5EC_5$ composite cathode was lower than that of A type cell using $TiS_2PEO$ composite cathode. The cell resistance of B type cell is high for the first few percent discharge, decreases for midium discharge and then increases again toward the end of discharge. We believe the magnitude of the cell resistance is dominated by passivation layer impedance and small cathode resistance. AC impedance results indicate that the cell internal resistance increase with cycling, and this is attributed to change of passivation layer impedance with cycling. The passivation layer resistance($R_f$) of B type cell decreases for the 2nd cycling and then increases again with cycling. Redox coulombic efficiency of B type cell was about 141% at 1st cycle and 100% at 12th cycle. Also, $TiS_2$ specific capacity was 115 mAh/g at 12 cycle.

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The Preparation and Characterization of Bismuth Layered Ferroelectric Thin Films by Sol-Gel Process (II. Dielectric Properties of Ferroelectric $Sr_{0.7}/B_{2.3}(Ta_{1-x}Nb_x)_2O_9$ Thin Films Prepared by MOD Process) (솔 - 젤법을 이용한 Bismuth Layered Structure를 가진 강유진성 박막의 제조 및 특성평가에 관한 연구 (II. MOD법으로 제조한 강유전성 $Sr_{0.7}/B_{2.3}(Ta_{1-x}Nb_x)_2O_9$ 박막의 유전특성))

  • 최무용;송석표;정병직;김병호
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.12 no.1
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    • pp.62-68
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    • 1999
  • Ferroelectric $Sr_{0.7}/B_{2.3}(Ta_{1-x}Nb_x)_2O_9$(x=0, 0.1, 0.2, 0.3) thin films were deposited on $Pt/SiO_2/Si$ substrate by MOD(Metalorganic Decomposition) process. Metal carboxylate and metal alkoxide were used as precursors, and 2-methoxyethanol, xylene as solvents. After spin coating, thin films were pre-annealed at $400^{\circ}C$, followed by RTA(Rapid Thermal Annealing) and final annealing at $800^{\circ}C$ in oxygen atmosphere. These procedures were repeated three times to obtain thin films with the thickness of $2000{\AA}$. To enhance the nucleation and growth of layered-perovskite phase, thin films were rapid-thermally annealed above $720^{\circ}C$ in oxygen atmosphere. As RTA temperature increased, fluorite phase was transformed to layered-perovskite phase. And the change of Nb contents affected dielectric / electrical properties and microstructure. The ferroelectric characteristics of $Sr_{0.7}/B_{2.3}(Ta_{1-x}Nb_x)_2O_9$ thin film were Pr=8.67 $\mu{C}/cm^2$, Ec=62.4kV/cm and $I_{L}=1.4\times10^{-7}A/cm^2$ at the applied voltage of 5V, respectively.

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Doherty Amplifier Using Load Modulation and Phase Compensation DGS Micro-Strip Line (부하 변조 및 위상 보상 DGS 마이크로스트립 선로를 이용한 도허티 증폭기)

  • Choi Heung-Jae;Lim Jong-Sik;Jeong Yong-Chae
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.16 no.8 s.99
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    • pp.815-824
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    • 2005
  • In this paper, we proposed a new DGS(Defected Ground Structure) Doherty amplifier for IMT-2000 band. Originally, active load-pull analysis of a Doherty amplifier assumes ideal harmonic termination condition. However, there have been no papers considering this ideal harmonic termination condition. We obtained excellent improvements of efficiency, gain, maximum output power as well as superior size reduction of a Doherly amplifier by satisfying the overlooked assumption of ideal harmonic termination through the adaptation of DGS at the output transmission line of carrier and peaking amplifier that is essential for Doherty operation. The amount of both the 2nd and the 3rd harmonic rejection of the proposed DGS Doherty amplifier over the conventional one are 44.92 dB and over 23.77 dB, respectively. The acquired improvement in Pl dB, gain, drain efficiency, and ACPR to WCDMA 1FA signal were 0.42 dB, 0.33 dB, $6.4\%$ and 5.4 dBc, respectively. Moreover, electrical length of $90{\circ}$ is reduced at each of the DGS carrier amplifier path and DGS peaking amplifier path, therefore the whole amplifier circuit size is considerably reduced.

A Study on the Safety of Mycotoxins in Grains and Commonly Consumed Foods (곡류 등 다소비 식품 중 곰팡이독소 안전성 조사 연구)

  • Kim, Jae-Kwan;Kim, Young-Sug;Lee, Chang-Hee;Seo, Mi Young;Jang, Mi Kyung;Ku, Eun-Jung;Park, Kwang-Hee;Yoon, Mi-Hye
    • Journal of Food Hygiene and Safety
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    • v.32 no.6
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    • pp.470-476
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    • 2017
  • The purpose of this study was to investigate and evaluate the safety of the grains, nut products, beans and oilseeds being sold in Gyeonggi province by analyzing mycotoxins. A multi-mycotoxins analysis method based on LC-MS/MS was validated and applied for the determination of eight mycotoxins, including aflatoxins ($B_1$, $B_2$, $G_1$ and $G_2$), fumonisins ($B_1$, $B_2$), zearalenone and ochratoxcin A in 134 samples. The limit of detection (LOD) and limit of quantitation (LOQ) for the eight mycotoxins ranged from 0.14 to $8.25{\mu}g/kg$ and from 1.08 to $7.21{\mu}g/kg$, respectively. Recovery rates of mycotoxins were determined in the range of 61.1 to 97.5% with RSD of 1.0~14.5% (n=3). Fumonisin $B_1$, $B_2$, zearalenone, and ochratoxin A were detected in 22 samples, indicating that 27% of grains, 12.5% of beans and 11.8% of oilseeds were contaminated. Fumonisin and zearalenone were detected simultaneously in 2 adlays and 3 sorghums. Fumonisin $B_1$ and $B_2$ were detected simultaneously in most samples whereas fumonisin $B_1$ was detected in 1 adlay, 1 millet and 1 sesame sample. The average detected amount of fumonisin was $49.3{\mu}g/kg$ and $10.1{\mu}g/kg$ for grains and oilseeds, respectively. The average detected amount of zearalenone was $1.9{\mu}g/kg$ and $1.5{\mu}g/kg$ for grains and beans, respectively. In addition, the average amount of ochratoxin A was $0.08{\mu}g/kg$ for grains. The calculated exposure amounts of fumonisin, zeralenone and ochratoxin A for grains, beans and oilseeds were below the PMTDI/PTWI.

Cloning and Characterization of a Methionine Aminopeptidase (MAP) Gene from Tetragenococcus halophilus CY54 Isolated from Myeolchi-Jeotgal

  • Tae Jin Kim;Min Jae Kim;Yun Ji Kang;Ji Yeon Yoo;Jeong Hwan Kim
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.26-31
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    • 2023
  • A map gene encoding methionyl-specific aminopeptidase (MAP; EC 3.4.11.18) was cloned from Tetragenococcus halophilus CY54. Translated amino acid sequence of CY54 MAP showed high similarities with those from Enterococcus faecalis (83.8%) and Streptococcus salivarius (62.2%) but low similarities with MAPs from Lactobacillus and Lactococcus genera. The map gene was overexpressed in E. coli BL21(DE3) using pET26b(+),pET26b(+), and the recombinant MAP was purified by using an Ni-NTA column. The size of recombinant MAP was 29 kDa as determined by SDS-PAGE. The optimum pH and temperature of CY54 MAP were pH 5.0 and 60℃, respectively. The activity of CY54 MAP was most significantly increased by Co2+ ion (159%), and showed the highest activity at 12% NaCl. Km and Vmax were 0.64 ± 0.006 mM and 10.12 ± 0.014 U/mg protein, respectively when met-pNA was used as the substrate. This is the first report on a MAP from Tetragenococcus species.