• Title/Summary/Keyword: TRC

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Microbial production of carotenoids for fortification of foods

  • Kim, Seon-Won;Keasling, J.D.
    • Proceedings of the Korean Society of Life Science Conference
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    • 2001.11a
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    • pp.3-8
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    • 2001
  • Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all carotenoids, IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of 1-deoxy-D-xylulose-5-phosphate (DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoisomerase and encoded by dxr. To determine if one or more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains (DH5(, XL1-Blue, and JM101) that had been engineered to produce lycopene. Lycopene production was improved significantly in strains transformed with the dxs expression vectors. When the dxs gene was expressed from the arabinose-inducible araBAD promoter (PBAD) on a medium-copy plasmid, lycopene production was 2-fold higher than when dxs was expressed from the IPTG-inducible trc and lac promoters (Ptrc and Plac, respectively) on medium-copy and high-copy plasmids, Given the low final densities of cells expressing dxs from IPTG-inducible promoters, the low lycopene production was probably due to the metabolic burden of plasmid maintenance and an excessive drain of central metabolic intermediates. At arabinose concentrations between 0 and 1.33 mM, cells expressing both dxs and dxr from PBAD on a medium-copy plasmid produced 1.4 - 2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene production in cells expressing both dxs and dxr was lower than in cells expressing dxs only. A comparison of the three E. coli strains transformed with the arabinose-inducible dxs on a medium-copy plamid revealed that lycopene production was highest in XL1-Blue.

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The Two-Component Protease NS2B-NS3 of Dengue Virus Type 2: Cloning, Expression in Escherichia coli and Purification of the NS2B, NS3(pro) and NS2B-NS3 Proteins

  • Champreda, Veerawat;Khumthong, Rabuesak;Subsin, Benchamas;Angsuthanasombat, Chanan;Panyim, Sakol;Katzenmeier, Gerd
    • BMB Reports
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    • v.33 no.4
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    • pp.294-299
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    • 2000
  • Proteolytic processing of the dengue virus serotype 2 polyprotein precursor is catalyzed by a host signal peptidase and a virus encoded two-component protease consisting of the nonstructural proteins, NS2B and NS3. We expressed in Escherichia coli the NS2B, NS3(pro) and NS2B-NS3 proteins from the dengue virus type 2 strain 16681 as N-terminal fusions with a hexahistidine affinity tag under the control of the inducible trc promoter. All fusion proteins were purified to >90% purity by detergent extraction of inclusion bodies and a single step metal chelate chromatography. Proteins were refolded on-column and recovered with yields of 0.5, 6.0 and 1.0 mg/l of E. coli culture that was grown to $OD_{600}=1.0$ for NS2B, NS3(pro) and NS2B-NS3, respectively. Purified proteins gave strong signals in Western blots using $Ni^{2+}-nitrilotriacetic$ acid as a probe for the presence of the polyHis tag. During the purification process, $(His)_{6}NS2B-NS3$ was apparently not autoproteolytically cleaved at the NS2B/NS3 site.

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Isolation, Physicochemical Properties and Toxicities of Territrem A' and B'

  • Ling, Kuo Huang;Peng, Fu Chuo;Chen, Bai Jiun;Wang, Yu;Lee, Gene Hsiang
    • Korean Journal of Pharmacognosy
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    • v.17 no.2
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    • pp.153-160
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    • 1986
  • We have isolated two new metabolites of territrem, designated as territrem $A'\;(TRA';\;C_{28}H_{30}O_{10})$ and $B'\;(TRB';\; C_{29}H_{34}O_{10})$ from chloroform extract of rice culture of Aspergillus terreus 23-1, using the same isolation procedure as that for territrem A, B and C(TRA, TRB, TRC). The present isolation procedure gave about 5 mg of TRA' and 10 mg of TRB' from 4 kg of rice culture per batch. Analysis of the high resolution mass spectrum showed that the molecular composition of TRA' and TRB' are $C_{28}H_{30}O_{10}$ and $C_{29}H_{34}O_{10}$ respectively, Some results of physicochemical properties were presented in this paper. Single crystal X-ray diffractometry of TRB' showed that the three dimensional structure of TRB' has not changed significantly from that of $TRB\;(C_{29}H_{34}O_9)$, except for the insertion of one oxygen atom into TRB to make additional pyran in the E-ring. It is also suggested that the aromatic moiety of TRA' is similar to that of $TRA\;(C_{28}H_{30}O_9)$ and the rest non-aromatic portions resemble to those of TRB'. The tremorgenic activity, lethality and inhibitory effect on acetylcholine esterase of TRA' and TRB' are greatly reduced comparing to that of TRA and TRB.

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Investigation of a Left-Turn Phase Time Estimation Method for TRC Operation (실시간 신호시스템의 좌회전 신호시간 추정방법에 관한 연구 (검지기 장애발생시를 중심으로))

  • An, Hye-Jin;Nam, Baek;Lee, Sang-Su
    • Journal of Korean Society of Transportation
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    • v.25 no.5
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    • pp.33-42
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    • 2007
  • The current left-turn split model adopted in COSMOS has an inherent limitation when a loop detector in the left-turn lanes was disconnected for a period of time. In this instance, the current model always allocated minimum green time to the left-turn phase, thus optimal split and efficient signal operation for the intersection was not guaranteed. In this paper, four mathmatical models using detector information of the intersection and four empirical models using historical profiles were developed and investigated for different traffic conditions to improve the operational efficiency of the intersection. From the model evaluation test, the empirical model using a four-week historical profile produced the least error among the eight models investigated. NETSIM simulation test results also showed that the proposed model could give significantly reduced delay time as compared to the current model. From these results, the operational efficency of the signalized intersections under the real-time control can be greatly improved by using the model proposed in case of the left-turn detector failure.

Monitoring the failure mechanisms of a reinforced concrete beam strengthened by textile reinforced cement using acoustic emission and digital image correlation

  • Aggelis, Dimitrios G.;Verbruggen, Svetlana;Tsangouri, Eleni;Tysmans, Tine;Van Hemelrijck, Danny
    • Smart Structures and Systems
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    • v.17 no.1
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    • pp.91-105
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    • 2016
  • One of the most commonly used techniques to strengthen steel reinforced concrete structures is the application of externally bonded patches in the form of carbon fiber reinforced polymers (CFRP) or recently, textile reinforced cements (TRC). These external patches undertake the tensile stress of bending constraining concrete cracking. Development of full-field inspection methodologies for fracture monitoring are important since the reinforcing layers are not transparent, hindering visual observation of the material condition underneath. In the present study acoustic emission (AE) and digital image correlation (DIC) are applied during four-point bending tests of large beams to follow the damage accumulation. AE helps to determine the onset of fracture as well as the different damage mechanisms through the registered shifts in AE rate, location of active sources and change in waveform parameters. The effect of wave propagation distance, which in large components and in-situ can well mask the original information as emitted by the fracture incidents is also discussed. Simultaneously, crucial information is supplied by DIC concerning the moments of stress release of the patches due to debonding, benchmarking the trends monitored by AE. From the point of view of mechanics, conclusions on the reinforcing contribution of the different repair methodologies are also drawn.

Changes in Microstructure and Mechanical Properties due to Heat Treatment of Mg-1.0Al-1.0Zn-0.2Mn-0.5Ca Alloy Sheet Manufactured via Normal Casting and Twin Roll Casting Process (일반주조 및 쌍롤주조 공정으로 제조된 Mg-1.0Al-1.0Zn-0.2Mn-0.5Ca 합금 판재의 열처리에 따른 미세조직 및 기계적 특성 변화)

  • Dong Hwan Eom;No Jin Park
    • Journal of the Korean Society for Heat Treatment
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    • v.36 no.6
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    • pp.359-366
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    • 2023
  • Changes in microstructure and mechanical properties of Mg-1.0Al-1.0Zn-0.2Mn-0.5Ca (AZMX1100) alloy sheet manufactured by normal casting and twin roll casting process, were studied according to process and heat treatment. Non-uniform microstructure was observed in the initial sheet produced through both processes, and in particular, tilted dendrites and shifted central segregation were observed in the twin roll casting sheet. It was homogenized through hot rolling and heat treatment, and heat treated at 350℃ and 400℃ to compare the effect of heat treatment temperature. Both sheets were homogenized by the hot rolling process, and the grain size increased as the heat treatment temperature and time increased. It was confirmed that the grain size, deviation, and distribution of the second phase were finer and more homogenized in the TRC sheet. Accordingly, mechanical properties such as hardness, formability, and tensile strength also showed better values. However, unlike other previously reported AZMX alloy systems, it showed low formability (Erichsen value), which was judged by the influence of Al2Ca present in the microstructure.

The Electrochemical Chlorination for Marine Plankton Community Disinfection (해양 플랑크톤 군집의 전기분해 염소소독 효과)

  • Kang, Jung-Hoon;Shin, Kyoung-Soon;Hyun, Bong-Gil;Jang, Min-Chul;Kim, Eun-Chan;Chang, Man
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.10 no.3
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    • pp.127-137
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    • 2007
  • To confirm whether or not the Electrochemical Disinfection System (EDS) meet with the D-2 regulation established by IMO (International Maritime Organization), the biological treatment efficacy of the EDS was assessed using three groups of natural marine plankton (bacteria, $10-50\;{\mu}m$ and $>50\;{\mu}m$ sized organisms). Influent water was passed through the EDS under the flow velocity ($23.8\;m^3/hr$) and test design was consisted of control (no treatment) and experimental (10 ppm and 30 ppm) condition for total residual chlorine (TRC). And the biological condition of the influent water followed the standards established by the guidelines for the approval of ballast water management systems. The disinfection efficacy of the $10-50\;{\mu}m$ sized organisms (phytoplankton) was assessed by three kinds of measurements using photomicroscope, epifluorescence microscope and fluorometer (fumer Designs 10-AU). After being passed through the EDS, all motile phytoplankton lost their motility under photomicroscope, the colour of chlorophyll fluorescence fumed from red into green under epifluorescence, and the high chlorophyll fluorescence (Expt. 1: 6.95, Expt. 2: 7.11) detected by fluorometer decreased into value not detected. These results indicated phytoplankton community was totally killed after electrochemical disinfection treatment. Survivorship of the larger organisms than $50\;{\mu}m$ was determined based on the appendage's movement under a stereomicroscope. Natural assemblage collected from ambient seawater was killed shortly after being passed through the EDS, whereas some Artemia remained alive. However, no live Artemia was found after 24 hour further exposure to each TRC concentration (10 and 30 ppm) under darkness. After electrochemical treatment, the target bacteria such as aerobes, coliform and Escherichia coli were completely killed on the basis of CFU (colony forming unit) on Petrifilm plate ($3\;M^{TM}$) after 48 hr incubation. Moreover, no regrowth was found in the three groups of plankton during five days under additional exposure to the treated water. These results indicated that the disinfection efficiency of the EDS on the three groups of plankton satisfy D-2 regulation.

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Enhancement of Lycopene Production in Escherichia coli by Optimization of the Lycopene Synthetic Pathway

  • KANG MIN-JUNG;YOON SANG-HWAL;LEE YOUNG-MI;LEE SOOK-HEE;KIM JU-EUN;JUNG KYUNG-HWA;SHIN YONG-CHUL;KIM SEON-WON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.880-886
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    • 2005
  • Using carotenoid genes of Erwinia herbicola, metabolic engineering was carried out for lycopene production with the pAC-LYCO4 plasmid, which was composed of a chromosomal DNA fragment of E. herbicola containing the crtE, crtB, and crtI genes under the control of the tetracycline promoter and the ipi gene of Haematococcus pluvialis with the trc promoter. Plasmid pAC-LYCm4 was constructed for efficient expression of the four exogenous genes using a strong RBS sequence and the same tetracycline promoter. The optimized expression construct of pAC-LYCm4 increased Iycopene production three times as compared with pAC-LYCO4. pAC-LYCm5 containing ispA behind the four exogenous genes was constructed. There was no significant difference in Iycopene production and cell growth between pAC-LYCm4 and pAC-LYCm5. FPP synthase encoded by ispA was not rate-limiting for Iycopene production. Each gene of crtE, crtB, crtI, and ipi was overexpressed, using pBAD-crtE, pBAD-crtIB, and pBAD-ipiHPI, in addition to their expression from pAC-LYCm4. However, there was no increase oflycopene production with the additional overexpression of each exogenous gene. The four exogenous genes appeared to be not rate-limiting in cells harboring pAC-LYCm4. When pDdxs, pBAD24 containing dxs, was introduced into cells harboring lycopene synthetic plasmids, lycopene production of pAC-LYCO4, pAC-LYCm4, and pAC-LYCm5 was increased by 4.7-, 2.2-, and 2.2-fold, respectively. Lycopene production of pBAD-DXm4 containing crtE, crtB, crtI, ipi, and dxs was 5.2 mg/g dry cell weight with $0.2\%$ arabinose, which was 8.7-fold higher than that of the initial strain with pAC-LYC04. Therefore, the present study showed that proper regulation of a metabolically engineered pathway is important for Iycopene production.

Cloning and Expression of Indole Oxygenase Gene Derived from Rhodococcus sp. RHA1 (Rhodococcus sp. RHA1 유래의 Indole Oxygenase의 클로닝 및 발현)

  • Kang, Mi-Suk;Lee, Jin-Ho
    • Microbiology and Biotechnology Letters
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    • v.37 no.3
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    • pp.197-203
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    • 2009
  • An indole oxygenase originated from Rhodococcus sp. RHA1 was cloned into the expression vector, pTrc99A, in Escherichia coli, and designated pTCAN1. The pTCAN2 was constructed from pTCAN1 by the deletion of $lacI^q$ for the constitutive expression of indole oxygenase without adding IPTG in the medium. The complete open reading frame of indole oxygenase was 1,224 bp long, which encodes a protein of 407amino acids. Crude extracts of E. coli $DH5{\alpha}$/pTCAN1 and pTCAN2, respectively, were prepared and subjected to SDS-PAGE analysis. A band corresponding to molecular mass of about 43 kDa was appeared and this result correlated with the predicted molecular mass of cloned indole oxygenase. The E. coli harboring pTCAN1 and pTCAN2, respectively, showed blue color colony in LB plate. The pigment showing blue color was prepared from E. coli $DH5{\alpha}$/pTCAN2, and identified as indigo by experiments using spectrophotometer, HPLC, and TLC. The indigo-forming activity of indole oxygenases from the whole cell of E. coli $DH5{\alpha}/pTCAN1$ cultured at LB medium added 1mM of IPTG and that of E. coli/pTCAN2 showed about 1.75nmol/min/mg DCW (dry cell weight) and 3.85 nmol/min/mg DCW, respectively. Also, the E. coli $DH5{\alpha}$/pTCAN2 produced about $236{\mu}M$ of indigo after 48 hours incubation in TB medium supplemented with 2.5 mM of tryptophan.

The U.S. Legal System in Telecommunication Standardization (미국의 정보통신 표준화 법체계 연구)

  • Sohn, Hong;Park, Ki-Shik
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2000.05a
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    • pp.50-55
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    • 2000
  • The United States has had the superiority in the global trading market, and focused on the deregulation, decentralization, and often competitiveness. Also, it has adhered to pluralistic and uncoordinated systems for its various standard related activities. But with the WTO TBT entering into force late in 1990s, international standards have become more important in the global telecommunication market. So it has been recognized that the progressive standard activity would lead to keep the superiority of the nation in global telecommunication market. Specially, as the EU has been most active in building an agreed-upon technical standards among Its members, the US has faced with a serious problem that it has lack of agreed-upon infrastructure for standards. Hence, to keep the leadership in international telecommunications market, now it has been focusing on the national approach to standardization activities through the governmental support. For the implementation of above purposes, it amended 2 Acts. One is the Telecommunication Act of 1996. The other is NTTAA(National Technology Transfer and Advancement Art) of 1996, which was enacted according to the 1995 report "Standards, Conformity Assessment, and Trade into the 21 Century" by )TRC(National Research Council). In this paper, we analyse the US legal system in telecommunication standardization field including above arts and their Implementing plans. And we suggest the need for the active system of government in our telecommunication standardization.

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