• Title/Summary/Keyword: Multifunctional System

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Genenation of structural diversity in polyketides by combinatorial biosynthesis of polyketides: Part I. Generation of multiple bioactive macrolides by hybrid modular polyketide synthases in Streptomyces venezuelae, Part II. Production of novel rifamycins by combinatorial biosynthesis

  • Yoon, Yeo-Joon
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2002.10a
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    • pp.18-25
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    • 2002
  • The pikromycin biosynthetic system in Streptomyces venezuleae is unique for its ability to produce two groups of antibiotics that include the 12-membered ring macrolides methymycin and neomethymycin, and the 14-membered ring macrolides narbomycin and pikromycin. The metabolic pathway also contains two post polyketide-modification enzymes, a glycosyltransferase and P450 hydroxylase that have unusually broad substrate specificities. In order to explore further the substrate flexibility of these enzymes a series of hybrid polyketide synthases were constructed and their metabolic products characterized. The plasmid-based replacement of the multifunctional protein subunits of the pikromycin PKS in S. venezuelae by the corresponding subunits from heterologous modular PKSs resulted in recombinant strains that produce both 12- and 14-membered ring macrolactones with predicted structural alterations. In all cases, novel macrolactones were produced and further modified by the DesVII glycosyltransferase and PikC hydroxylase leading to biologically active macrolide structures. These results demonstrate that hybrid PKSs in S. venezuelae can produce a multiplicity of new macrolactones that are modified further by the highly flexible DesVII glycosyltransferase and PikC hydroxylase tailoring enzymes. This work demonstrates the unique capacity of the S. venezuelae pikromycin pathway to expand the toolbox of combinatorial biosynthesis and to accelerate the creation of novel biologically active natural products. The polyketide backbone of rifamycin B is assembled through successive condensation and ${\beta}$-carbonyl processing of the extender units by the modular rifamycin PKS. The eighth module, in the RifD protein, contains nonfunctional DH domain and functional KR domain, which specify the reduction of the ${\beta}$-carbonyl group resulting in the C-21 bydroxyl of rifamycin B. A four amino acid substitution and one amino acid deletion were introduced in the putative NADPH binding motif in the proposed KR domain encoded by rifD. This strategy of mutation was based on the amino acid sequences of the corresponding motif of the KR domain of module 3 in the RifA protein, which is believed dysfunctional, so as to introduce a minimum alteration and retain the reading frame intact, yet ensure loss of function. The resulting strain produces linear polyketides, from tetraketide to octaketide, which are also produced by a rifD disrupted mutant as a consequence of premature termination of polyketide assembly. Much of the structural diversity within the polyketide superfamily of natural products is due to the ability of PKSs to vary the reduction level of every other alternate carbon atom in the backbone. Thus, the ability to introduce heterologous reductive segments such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER) into modules that naturally lack these activities would increase the power of the combinatorial biosynthetic toolbox. The dehydratase domain of module 7 of the rifamycin PKS, which is predicted to be nonfunctional in view of the sequence of the apparent active site, was replaced with its functional homolog from module 7 of rapamycin-producing polyketide synthase. The resulting mutant strain behaved like a rifC disrupted mutant, i.e., it accumulated the heptaketide intermediate and its precursors. This result points out a major difficulty we have encountered with all the Amycolatopsis mediterranei strain containing hybrid polyketide synthases: all the engineered strains prepared so far accumulate a plethora of products derived from the polyketide chain assembly intermediates as major products instead of just analogs of rifamycin B or its ansamycin precursors.

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Immune-alteration Demonstrated at the Korean Vietnam War Veterans Exposed to Agent Orange (2,3,7,8-tetrachlorodibenzo-p-dioxin 노출과 관련한 인체면역기능 변화를 판단할 수 있는 지표치 개발에 관한 연구)

  • Heo, Yong;Kim, Eun-Mi;Yu, Ji-Yeon;Hong, Seung-Kwon;Jeon, Seong-Hoon;Kim, Hyoung-Ah;Cho, Dae-Hyun;Han, Soon-Young
    • Environmental Mutagens and Carcinogens
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    • v.22 no.2
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    • pp.112-124
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    • 2002
  • 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been reported to exert detrimental toxicities on various organ systems including reproductive, cardiovascular, nervous, or dermal system. Immunomodulatory effects of TCDD is thymic atrophy, downregulation of cytotoxic T or B lymphocyte differentiation and activation, which were demonstrated using experimental animals, whereas immunotoxicity in human has not been investigated well. This study was proceeded to evaluate general immunologic spectrum of the Korean Vietnam War veterans exposed to TCDD during their operation, and compare with that of the non-exposed control subjects with similar age. Regarding composition and quantity, immune cells in peripheral blood collected from the TCDD-exposed was not much different from those of the control except decreased red blood cell, hemoglobin and hematocrit level. Furthermore, plasma IgG2, G3, and G4 isotype distribution was similar between two groups, but IgG1 level was significantly lowered in the TCDD-exposed, indicating a TCDD-mediated functional alteration of B cells. Significantly enhanced level of IgE in plasma, a hallmark of dermal or respiratory allergic response, was also observed in the TCDD-exposed compared with that of the control. Elevated generation of IL-4 and IL-10 was resulted from in vitro stimulation of T cells with PMA plus ionomycin or PHA, respectively, from the TCDD-exposed in comparison to those of the control, suggesting a skewed type-2 response. In addition, the level of IFN${\gamma}$, a multifunctional cytokine for T cell-mediated immunity, was lowered in the TCDD-exposed with upregulation of tumor necrosis factor $\alpha$. The present study suggests that TCDD exposure disturbs immunohomeostasis in humans observed as an aberrant plasma IgE and IgG1 levels and dysregulation of T cell activities.

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Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice

  • Chung, Eu Jin;Hossain, Mohammad Tofajjal;Khan, Ajmal;Kim, Kyung Hyun;Jeon, Che Ok;Chung, Young Ryun
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.152-164
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    • 2015
  • Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.

Effect of Wrapping Treatment on the Dispersion of MWNT in CNT/ABS/SAN Composites (CNT/ABS/SAN계의 분산성에 미치는 MWNT Wrapping 전처리 효과)

  • Kim, Sung Tae;Park, Hae Youn;No, Tae Kyeong;Kang, Dong Gug;Jeon, Il Ryeon;Seo, Kwan Ho
    • Applied Chemistry for Engineering
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    • v.23 no.4
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    • pp.372-376
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    • 2012
  • Carbon nanotubes (CNT) are considered as one of ideal nano-fillers in the field of composites with their excellent electrical, mechanical, and thermal properties. Therefore CNT composites are increasingly used in fabricating conductive materials, structural materials with high strength and low weight, and multifunctional materials. The main problem of the CNT composites is difficulty in the dispersion of CNT in the polymer matrix. In this study multi-walled carbon nanotubes (MWNT) were pretreated by the physical process utilizing a wrapping method. After the pretreatment polymer/MWNT nanocomposites were prepared by melt processing. The effect of functionalization MWNT by wrapping with styrene acrylonitrile (SAN) on the mechanical and electrical properties of acrylonitrile butadiene styrene resin (ABS)/MWNT composites was studied by comparing the properties of ABS mixed with the neat MWNT. Electrical and mechanical properties of ABS/MWNT nanocomposites were studied as a function of the functionalization and content of MWNT. The tensile strength of the ABS/MWNT nanocomposites increased, but the impact strength decreased. The polymer wrapping in ABS system has little effect on the improvement of electrical properties.

Trends of Supply and Demand of Aggregate in Korea (I) (국내 골재자원의 수급현황(I))

  • Hong, Sei Sun;Kim, Ju Yong;Lee, Jin Young
    • The Journal of the Petrological Society of Korea
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    • v.24 no.3
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    • pp.253-272
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    • 2015
  • The necessity of aggregate resources in Korea has been increased with a rapid economic growth since the 1990s. Construction activities have been increased drastically for last two decades. Such economic activities are much concentrated at big cities in Korea, and recently new neighbouring cities, such as multifunctional administrative city, enterprise city and innocity, are under construction at the surroundings of the cities. This new urbanization asked for an appropriate supply of raw construction materials such as cement, sands and gravels. The aim of this study is to understand and discuss the analysis and prospect of supply and demand of domestic sand and gravel. On the early years, 1990s, about 25% of the total consumption of these sands and gravels comes from riverine deposits; 20% to 25% from marine sands, 40% to 45% from forest rocks and the rest 5% to 15% from old fluvial deposits and crushed rocks. But nowaday the river aggregate, including both sands and gravels of a present river channel and those of the old fluviatile system are decreasing, while the crushed and forest aggregates and marine sand are now the main source of infrastructure resources. Thus it is increasingly necessary to investigate the forest and crushed aggregate resources potential in Korea where a current analysis indicate that the supply of riverine aggregates gradually decreases.

Dashboard Design for Evidence-based Policymaking of Sejong City Government (세종시 데이터 증거기반 정책수립을 위한 대시보드 디자인에 관한 연구)

  • Park, Jin-A;An, Se-Yun
    • The Journal of the Korea Contents Association
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    • v.19 no.12
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    • pp.173-183
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    • 2019
  • Sejong, Korea's special multifunctional administrative city, was created as a national project to relocated government ministries, the aim being to pursue more balanced regional economic development and boost national competitiveness. During the second phase development will focus on mitigating the challenges raised due to the increasing population and urbanization development. All of infrastructure, apartments, houses, private buildings, commercial structures, public buildings, citizens are producing more and more complex data. To face these challenges, Sejong city governments and policy maker recognizes the opportunity to ensure more enriched lives for citizen with data-driven city management, and effectively exploring how to use existing data to improve policy services and a more sustainable economic policy to enhance sustainable city management. As a city government is a complex decision making system, the analysis of astounding increase in city dada is valuable to gain insight in the affecting traffic flow. To support the requirement specification and management of government policy making, the graphic representation of information and data should be provide a different approach in the intuitive way. With in context, this paper outlines the design of interactive, web-based dashboard which provides data visualization regarding better policy making and risk management.

Study on the Improved Abrasion Resistance of Polycarbonate Substrate by UV-curable Organic/Inorganic Hybrid Coatings (자외선 경화형 유기/무기 복합코팅에 의한 폴리카보네이트의 내마모성 향상 연구)

  • 윤석은;우희권;김동표
    • Polymer(Korea)
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    • v.24 no.3
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    • pp.389-398
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    • 2000
  • Transparent, abrasion resistant coatings with 4~13 ${\mu}{\textrm}{m}$ thickness were prepared by spin-coating on polycarbonates with organic/inorganic hybrid solutions, followed by UV curing and heat treatment at 12$0^{\circ}C$ for 12 hours. The coating solutions were composed of inorganic phase and organic phase in 0:100, 20:80, 30:70, 50:50, 80:20 wt% ratios, respectively, mixed with photoinitiator, senaitizer and surfactant. The inorganic phase was formed by sol-gel reaction of TEOS and silane coupling agent MPTMS in 1 : 2 or 2 : 1 molar ratios, the organic phase consisted of difunctional urethane acrylate oligomeric resin, multifunctional acrylate TMPTA and HDDA in 4 : 3 : 3 wt% ratio. The coating systems were investigated by FT-IR, $^{29}$ Si-NMR spectra. In addition, TGA/DSC for thermal analysis and SEM, AFM observation for coated surface were examined. Gererally, the homogeneity of phases, the surface smoothness of coating and abrasion resistance were improved with the higher content of inorganic component. Namely, coating system with below 10 $\AA$ surface roughness and T$_{g}$ of 15$0^{\circ}C$ showed only 10% decrease in light transmittance after abrasion test, whereas uncoated polycarbonate substrate exhibited 46% decrease..

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A Study on Evaluating Tourism Value and Presenting Development Strategies for the Saemangeum Project (새만금지역의 관광자원 가치와 개발방안에 관한 연구)

  • Kim, Hak Yong
    • Journal of Industrial Convergence
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    • v.17 no.2
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    • pp.9-14
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    • 2019
  • This study was carried out to investigate how the basic plan of the Saemangeum development project that has been developed by the government since 1991 has been changed and to present a new development strategy by analyzing the future value and tourism resource value of Saemangeum development. The research methodology is based on the in - depth analysis methodology of the time series research materials and press releases published by the Saemangeum Development Administration, which oversees the development of Saemangeum. The government announced a plan to significantly reduce agricultural land and increase the proportion of non-agricultural land to 70% in order to create a multifunctional complex. After that, the government revised the basic plan of Saemangeum in 2014 and changed the plan to tailor the investment demand in order to cultivate investment demand, and introduced the special economic cooperation system for global economic cooperation. Therefore, the purpose of this paper is to analyze the process of Saemangeum development to date and to evaluate the value of Saemangeum development as a tourism resource, centered on the Master Plan of the Saemangeum Development Project changed by the government. Based on the results of this analysis, I would like to present the vision of Saemangeum tourism development considering Saemangeum 's environment and characteristics of the surrounding scenery by referring to trends and similar cases of international tourism.

Inhibitory Effort of the N-terminal GST on the Tautomerase Activity of Macrophage Migration Inhibitory Factor (GST 융합 시스템에서 나타나는 macrophage migration inhibitory factor의 tautomerase 활성 저해에 관한 연구)

  • Kim Sang-Soo;Kim Kyung-Hee;Park Hyo-Jin;Hur Eun-hye;Rhim Hyangshuk
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.961-967
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    • 2005
  • Macrophage migration inhibitory fartor (MIF), known as a cytokine, is a multifunctional protein that is ubiquitously expressed in a variety of cells and tissues; however, enzymatic function of MIF still remains elusive in cells. In this study, we assessed details of the tautomerase activity of MIF. We established rapid purification condition for MIF by using pGEX system and compared the L-dopachrome tautomerase activity of GST-MIF, tMIF, and MIF. The results show that GST (glutathione S-transferase)-epitope tag or N-terminal amino acids flanking the essential $P^{2}$ almost completely abrogated L-dopachrome tautomerase activity of MIF. Subsequently, to determine whether the N-terminal tags have effects on oligomerization of MIF, protein cross-linking products were analyzed on $15\%$ SDS-PACE. The result demonstrates that N-terminal tags are dispensable for the formation of MIF's homooligomers. Thus, the results imply that exposure of If containing hydrophobic pocket in the active site is critical for L-dopachrome tautomerase activity of MIF. In addition, our study suggest that the MIF's tautomerase activity might be influenced by interacting with cellular partners.

Effects of Rhizoma Coptidis on Cellular Activity and IL-6 Production of LPS-treated Periodontal Ligament Cells (황련이 Lipopolysaccharide를 처리한 치주인대세포의 세포활성 및 IL-6 생산에 미치는 영향)

  • Song, Ki-Bum;Kong, Young-Hwan;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.3
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    • pp.641-654
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    • 1996
  • In infectious disease, invasion of host tissue by bacteria or their products frequently induces a wide variety of inflammatory and immunopathologic reaction. Evidence indicates that cytokines are involved in the initiation and progression of chronic inflammatory diseases, such as periodontitis. Interleukin-6, which is a multifunctional cytokine, has important roles in acute and chronic inflammation and may also be implicated in bone resorption. Periodontal diseases are characterized by chronic inflammation of the periodontium with alveolar bone resoption. A principal driving force behind this response appears to lie in the immune system's response to bacteria. Many of the cell components which have been shown to function as virulence factors in gram-negative bacteria are associated with the bacterial surface. Of these, lipopolysaccharide has been characterized as one that mediates a number of biological activities which can lead to the destruction of host tissue. Non-steroidal antiinflammatory drug is used for reduce inflammation, and most of NSAIDs inhibit prostaglandine $E_2$ production, but it is shown that $PGE_2$ production is stimulated by IL-1 in recent study. So, the influence of other cytokines except $PGE_2$ on periodontium can not be avoided. Therefore, new antiinflammatory drug is needed. Rhizoma coptidis is used in oriental medicine for anti-inflammation and antiseptics. In this present study, we examined the IL-6 release in periodontal ligament cells treated with the lipopolysaccharide, and also the effect of rhizoma coptidis on cellular activity and IL-6 production of periodontal ligament cells. To evaluate the effect of rhizoma coptidis on cellular activity, the cells were seeded at a cell density of $1{\times}10^4$ cells/well in 24-well culture plates. After one day incubation, 1-6, 10-9 and 10-12 g/ml of rhizoma coptidis and 5, $10{\mu}g/ml$ of LPS were added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay were carried out. To evaluate the effect of rhizoma coptidis on IL-6 production, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates. After one day incubation, 10-9 g/ml of rhizoma coptidis and 5, $10{\mu}g/ml$ of LPS were added to the each well and incubated for 3, 6, 12 and 24 hours. Then, amounts of IL-6 production is measured by IL-6 ELISA kit used. The results were as follows : 1. Rhizoma coptidisrbelow to ($10^{-6}g/ml$) significantly increaed cellular activity of periodontal ligament cells than control. 2. Rhizoma coptidist ($10^{-9}g/ml$) significantly increased cellular activity of LPS($5{\mu}g/ml$)-treated periodontal ligament cells than control. 3. LPS(5 and $10{\mu}g/ml$) significantly increased IL-6 production of periodontal ligament cells than control. 4. Rhizoma coptidis($10^{-9}g/ml$) decreased IL-6 production of LPS ($5{\mu}g/ml$)-treated periodontal.ligarnent cells than LPS only tested group. These findings suggest that stimulation of the IL-6 release of periodontal ligament cells by LPS may have a role in the progression of inflammation and alveolar bone resoption in periodontal disease, and that inhibition of the IL-6 release of cells and stimulation of cellular activity by rhizoma coptidis may help the periodontal regeneration.

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