• Title/Summary/Keyword: direct synthesis

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Enhanced In Vitro Protein Synthesis Through Optimal Design of PCR Primers

  • Ahn Jin-Ho;Son Jeong-Mi;Hwang Mi-Yeon;Kim Tae-Wan;Park Chang-Kil;Choi Cha-Yong;Kim Dong-Myung
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.355-359
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    • 2006
  • The functional stability of mRNA is one of the crucial factors affecting the efficiency of in vitro translation. As the rapid degradation of mRNA in the cell extract (S30 extract) causes early termination of the translational reactions, extending the mRNA half-life will improve the productivity of the in vitro protein synthesis. Thus, a simple PCR-based method is introduced to increase the stability of mRNA in an S30 extract. The target genes are PCR-amplified with primers designed to make the ends of the transcribed mRNA molecule anneal to each other. When compared with normal mRNA, the mRNA with the annealing sequences resulted in an approximately 2-fold increase of protein synthesis in an in vitro translation reaction. In addition, sequential transcription and translation reactions in a single tube enabled direct protein expression from the PCR-amplified genes without any separate purification of the mRNA.

Physiological and Genetic Factors Controlling Streptomyces Regulatory Gene Expression Involved in Antibiotic Biosynthesis

  • Kim Eung Su
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.68-72
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    • 2002
  • While the biosynthetic gene cluster encoding the pigmented antibiotic actinorhodin is present in the two closely related bacterial species, Streptomyces lividans and Streptomyces coelicolor, it normally is expressed only in S. coelicolor---generating the deep blue colonies responsible for the S. coelicolor name. However, multiple copies of the afsR2 gene, which activates actinorhodin synthesis, result in the ability of S. lividansto also synthesize large amounts of actinorhodin. Here we report that the phenotypic property that historicially distinguishes these two Streptomycesspecies is determined conditionally by the carbon source used for culture. Whereas growth on glucose repressed actinorhodin production in S. lividans, culture on solid media containing glycerol as the sole carbon source dramatically increased the expression of afsR2 mRNA---leading to extensive actinorhodin synthesis by S. lividansand obliterating its phenotypic distinction from S. coelicolor. afsR2 transcription under these conditions was developmentally regulated, rising sharply at the time of aerial mycelium formation and coinciding temporally with the onset of actinorhodin production. Our results, which identify media-dependent parallel pathways that regulate actinorhodin synthesis in S. lividans, demonstrate carbon source control of actinorhodin production through the regulation of afsR2 mRNA synthesis. The nucleotide sequences of afsR2 revealed two putative important domains; the domain containing direct repeats in the middle and the domain homologous to sigma factor sequence in the C-terminal end. In this work, we constructed various sized afsR2-derivatives and compared the actinorhodin stimulating effects in S. lividans TK21. The experimental data indicate that the domain homologous to sigma factor sequence in the C-terminal end of afsR2 plays a critical role as an antibiotic stimulating function. In addition, we also observed that the single copy integration of afsR2 regulatory gene into S. lividans TK21 chromosome significantly activates antibiotic overproduction.

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Exploration of growth mechanism for layer controllable graphene on copper

  • Song, Woo-Seok;Kim, Yoo-Seok;Kim, Soo-Youn;Kim, Sung-Hwan;Jung, Dae-Sung;Jun, Woo-Sung;Jeon, Cheol-Ho;Park, Chong-Yun
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.490-490
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    • 2011
  • Graphene, hexagonal network of carbon atoms forming a one-atom thick planar sheet, has been emerged as a fascinating material for future nanoelectronics. Huge attention has been captured by its extraordinary electronic properties, such as bipolar conductance, half integer quantum Hall effect at room temperature, ballistic transport over ${\sim}0.4{\mu}m$ length and extremely high carrier mobility at room temperature. Several approaches have been developed to produce graphene, such as micromechanical cleavage of highly ordered pyrolytic graphite using adhesive tape, chemical reduction of exfoliated graphite oxide, epitaxial growth of graphene on SiC and single crystalline metal substrate, and chemical vapor deposition (CVD) synthesis. In particular, direct synthesis of graphene using metal catalytic substrate in CVD process provides a new way to large-scale production of graphene film for realization of graphene-based electronics. In this method, metal catalytic substrates including Ni and Cu have been used for CVD synthesis of graphene. There are two proposed mechanism of graphene synthesis: carbon diffusion and precipitation for graphene synthesized on Ni, and surface adsorption for graphene synthesized on Cu, namely, self-limiting growth mechanism, which can be divided by difference of carbon solubility of the metals. Here we present that large area, uniform, and layer controllable graphene synthesized on Cu catalytic substrate is achieved by acetylene-assisted CVD. The number of graphene layer can be simply controlled by adjusting acetylene injection time, verified by Raman spectroscopy. Structural features and full details of mechanism for the growth of layer controllable graphene on Cu were systematically explored by transmission electron microscopy, atomic force microscopy, and secondary ion mass spectroscopy.

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Identification of a Domain in Yeast Chitin Synthase 3 Required for Biogenesis of Chitin Ring, But Not Cellular Chitin Synthesis

  • Park Hyun-Sook;Park Mee-Hyun;Kim Chi-Hwa;Woo Jeeun;Lee Jee-Yeon;Kim Sung-Uk;Choi Wonja
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2000.10a
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    • pp.39-45
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    • 2000
  • It hab been proposed that CHS3-mediated chitin synthesis during the vegitative cell cycle is regulated by CHS4. To investigate direct protein-protein interaction between their coding products, we used yeast two hybrid system and found that a domain of Chs3p was responsible for interaction with Chs4p. This domain, termed MIRC3-4 (maximum interacting region of chs3p with chs4p), spans from 647 to 700 residues. It is well conserved among CHS3 homologs of various fungi such as Candida albicans, Emericella nidulans, Neurospora crassa, Magnaporthe grisea, Ustilago maydis, Glomus versiforme, Exophiala dermatitidis, Rhizopus microsporus. A series of mutaion in the MIRC3-4 resulted in no appearance of chitin ring at the early G 1 phase but did not affect chitin synthesis in the cell wall after cytokinesis. Absence of chitin ring could be caused either by delocalization of Chs3p to the septum or by improper interaction with Chs4p. To discriminate those two, not mutually exclusive, alternatives, mutants cells were immunostained with Chs3p-specific antibody. Some exhibited localization of chs3p to the septum, while others failed. These results indicate that simultaneous localization and activation Chs3p by Chs4p is required for chitin ring synthesis.

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Optimization of KOGAS DME Process From Demonstration Long-Term Test (KOGAS DME 공정의 실증 시험을 통한 최적화 기술개발)

  • Chung, Jongtae;Cho, Wonjun;Baek, Youngsoon;Lee, Changha
    • Journal of Hydrogen and New Energy
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    • v.23 no.5
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    • pp.559-571
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    • 2012
  • Dimethyl ether (DME) is a new clean fuel as an environmentally-benign energy resource. DME can be manufactured from various energy sources including natural gas, coal, and biomass. In addition to its environmentally friendly properties, DME has similar characteristics to those of LPG. The aim of this article is to represent the development of new DME process with KOGAS's own technologies. KOGAS has investigated and developed new innovative DME synthesis process from synthesis gas in gaseous phase fixed bed reactor. DME has been traditionally produced by the dehydration of methanol which is produced from syngas, a product of natural gas reforming. This traditional process is thus called the two-step method of preparing DME. However, DME can also be manufactured directly from syngas (single-step). The single-step method needs only one reactor for the synthesis of DME, instead of two for the two-step process. It can also alleviate the thermodynamic limitations associated with the synthesis of methanol, by converting the produced methanol into DME, thereby potentially enhancing the overall conversion of syngas into DME. KOGAS had launched the 10 ton/day DME demonstration plant project in 2004 at Incheon KOGAS LNG terminal. In the mid of 2008, KOGAS had finished the construction of this plant and has successively finished the demonstration plant operation. And since 2008, we have established the basic design of commercial plant which can produce 3,000 ton/day DME.

Direct Methanol Synthesis by Partial Oxidation of Methane (메탄의 부분산화에 의한 메탄올 직접 합성)

  • Kim, Young-Kook;Lee, Kwang-Hyeok;Hahm, Hyun-Sik
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.4
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    • pp.649-655
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    • 2013
  • Methanol was directly produced by the partial oxidation of methane with perovskite and mixed oxide catalysts. Perovskite ($ABO_3$) catalysts were prepared by the malic acid method with changing A and B site components. Three-component mixed oxide catalysts that have Mo and Bi as a main component were prepared by the co-precipitation method. Among the perovskite catalysts, $SrCrO_3$ showed the highest methanol selectivity of 11% at $400^{\circ}C$. For the three-component mixed oxide catalysts, there were no remarkable changes in methane conversion. Among the mixed oxide catalysts, Mo-Bi-Cr mixed oxide catalyst showed the highest methanol selectivity of 15.3% at $400^{\circ}C$. The catalytic activity and methanol selectivity of the three-component mixed oxide catalysts were directly proportional to the surface area of the catalysts.

Preliminary Feasibility Study for Commercial DME Plant Project (DME 상용화 플랜트 예비 타당성 조사)

  • Mo, Younggi;Kang, Minseo;Song, Taekyoung;Baek, Youngsoon;Cho, Wonjun
    • Journal of Hydrogen and New Energy
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    • v.25 no.2
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    • pp.173-182
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    • 2014
  • Dimethyl ether (DME) is a new clean fuel as an environmentally-being energy resources. DME has similar characteristics to those of LPG and can be substituted Diesel fuel. KOGAS has investigated and developed new innovative DME synthesis process from synthesis gas with KOGAS's own technologies. KOGAS had finished the construction of 10ton/day DME demonstration plant in 2008, we have established the basic design of commercial plant which can produce 3,000ton/day DME. Specifically, an economic model for a commercial DME project will be presented. It accounts for all the major cost factors that are considered in a commercial scale project as the model input for performing cash flow analysis, after which key economic indicators are produced including the internal rate of return (IRR), net present value (NPV). Sensitivity analysis is performed to identify dominant cost factors to the project economics and quantify their impact. The inputs to the economic analysis will be based on representative cost factors from the commercial-scale design of KOGAS' direct DME process supplemented by literature data. Case study results will be presented based on recent commercialization projects.

Controlled Synthesis of Single-Walled Carbon Nanotubes

  • Park, Chong-Yun
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.2-2
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    • 2011
  • Single-walled carbon nanotubes (SWNTs) have been considered as a promising candidate for nextgeneration electronics due to its extraordinary electrical properties associated with one-dimensional structure. Since diversity in electronic structure depends on geometrical features, the major concern has been focused on obtaining the diameter, chirality, and density controlled SWNTs. Despite huge efforts, the controlled synthesis of SWNTs has not been achieved. There have been various approaches to synthesize controlled SWNTs by preparation of homogeneously sized catalyst because the SWNTs diameter highly depends on catalyst nanoparticles size. In this study, geometrically controlled SWNTs were synthesized using designed catalytic layers: (a) morphologically modified Al2O3 supporting layer (Fe/Al2O3/Si), (b) Mo capping layer (Mo/Fe/Al/Si), and (c) heat-driven diffusion and subsequent evaporation process of Fe catalytic nanoparticles (Al2O3/Fe/Al2O3/Si). These results clearly revealed that (a) the grain diameter and RMS roughness of Al2O3 supporting layer play a key role as a diffusion barrier for obtaining Fe nanoparticles with a uniform and small size, (b) a density and diameter of SWNTs can be simultaneously controlled by adjusting a thickness of Mo capping layer on Fe catalytic layer, and (c) SWNTs diameter was successfully controlled within a few A scale even with its fine distribution. This precise control results in bandgap manipulation of the semiconducting SWNTs, determined by direct comparison of Raman spectra and theory of extended tight binding Kataura plot. We suggest that these results provide a simple and possible way for the direct growth of diameter, density, and bandgap controlled SWNTs by precise controlling the formation of catalytic films, which will be in demand for future electronic applications.

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Chiral Mesoporous Silica for Asymmetric Metal-free Catalysis: Enhancement of Chirality thorough Confinement Space by Plug Effect

  • Jeong, Eun-Yeong;Im, Cheong-Rae;Park, Sang-Eon
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.08a
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    • pp.199-199
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    • 2011
  • The addition of a carbanion to ${\yen}{\acute{a}}{\yen}{\hat{a}}$-unsaturated carbonyl compounds is of importance in the C-C bond formation reactions for modern pharmaceuticals and organic synthesis. Recently, heterogeneous asymmetric catalysis became more attractive area of research because of the easy recovery and separation of the catalyst from the reaction system. Most of synthetic methods for heterogeneous catalysts were grafting or immobilization of homogeneous catalyst onto the solid supports. Trans-1,2-Diaminocyclohexane(DACH) and L-proline ligands have been enormously used as chiral ligands in several catalytic transformation under homogenous conditions. Our group prepared l-proline functionalized mesoporous silica was synthesized under acidic condition using a poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer template (EO20PO70EO20, Pluronic P-123, BASF). Furthermore, we successfully directly synthesized trans-1,2 diaminocyclohexane functionalized mesoporous silica by using microwave method. The direct functionalization of chiral ligand into the framework of mesoporous materials is expected to be useful for the heterogeneous asymmetric catalysis. So, we adopt the direct synthesis of chiral ligand functionalized mesoporous silica by using thermal and microwave irradiation. Then, chiral ligand functionalized mesoporous silicas were applied to enantioselective asymmetric catalytic reactions.

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A Method for Suppression of Active Metal Leaching during the Direct Synthesis of H2O2 by Using Polyelectrolyte Multilayers (고분자 전해질 다층박막을 이용한 과산화수소 직접제조 반응 중 활성금속 용출 억제 방법)

  • Chung, Young-Min
    • Korean Chemical Engineering Research
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    • v.53 no.2
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    • pp.262-268
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    • 2015
  • In this study, two types of catalysts were prepared via conventional metal supporting method and encapsulation of metal nanoparticles in the polyelectrolyte multilayers constructed on support. The resulting catalysts were applied to the direct synthesis of hydrogen peroxide, and the effect of catalyst preparation method on the catalyst life as well as hydrogen peroxide productivity was investigated. The catalytic activity was strongly dependent upon the acid strength of support regardless of the catalyst preparation methods and HBEA (SAR=25) with strong acidity was superior to other supports to promote the reaction. In the case of metal supported catalyst, while hydrogen peroxide productivity was higher than that of polyelectrolyte multilayered counterpart, the reaction performance was sharply decreased during catalyst recycling due to the metal leaching. On the other hand, construction of polyelectrolyte multilayers on support weakened the influence of acid support on the reaction medium and therefore resulted in the decrease of catalytic activity and the increase of hydrogen peroxide decomposition as well. It is noted, however, that the catalytic activity was maintained after 5 recycles, which suggests that the introduction of polyelectrolyte multilayers on the support is very effective to suppress the unfavorable metal leaching phenomenon during a reaction.