• Title/Summary/Keyword: Biological synthesis

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Zeolite-Mediated Cation Exchange Enhances the Stability of mRNA during Cell-Free Protein Synthesis

  • Kim, You-Eil;Kim, Dong-Myung;Choi, Cha-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.258-261
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    • 2006
  • The addition of zeolite particles enhances the stability of mRNA molecules in a cell-free protein synthesis system. When $20{\mu}g/{\mu}L$ of zeolite (Y5.4) is added to a reaction mixture of cell-free protein synthesis, a substantial increase in protein synthesis is observed. The stabilizing effect of zeolite is most dearly observed in an in vitro translation reaction directed by purified mRNA, as opposed to a coupled transcription and translation reaction. Upon the addition of zeolite in the in vitro translation reaction, the life span of the mRNA molecules is substantially extended, leading to an 80% increase in protein synthesis. The effect of zeolite upon the mRNA stability appears be strongly related to the cation exchange (potassium to sodium) reaction. Our results demonstrate the possibility of modifying this biological process using heterogeneous, non-biological substances in a cell-free protein synthesis system.

Combinatorial Solid Phase Peptide Synthesis and Bioassays

  • Shin, Dong-Sik;Kim, Do-Hyun;Chung, Woo-Jae;Lee, Yoon-Sik
    • BMB Reports
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    • v.38 no.5
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    • pp.517-525
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    • 2005
  • Solid phase peptide synthesis method, which was introduced by Merrifield in 1963, has spawned the concept of combinatorial chemistry. In this review, we summarize the present technologies of solid phase peptide synthesis (SPPS) that are related to combinatorial chemistry. The conventional methods of peptide library synthesis on polymer support are parallel synthesis, split and mix synthesis and reagent mixture synthesis. Combining surface chemistry with the recent technology of microelectronic semiconductor fabrication system, the peptide microarray synthesis methods on a planar solid support are developed, which leads to spatially addressable peptide library. There are two kinds of peptide microarray synthesis methodologies: pre-synthesized peptide immobilization onto a glass or membrane substrate and in situ peptide synthesis by a photolithography or the SPOT method. This review also discusses the application of peptide libraries for high-throughput bioassays, for example, peptide ligand screening for antibody or cell signaling, enzyme substrate and inhibitor screening as well as other applications.

Bio-Inspired Green Nanoparticles: Synthesis, Mechanism, and Antibacterial Application

  • Velusamy, Palaniyandi;Kumar, Govindarajan Venkat;Jeyanthi, Venkadapathi;Das, Jayabrata;Pachaiappan, Raman
    • Toxicological Research
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    • v.32 no.2
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    • pp.95-102
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    • 2016
  • In the recent years, noble nanoparticles have attracted and emerged in the field of biology, medicine and electronics due to their incredible applications. There were several methods have been used for synthesis of nanoparticles such as toxic chemicals and high energy physical procedures. To overcome these, biological method has been used for the synthesis of various metal nanoparticles. Among the nanoparticles, silver nanoparticles (AgNPs) have received much attention in various fields, such as antimicrobial activity, therapeutics, bio-molecular detection, silver nanocoated medical devices and optical receptor. Moreover, the biological approach, in particular the usage of natural organisms has offered a reliable, simple, nontoxic and environmental friendly method. Hence, the current article is focused on the biological synthesis of silver nanoparticles and their application in the biomedical field.

Enhanced Production of Fatty Acids via Redirection of Carbon Flux in Marine Microalga Tetraselmis sp.

  • Han, Mi-Ae;Hong, Seong-Joo;Kim, Z-Hun;Cho, Byung-Kwan;Lee, Hookeun;Choi, Hyung-Kyoon;Lee, Choul-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.267-274
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    • 2018
  • Lipids in microalgae are energy-rich compounds and considered as an attractive feedstock for biodiesel production. To redirect carbon flux from competing pathways to the fatty acid synthesis pathway of Tetraselmis sp., we used three types of chemical inhibitors that can block the starch synthesis pathway or photorespiration, under nitrogen-sufficient and nitrogen-deficient conditions. The starch synthesis pathway in chloroplasts and the cytosol can be inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and 1,2-cyclohexane diamine tetraacetic acid (CDTA), respectively. Degradation of glycine into ammonia during photorespiration was blocked by aminooxyacetate (AOA) to maintain biomass concentration. Inhibition of starch synthesis pathways in the cytosol by CDTA increased fatty acid productivity by 27% under nitrogen deficiency, whereas the blocking of photorespiration in mitochondria by AOA was increased by 35% under nitrogen-sufficient conditions. The results of this study indicate that blocking starch or photorespiration pathways may redirect the carbon flux to fatty acid synthesis.

Cyclic Peptides as Therapeutic Agents and Biochemical Tools

  • Joo, Sang-Hoon
    • Biomolecules & Therapeutics
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    • v.20 no.1
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    • pp.19-26
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    • 2012
  • There are many cyclic peptides with diverse biological activities, such as antibacterial activity, immunosuppressive activity, and anti-tumor activity, and so on. Encouraged by natural cyclic peptides with biological activity, efforts have been made to develop cyclic peptides with both genetic and synthetic methods. The genetic methods include phage display, intein-based cyclic peptides, and mRNA display. The synthetic methods involve individual synthesis, parallel synthesis, as well as split-and-pool synthesis. Recent development of cyclic peptide library based on split-and-pool synthesis allows on-bead screening, in-solution screening, and microarray screening of cyclic peptides for biological activity. Cyclic peptides will be useful as receptor agonist/antagonist, RNA binding molecule, enzyme inhibitor and so on, and more cyclic peptides will emerge as therapeutic agents and biochemical tools.

An In Vitro Assay to Screen for Translation Inhibitors

  • Song, Chin-Hee;Paik, Hyoung-Rok;Seong, Chi-Nam;Choi, Sang-Ki
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1646-1649
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    • 2006
  • Protein synthesis is the ultimate outcome of gene expression which, in turn, is regulated by several translation factors. We attempted to identify substances that can inhibit the translation process in vitro when the outcome protein is luciferase. To this end, we developed a sensitive cell-free protein synthesis assay using luciferase as the reporter. The synthesis of luciferase increased proportionately as mRNA was added to a $15-{\mu}l$reaction medium in concentrations raging from 5 ng to 500 ng. The maximum amount of luciferase was synthesized when the media were incubated at $25^{\circ}C$ for 40 min. The concentration of each compound that inhibited luciferase production by 50% ($IC_{50}$) was calculated. Hygromycin, puromycin, and cycloheximide yielded an $IC_{50}$ of 0.008, 0.8, and $0.7{\mu}g/ml$, respectively. A filtrate of Streptomyces spp. isolates inhibited protein synthesis up to S-fold when added to the in vitro translation assay mixture.

Synthesis and Biological Evaluation of New Aminothiazolyl Cephalosporins with Elongated Side Chains

  • Kang, Han-Young;Koh, Hun-Yeong;Kim, Mi-Young;Kim, Hyun-Mi;Yoon, Du-Weon;Chang, Moon-Ho
    • Bulletin of the Korean Chemical Society
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    • v.12 no.6
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    • pp.666-673
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    • 1991
  • The elongation of the chain length at 2'position of aminothiazolylacetamido group of cephalosporin antibiotics was achieved. The derivatives with elongated side chain at 2'position were successfully synthesis and their in vitro activities were evaluated. It is indicated that there is an optimum range of the chain length in order to exhibit higher biological activities. This result could open a possibility for the development of the cephalosporin antibiotics with higher activities by further variation of the substituents at other positions. The cephalosporin derivatives with a simple (relatively non-functionalized) side chain as well as with a linear extended side chain were also prepared. Low activities of these derivatives led to a conclusion that either simple (non-functionalized) change or linear elongation of the side chain does not enhance the antibacterial activities.

Cytotoxicity of T-2 Toxin on Primary Cultures of Rat Hepatocytes

  • Kim, Hwan-Mook;Kim, Byung-Sam;Choe, Suck-Young;Yang, Kyu-Hwan
    • Toxicological Research
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    • v.4 no.1
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    • pp.37-45
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    • 1988
  • Primary cultures of adult rat hepatocytes were used to study in vitro cytotoxic effects of T-2 toxin on liver cells. When T-2 toxin was added to the culture, a significant depression of the hormonal induction of ${\alpha}$-aminoisobutyric acid (AIB) uptake and tyrosine aminotransferase (TAT) activity was observed. However, T-2 toxin did not affect the uptake of ouabain into hepatocytes. Protein synthesis was inhibited by T-2 toxin, but RNA synthesis was not severely affected. The inhibitory effects of T-2 toxin on protein synthesis was diminished rapidly with culture time and the hepatocytes culture maintained control level of protein synthesis within 24 hrs.

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A Facile and Efficient Synthesis of Dronedarone Hydrochloride

  • Li, Feng;Jin, Chunhua;Zou, Jianwei;Wu, Jun
    • Bulletin of the Korean Chemical Society
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    • v.35 no.7
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    • pp.1970-1972
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    • 2014
  • A facile and efficient synthesis of dronedarone hydrochloride starting from commercially available 4-nitrophenol is described. This approach features a tandem-type synthesis of 3-carbonylated benzofuran involving cyclization of 2-ethynylphenol followed by $CO_2$ fixation at the 3-position of the benzofuran ring mediated by potassium carbonate without the addition of any transition metal catalyst.

DETECTION OF DNA SINGLE-STRAND BREAKS AND UNSCHEDULED DNA SYNTHESIS INDUCED BY PROCARCINOGENS IN PRIMARY CULTURES OF RAT HEPATOCYTES

  • Kim, D.H.;Kim, Bok-Ryang;K. H. Yang
    • Toxicological Research
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    • v.2 no.1
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    • pp.1-7
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    • 1986
  • Procarcinogen induced DNA single-strand breaks and unschduled DNA synthesis were measured in primary rat hepatocytes culture. For DNA single-strand breaks assay, rat liver DNA was prelabeled by injection 3H-thymidine during the peak of DNA synthesis following partial hepatectomy. Hepatocytes were isolated from the rat 2 weeks after surgery by a collagenase perfusion techinique and maintained as monolayers in serum free medium on collagen-coated culture dishes. DNA sigle-strand breaks were measured by the alkaline elution techinique.

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