• Title/Summary/Keyword: Biological Synthesis

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X-ray Scattering Studies on the Structure of Porcine Pepsin in Various pH Solutions

  • Jin, Kyeong-Sik;Jin, Sang-Woo;Yoon, Jin-Hwan;Heo, Kyu-Young;Kim, Jae-Han;Kim, Hee-Soo;Ree, Moon-Hor
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.348-348
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    • 2006
  • In the present study, porcine pepsin in various pH conditions was investigated by small-angle X-ray scattering (SAXS) in order to detailed information on the structure and its variations with pH conditions. These analyses identified the structure of pepsin, and compared with that obtained by single crystallography. Moreover, this study found the structure' variations with changing pH conditions. All the results will be discussed with considering the conformational characteristics of pepsin in solution and further correlate to the biological and spectroscopic characteristics reported previously.

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Presence of Pituitary Specific Transcription Factor Pit-1 in the Rat Brain: Intracerebroventricular Administration of Antisense Pit-1 Oligodeoxynucleotide Decreases Brain Prolactin mRNA Level

  • Tae Woo Kim;Hyun-Ju Kim;Byung Ju Lee
    • Animal cells and systems
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    • v.3 no.3
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    • pp.311-317
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    • 1999
  • Prolactin (PRL) was reported to be locally synthesized in many brain areas including the hypothalamus, thalamus (TH) and hippocampus (HIP). In the pituitary lactotrophs, PRL synthesis is dependent upon a pituitary-specific transcription factor, Pit-1. In the present study, we attempted to identify Pit-1 or Pit-1-like protein in brain areas known as the synthetic sites of PRL. Reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis showed the same Pit-1 transcripts in brain areas such as the medial basal hypothalamus (MBH), preoptic area (POA), TH, and HIP with the Pit-1 transcripts in the anterior pituitary (AP). Electrophoretic mobility shift assay (EMSA) was run with nuclear protein extracts from brain tissues using a double strand oligomer probe containing a putative Pit-1 binding domain. Shifted bands were found in EMSA results with nuclear proteins from MBH, POA, TH and HIP. Specific binding of the Pit-1-like protein was further confirmed by competition with an unlabeled cold probe. Antisense Pit-1 oligodeoxynucleotide (Pit-1 ODN), which was designed to bind to the Pit-1 translation initiation site and block Pit-1 biosynthesis, was used to test Pit-1 dependent brain PRL transcription. Two nmol of Pit-1 ODN was introduced into the lateral ventricle of a 60-day old male rat brain. RNA blot hybridization and in situ hybridization indicated a decrease of PRL mRNA signals by the treatment of Pit-1 ODN. Taken together, the present study suggests that Pit-1 may play an important role in the transcriptional regulation of local PRL synthesis in the brain.

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Maximization of Poly-$\beta$-Hydroxybutyrate Accumulation by Potassium Limitation in Methylobacterium organophilum and Its Related Metabolic Analysis

  • Kim, Seon-Won;Kim, Pil;Kim, Jung-Hoe
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.140-146
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    • 1999
  • When methanol was the sole carbon source, Methylobacterium organophilum NCIB 11278, a facultative methylotroph, accumulated Poly-$\beta$-hydroxybutyrate (PHB) as 59% (w/w) of dry cell weight under potassium limitation, 37% under sulfate limitation, and 33% under nitrogen limitation. Based on a stoichiometric analysis of PHB synthesis from methanol, it was suspected that PHB synthesis is accompanied by the overproduction of energy, either 6-10 ATP and 1 $FADH_2$ or 6 ATP and 3 NADPH to balance the NADH requirement, per PHB monomer. This was confirmed by observation of increased intracellular ATP levels during PHB accumulation. The intracellular ATP with limited potassium, sulfate, and ammonium increased to 0.185, 0.452, and 0.390 $\mu$moles ATP/g Xr (residual cell mass) during PHB accumulation, respectively. The intracellular ATP level under potassium limitation was similar to that when there was no nutrient limitation and no PHB accumulation, 0.152- 0.186 $\mu$moles ATP/g Xr. We propose that the maximum PHB accumulation observed when potassium was limited is a result of the energy balance during PHB accumulation. Microorganisms have high energy requirements under potassium limitation. Enhanced PHB accumulation, in ammonium and sulfate limited conditions with the addition of 2,4-dinitrophenol, which dissipates surplus energy, proves this assumption. With the addition of 1 mM of 2,4-dinitrophenol, the PHB content increased from 32.4% to 58.5% of dry cell weight when nitrogen limited and from 15.1 % to 31.0% of dry cell weight when sulfate limited.

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Synthesis and Biological Activity Test of the Sex Pheromone of the Diamond Back Moth (배추좀나방의 성 페로몬의 합성과 생물활성시험)

  • Suk-Ku Kang;Chul-Hee Lee;Jung Han Kim;Jeong-Oon Lee
    • Journal of the Korean Chemical Society
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    • v.32 no.1
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    • pp.60-64
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    • 1988
  • Synthesis and biological activity test are described for the (Z)-11-hexadecen-1-ol, (Z)-11-hexadecen-1-yl acetate, and (Z)-11-hexadecen-l-al, the sex pheromone of the diamond back moth, Plutella xylostella L.. Lithium acetylide was alkylated with 10-bromodecan-1-ol THP ether to give 11-hexadecyn-l-ol THP ether. 11-Hexadecyn-l-ol THP ether was stereoselectively reduced over Pd/BaSO4to yield (Z)-11-hexadecen-l-ol THP ether, which was in turn deprotected to provide (Z)-11-hexadecen-l-ol. (Z)-11-Hexadecen-l-ol was acetylated and oxidized to afford (Z)-11-hexadecen-1-yl acetate and (Z)-11-hexadecen-l-al, respectively. Biological activity test of the synthetic compounds, (Z)-11-hexadecen-l-ol, (Z)-11-hexadecen-1-yl acetate, and (Z)-11-hexadecen-l-al in the ratio of 0.1 : 5 : 5 was tested in the field using polyethylene capsules as containers. The numbers of moth trapped with pheromone vials were counted.

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Cholera Toxin B Subunit-Porphyromonas gingivalis Fimbrial Antigen Fusion Protein Production in Transgenic Potato

  • Lee, Jin-Yong;Kim, Mi-Young;Jeong, Dong-Keun;Yang, Moon-Sik;Kim, Tae-Geum
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.268-274
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    • 2009
  • Porphyromonas gingivalis, the gram-negative anaerobic oral bacterium, initiates periodontal disease by binding to saliva-coated oral surface. The cholera toxin B subunit (CTB) genetically linked to FimA1 (1-200 aa) or FimA2 (201-337 aa) of the P. gingivalis fimbrial antigen were introduced into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation method. The integration of CTB-FimA1 or CTB-FimA2 fusion genes were confirmed in the chromosome of transformed leaves by genomic DNA PCR amplification method. Synthesis and assembly of the CTB-FimA fusion proteins into oligomeric structures with pentamer size was detected in transformed tuber extracts by immunoblot analysis. The binding activities of CTB-FimA fusion proteins to intestinal epithelial cell membrane receptors were confirmed by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). The ELISA showed that the expression levels of the CTB-FimA1 or CTB-FimA2 fusion proteins were 0.0019, 0.002% of the total soluble protein in transgenic tuber tissues, respectively The synthesis of CTB-FimA monomers and their assembly into biologically active oligomers in transformed potato tuber tissues demonstrates the feasibility of using edible plants for the production of enterocyte targeted fimbrial antigens that could elicit mucosal immune responses.

Biological Effects Of Flurbiprofen Loaded Chitosan To Gingival Fibroblast (Flurbiprofen 함유 키토산 제제가 치은 섬유아세포에 미치는 영향)

  • Chung, Chong-Pyoung;Park, Yoon-Jeong;Lee, Seung-Jin;Rhyu, In-Cheol;Choi, Sang-Mook
    • Journal of Periodontal and Implant Science
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    • v.26 no.1
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    • pp.317-333
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    • 1996
  • The main goal of periodontal regeneration is to be achieved by epithelial exclusion, periodontal ligament cell activation or alveolar bone regeneration. The purpose of this study was to investigate on the physico- chemical and biological characteristics of biodegradable chitosan beads. Chitosan beads were fabricated by ionic gelation with sodium tripolyphosphate and they had the size in 300um diameter. As therapeutic agent, flurbiprofen was incorporated into the beads by 10, 20% loading contents. The release of drugs from the chitosan beads was measured in vitro. Also, biological activity tests of flurbiprofen loaded chitosan beads including cytotoxicity test, ihhibition of $IL-1{\beta}$ production, suppression to $PGE_2$ production, collagenase inhibition test, the ability of total protein synthesis, and tissue response were evaluated. The amount of flurbiprofen released from chitosan was 33-50% during 7 days. Minimal cytotoxicity was observed in chitosan beads. Flurbiprofen released from chitosan beads significantly suppressed the $IL-1{\beta}$ production of monocyte, $PGE_2$ production and markedly inhibited collagenase activity. Meanwhile, flurbiprofen released from this system showed increased ability for protein synthesis. Throughout 4 -week implantation period, no significant inflammatory cell infiltrated around chitosan bead and also fibroblast like cell types at the beads - tissue interface were revealed with gradual degradation of implanted chitosan beads. From these results, it was suggested that flurbiprofen loaded chitosan beads can be effectively useful for biocompatible local delivery system in periodontal regeneration.

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Genetic Synthesis and Applications of Repetitive Protein Polymers (반복단위 단백질 고분자의 유전공학적 합성 및 응용)

  • Park, Mi-Sung;Choi, Cha-Yong;Won, Jong-In
    • KSBB Journal
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    • v.22 no.4
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    • pp.179-184
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    • 2007
  • This study introduces the characteristics and some applications of repetitive polypeptides, especially to the biomaterial, tissue engineering scaffolds, drug delivery system, and DNA separation systems. Since some fibrous proteins, which consist of repeating peptide monomers, have been reported that their physical properties are changed dramatically by means of temperature alteration or pH shifting. For that reason, fibrous protein-mimetic polypeptides, which are produced by the recombinant technology, can be applied to the diverse biological fields. Repetitive polypeptides can also be used in the bioseparation area such as DNA sequencing, because they make DNA separation possible in free-solution electrophoresis by conjugating DNA fragments to them. Moreover, artificial synthesis of repetitive polypeptides helps to demonstrate the correlations between mechanical properties and structures of natural protein polymer, which have been proven that repetitive domains are affected by the sequence of the repeating domains and the number of repeating subunits. Repetitive polypeptides can be biologically synthesized using some special cloning methods, which are represented here. Recursive directional ligation (RDL) and controlled cloning method (CCM) have been proposed as excellent cloning methods in that we can control the number of repetition in the multimerization of polypeptides and the components of repetitive polypeptides by either method.

Synthesis of Biodiesel Using Supercritical Fluid (초임계유체를 이용한 바이오디젤연료의 제조기술)

  • Lee, Youn-Woo;Song, Eun-Seok;Kim, Hwayong
    • Clean Technology
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    • v.11 no.4
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    • pp.171-179
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    • 2005
  • Biodiesel is synthesized by transesterification of vegetable or animal oils with alcohols. Since it has similar characteristic with diesel fuel, it can be used as a fuel by mixing with diesel fuel. Moreover, it is advantageous that biodiesel can reduce air pollution emitted from fuel combustion and is produced from sustainable energy, biomass. Recently, many researchers have investigated biodiesel synthesis using supercritical methanol since it is economical due to shorter reaction time and simple separation/purification process, compared with conventional alkali- or acid-catalyzed process. By the development of biodiesel production process from waste edible oil using supercritical methanol, it can be expected to utilize potential energy resources, reduce carbon dioxide emission, and improve environmental conditions.

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Identification and Characterization of a RecA-like Protein Induced by DNA Damaging Agents in Fluorescent Pseudomonas sp.

  • Kim, Ok-Bong;Lim, Chae-Kwang;Kim, Si-Wouk;Park, Jong-Kun;Yoon, Seong-Myeong;Lee, Jung-Sup
    • Animal cells and systems
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    • v.2 no.3
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    • pp.383-388
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    • 1998
  • A RecA-like protein (RecAps) was identified from fluorescent Pseudomonas sp. and the inducible nature of the protein was characterized in detail. It was shown by dose-response and time-course experiments using two DNA damaging agents, nalidixic acid and mitomycin-C, that the cellular level of RecAps protein was increased 3-8 fold compared to that of the control. The most effective doses of nalidixic acid and mitomycin-C for the protein induction were $30{\mu}g/ml$ and $0.3{\mu}g/ml$ at the treatment time point of 150 min, respectively. The enhanced level of RecAps protein was gradually decreased to the control level after 10 hr in normal medium. Interestingly, the cellular level of RecAps protein was increased by the same DNA damaging agents even when cell growth was completely inhibited by treatment with $170{\mu}g/ml$ of chloramphenicol, an inhibitor of protein synthesis, suggesting that new protein synthesis is not required for the induction of RecAps. All these results suggest that a typical S0S repair function driven by RecA-like protein is conserved in Pseudomonas sp. cells as in E, coli.

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Green synthesis of silver nanoparticles to the microbiological corrosion deterrence of oil and gas pipelines buried in the soil

  • Zhi Zhang;Jingguo Du;Tayebeh Mahmoudi
    • Advances in nano research
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    • v.15 no.4
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    • pp.355-366
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    • 2023
  • Biological corrosion, a crucial aspect of metal degradation, has received limited attention despite its significance. It involves the deterioration of metals due to corrosion processes influenced by living organisms, including bacteria. Soil represents a substantial threat to pipeline corrosion as it contains chemical and microbial factors that cause severe damage to water, oil, and gas transmission projects. To combat fouling and corrosion, corrosion inhibitors are commonly used; however, their production often involves expensive and hazardous chemicals. Consequently, researchers are exploring natural and eco-friendly alternatives, specifically nano-sized products, as potent corrosion inhibitors. This study aims to environmentally synthesize silver nanoparticles using an extract from Lagoecia cuminoides L and evaluate their effectiveness in preventing biological corrosion of buried pipes in soil. The optimal experimental conditions were determined as follows: a volume of 4 ml for the extract, a volume of 4 ml for silver nitrate (AgNO3), pH 9, a duration of 60 minutes, and a temperature of 60 degrees Celsius. Analysis using transmission electron microscopy confirmed the formation of nanoparticles with an average size of approximately 28 nm, while X-ray diffraction patterns exhibited suitable peak intensities. By employing the Scherer equation, the average particle size was estimated to be around 30 nm. Furthermore, antibacterial studies revealed the potent antibacterial activity of the synthesized silver nanoparticles against both aerobic and anaerobic bacteria. This property effectively mitigates the biological corrosion caused by bacteria in steel pipes buried in soil.