• Title/Summary/Keyword: Biological synthesis

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Panax ginseng total protein promotes proliferation and secretion of collagen in NIH/3T3 cells by activating extracellular signal-related kinase pathway

  • Chen, Xuenan;Wang, Manying;Xu, Xiaohao;Liu, Jianzeng;Mei, Bing;Fu, Pingping;Zhao, Daqing;Sun, Liwei
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.411-418
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    • 2017
  • Background: Recently, protein from ginseng was studied and used for the treatment of several kinds of diseases. However, the effect of ginseng total protein (GTP) on proliferation and wound healing in fibroblast cells remains unclear. Methods: In this study, cell viability was analyzed using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Cell cycle distribution was analyzed by flow cytometer. The levels of transforming growth factor ${\beta}1$, vascular endothelial growth factor, and collagens were analyzed by enzyme-linked immunosorbent assay and immunofluorescence staining. The expressions of cyclin A, phosphorylation of extracellular signal-related kinase (p-ERK1/2), and ERK1/2 were analyzed by Western blotting. Results: Our results showed that GTP promoted cell proliferation and increased the percentage of cells in S phase through the upregulation of cyclin A in NIH/3T3 cells. We also found that GTP induced the secretion of type I collagen, and promoted the expression of other factors that regulate the synthesis of collagen such as transforming growth factor ${\beta}1$ and vascular endothelial growth factor. In addition, the phosphorylation of ERK1/2 at Thr202/Tyr204 was also increased by GTP. Conclusion: Our studies suggest that GTP promoted proliferation and secretion of collagen in NIH/3T3 cells by activating the ERK signal pathway, which shed light on a potential function of GTP in promoting wound healing.

Synthesis of L-threo-3,4-Dihydroxyphenylserine(L-threo-DOPS) with Thermostabilized Low-Specific L-Threonine Aldolase from Streptomyces coelicolor A3(2)

  • Baik, Sang-Ho;Yoshioka, Hideki;Yukawa, Hideaki;Harayama, Shigeaki
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.721-727
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    • 2007
  • Stability-enhanced mutants, H44, 11-94, 5A2-84, and F8, of L-threonine aldolase(L-TA) from Streptomyces coelicolor A3(2)(SCO1085) were isolated by an error-prone PCR followed by a high-throughput screening. Each of these mutant, had a single amino acid substitution: H177Y in the H44 mutant, A169T in the 11-94 mutant, D104N in the 5A2-84 mutant and F18I in the F8 mutant. The residual L-TA activity of the wild-type L-TA after a heat treatment for 20 min at $60^{\circ}C$ was only 10.6%. However, those in the stability-enhanced mutants were 85.7% for the H44 mutant, 58.6% for the F8 mutant, 62.1% for the 5A2-84 mutant, and 67.6% for the 11-94 mutant. Although the half-life of the wild-type L-TA at $63^{\circ}C$ was 1.3 min, those of the mutant L-TAs were longer: 14.6 min for the H44 mutant, 3.7 min for the 11-94 mutant, 5.8 min for the 5A2-84 mutant, and 5.0 min for the F8 mutant. The specific activity did not change in most of the mutants, but it was decreased by 45% in the case of mutant F8. When the aldol condensation of glycine and 3,4-dihydroxybenzaldehyde was studied by using whole cells of Escherichia coli containing the wild-type L-TA gene, L-threo-3,4-dihydroxyphenylserine(L-threo-DOPS) was successfully synthesized with a yield of 2.0 mg/ml after 20 repeated batch reactions for 100 h. However, the L-threo-DOPS synthesizing activity of the enzyme decreased with increased cycles of the batch reactions. Compared with the wild-type L-TA, H44 L-TA kept its L-threo-DOPS synthesizing activity almost constant during the 20 repeated batch reactions for 100 h, yielding 4.0 mg/ml of L-threo-DOPS. This result showed that H44 L-TA is more effective than the wild-type L-TA for the mass production of L-threo-DOPS.

EFFECT OF QUINONES ON $NADPH_2$ OXIDATION AND PHOTOSYNTHETIC $CO_2$ ASSIMILATION IN CHLORELLA PYRENOIDOSA (Chlorella Pyrenoidosa에 있어서 $NADPH_2$의 산화(酸化)와 광합성(光合性)에 의(依)한 $CO_2$ 동화작용(同化作用)에 대(對)한 Quinone의 영향(影響))

  • Cho, Duck-Hiyon
    • Applied Biological Chemistry
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    • v.11
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    • pp.29-33
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    • 1969
  • 1. The oxidizability of $NADPH_2$ by quinones in the presence of $NADPH_2$-diaphorase was tested under aerobic conditions. Also the $^{14}CO_2$-fixation rates were compared when Chlorella suspensions were pretreated with $3{\cdot}10^{-5}M$ cocentration of variou quinones for 10 minutes prior and during the $^{14}CO_2$-fixation period. 2. A close correlation seems to exist between the rate of $NADPH_2$ oxidation by quinones and the $^{14}CO_2$-fixation rate. The effect of quinones on $NADPH_2$ oxidation and $^{14}CO_2$-fixation were in the order of Dichlone>06-K>NQ>BQ. 3. It is postulated that the phytotoxicity of quinones on Chlorella is due to the deprival of $NADPH_2$ consequently inhibiting $^{14}CO_2$-fixation, thus causing death of the cells. 4. The effect of quinones on amino acids biosyn-thesis in Chlorella was one of depressed rates, which was especially noted in the case of dichlone. This would be expected from a consideration of $NADPH_2$ deprival and inhibition of $^{14}CO_2$-fixation. Sucrose synthesis was either not affected or rather stimulated, the reasons of which are not clear at the present time.

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Effects of Some Plant Growth Regulators on Protein Biosynthesis of Carrot Cells (당근 세포의 단백질 생합성에 대한 몇가지 식물생장조절제의 영향)

  • Yoo, Ki-Jung;Park, Chang-Kyu;Song, Tae-Chul
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.187-195
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    • 1985
  • Electrophoretic studies of protein extracts from carrot calluses suspension-cultured on the media containing kinetin, BA, IAA, NAA or $GA_3$ at the levels of $10^{-6},\;10^{-5},\;10^{-4}M$, respectively, were performed to identify polypeptides and proteins regulated by auxin, cytokinin or GA. Fifteen bands of polypeptide(s) were observed in the callus cultured in the control medium devoid of growth regulators, and their molecular weights were $18._4,\;20._2,\;20._0,\;34._9,\;35._7,\;37._4,\;40._3,\;42._2,\;44._1,\;44._4,\;49._3,\;55._0,\;56._6,\;58._1,\;and\;59._9\;KD$, respectively. The synthesis of polypeptide appeared to be promoted in two bands by kinetin, in six bands by BA, in one band by IAA, in two bands by NAA, and in four bands by $GA_3$, while inhibited in five bands by kinetin, in three bands by BA, in four bands by IAA, in three bands by NAA and in three bands by $GA_3$. The polypeptides of $40._3\;KD\;42._2\;KD$ seemed to be regulated by cytokinins, and those of $44._1\;KD,37._4\;KD,\;and\;56._6\;KD$ by auxins. The proteins of three bands with relative mobilities of 0.56, 0.84, and 0.92, respectively, increased in the calluses cultured on the media containing kinetin, IAA, $GA_3$, NAA or BA, compared to the control, but it was difficult to identify the proteins specific for each growth regulator.

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Methanol Extract of Alpinia katsumadai Hayata has Anti-viral Activity by Inhibition of α-glucosidase (초두구 추출물의 α-glucosidase 활성 억제에 의한 항바이러스 활성)

  • Lee, Doseung;Boo, Kyung Hwan;Kim, Young Cheon;Lee, Jin-Man;Kim, Seong Cheol;Lee, Wang Shik;Riu, Key Zung;Lee, Dong-Sun
    • Journal of Applied Biological Chemistry
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    • v.57 no.2
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    • pp.179-182
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    • 2014
  • In the present study, we investigated the effects of methanol extracts from Alpinia katsumadai Hayata against antiviral potential underlying mechanism in glucosidase inhibition. Syncytium formation in Newcastle disease virus (NDV)-infected baby hamster kidney (BHK) cell originates from the trafficking of viral glycoprotein into cell-surface. Methanol extracts inhibited not only syncytium formation, but also trafficking of glycoprotein, hemagglutinin-neuraminidase (HN), onto cell-surface. A. katsumadai extracts showed the inhibitory activities ($IC_{50}$ $25{\mu}g/mL$) against ${\alpha}$-glucosidase. These results suggested that blue chanterelle extracts inhibited the cell-surface expression of NDV-HN glycoprotein without significantly affecting HN glycoprotein synthesis in NDV-infected BHK cells.

Synthesis and Biological Activities of Myomodulin A and Its Analogs (Myomodulin A 및 유도체들의 합성 및 생리활성)

  • Park, Nam-Gyu
    • Journal of Life Science
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    • v.22 no.3
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    • pp.387-397
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    • 2012
  • In this study, we focused on myomoduline A (MMA) released from the central nervous system of Aplysia kurodai. The primary structure of MMA is Pro-Met-Ser-Met-Leu-Arg-Leu-$NH_2$. This peptide is the same as that of the myomodulin family peptide found in other mollusks. The purified MMA showed a modulating activity of phasic contraction on the anterior byssus retractor muscle (ABRM) of Mytilus edulis. In order to study the relationship between the structure and biological activity of MMA, we synthesized MMA, Des[$Pro^1$]-MMA, Des[$Pro^1,Met^2$]-MMA, Des[$Pro^1,Met^2,Ser^3$]-MMA, and MME. The amino acid sequences of Des[$Pro^1$]-MMA, Des[$Pro^1,Met^2$]-MMA, and Des[$Pro^1,Met^2,Ser^3$]-MMA were Met-Ser-Met-Leu-Arg-Leu-$NH_2$, Ser-Met-Leu-Arg-Leu-$NH_2$, and Met-Leu-Arg-Leu-$NH_2$, respectively. MMA and synthetic peptides were tested on ABRM in M. edulis as well as muscle preparations in Achatina fulica. At $1{\times}10^{-8}$ M or lower, MMA showed a potentiating effect on phasic contraction of the ABRM, but this peptide had an inhibitory effect at $1{\times}10^{-6}$ M or higher. Both MMA and its analogs stimulated a contractile response on the crop and a relaxed catch-relaxing response on the penial retractor muscle of A. fulica. These results suggest that Met-Leu-Arg-Leu-$NH_2$ in MMA is the minimum structure required for the regulation of the contraction of ABRM, as well as the reproductive and digestive activities of mollusks.

Synthesis of ITiO(Indium Titanium Oxide) particle by sol-gel and investigation on light transmittance of deposited ITiO thin film (졸-겔법에 의한 ITiO(Indium Titanium Oxide) 입자의 합성과 ITiO 박막의 광투과도 조사)

  • Go, Eun Ju;Kim, Sang Hern
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.4
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    • pp.705-716
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    • 2017
  • In this study, Indium-Titanium hydroxide particle with 0.5, 1.0, 1.5 wt% of $TiO_2$ were synthesized by sol process and adding the base, ITiO(Indium Titanium Oxide) particles were obtained by gelling at $200^{\circ}C$ and $500^{\circ}C$. The ITiO particle's size with gel process at $200^{\circ}C$ was smaller than ITiO particle's size with gel process $500^{\circ}C$. The ITiO particle with gel process at $200^{\circ}C$ was used to fabricate dense ITiO target. ITiO targets with 0.5, 1.0, 1.5 wt% of $TiO_2$ were fabricated and used to obtain ITiO thin films onto glass by sputtering. Among those sputtered ITiOs' thin films, ITiO thin film with 0.4 % of $O_2$ and 0.5 wt% of $TiO_2$ showed the lowest specific resistance, highest charge mobility and lowest carrier concentration. It was found the light transmittance of the ITiO film were increased highly compared to light transmittance of ITO (Indium Tin Oxide) thin film over Infrared wavelength ranges.

Effect of Cu-resistant Pseudomonas on growth and expression of stress-related genes of tomato plant under Cu stress (구리-오염 토양에서 토마토 식물의 생장과 스트레스-관련 유전자 발현에 미치는 구리-내성 Pseudomonas의 영향)

  • Kim, Min-Ju;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.257-264
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    • 2017
  • Pseudomonas veronii MS1 and P. migulae MS2 have several mechanisms of copper resistance and plant growth promoting capability, and also can alleviate abiotic stress in plant by hydrolysis of a precursor of stress ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC) by ACC deaminase. In 4-week pot test for tomato growth in soil contained 700 mg/kg Cu, inoculation of MS1 and MS2 significantly increased root and shoot lengths, wet weight and dry weight of tomato plants compared to those of uninoculated control. The inoculated tomato plants contained less amounts of proline that can protect plants from abiotic stress, and malondialdehyde, an oxidative stress marker than those of control. ACC synthase genes, ACS4 and ACS6, and ACC oxidase genes, ACO1 and ACO4, both involved in ethylene synthesis, were strongly expressed in Cu stressed tomato, whereas significantly reduced in tomato inoculated with MS1 and MS2. Also, a gene encoding a metal binding protein metallothionein, MT2 showed similar expression pattern with above genes. All these results indicated that these rhizobacteria could confer Cu resistance to tomato plant under Cu stress and allowed a lower level of Cu stress and growth promotion.

Synthesis of ceria by combination of spray pyrolysis, postheat, and ball-milling and its characterization (분무열분해, 후소성 및 볼밀링을 조합한 방법을 이용한 세리아의 합성 및 특성연구)

  • Kim, Hyun-Ik;Kim, Sang Pil;Song, Jae-Kyung;Kim, Sang Hern
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.4
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    • pp.1057-1072
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    • 2018
  • In this study, micro-sized $CeO_2$ particles were synthesized by spray pyrolysis, and EG(ethylene glycol) and CA(citric acid) as organic additives were added to obtain hollow and porous particle during spray pyrolysis, and characteristics of obtained ceria were investigated according to the amount of added organic additives. Spray pyrolysis, postheat and ball-milling were combined to give 6 paths. $CeO_2$ nano-sized particle was obtained by the path which has sequence of Spray Pyrolysis with 0.5 M of EG and CA${\rightarrow}$Post-heat${\rightarrow}$Ball-milling${\rightarrow}$Post-heat among 6 paths. The average particle size(24 nm with standard deviation of 3.8 nm) of $CeO_2$ nano-sized particle by TEM analysis is close to the primary particle size(20 nm) which was calculated by Debye-Scherrer equation. To investigate the morphological characteristics and structure of the synthesized nanoparticle powders, SEM(Scanning Electron Microscopy), XRD(X-Ray Diffractometer) and TEM(Transmission Electron Microscopy) were used.

Synthesis and biological evaluation of diagnostic reagent for prostate cancer using copper-64 radioisotope

  • Ahn, Heesu;Kim, Mi Hyun;Han, Sang Jin;Woo, Sang Keun;Kim, Jung Young;Lee, Kyu Chul;Lim, Il Han;Lee, Yong Jin
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.4 no.2
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    • pp.65-72
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    • 2018
  • Prostate specific membrane antigen (PSMA) is a cell surface membrane protein, which is overexpressed in most prostate cancer. Recently, PET imaging with $[^{68}Ga]$PSMA-HBED-CC has been widely used for the diagnosis of recurrent prostate cancer and the studies on the diagnostic potential of $^{64}Cu$-labeled PSMA ligands reported actively. In this study, we monitored with biological evaluation in vivo and PET imaging of $^{64}Cu$-labeled PSMA ligand ($[^{64}Cu]$PSMA-617). The radiolabelling efficiency and stability of $[^{64}Cu]$PSMA-617 were confirmed by radio-thin layer chromatography. The radiolabeling efficiency of $[^{64}Cu]$PSMA-617 showed over 95%, and stabilities of intact remained over 98% in both human and mouse serum for 48 h. In normal male mice, in vivo uptake of $[^{64}Cu]$PSMA-617 in several organs was measured at 2, 4, 6, 24, 48 h after injection. Rapid blood clearance was observed for $[^{64}Cu]$PSMA-617. The high uptake was observed in the lung, liver, intestines and kidneys at 2 h postinjection, but was low in the other organs (1-2 %ID/g) at 4 h. The dynamic PET/CT images of 22RV1 tumor-bearing nude mice were acquired during 60 min and additionally acquired 24 h and 48 h after injection. In dynamic PET images, $[^{64}Cu]$PSMA-617 uptake ratio in tumors versus muscle was increased as time elaplsed until 60 minutes and remained in tumors at 48 h. In these results, the PET/CT imaging using $[^{64}Cu]$PSMA-617 in prostate cancer is expected to be useful for the diagnosis and treatment of prostate cancer patients.