• Title/Summary/Keyword: $\mu$ synthesis

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Characterization of a Psychrophilic Metagenome Esterase EM2L8 and Production of a Chiral Intermediate for Hyperlipemia Drug (메타게놈유래의 저온성 에스터라제 EM2L8의 효소적 특성과 이를 활용한 고지혈증 치료제 키랄소재의 생산)

  • Jung, Ji-Hye;Choi, Yun-Hee;Lee, Jung-Hyun;Kim, Hyung-Kwoun
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.118-124
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    • 2009
  • Esterase EM2L8 gene isolated from deep sea sediment was expressed in Escherichia coli BL21 (DE3) and the esterase activity of the cell-free extract was assayed using p-nitrophenyl butyrate-spectrophotometric method. Its optimum temperature was $40-45^{\circ}C$ and 45% activity of the maximum activity was retained at $15^{\circ}C$. The activation energy at $15-45^{\circ}C$ was calculated to be 4.9 kcal/mol showing that esterase EM2L8 was a typical cold-adapted enzyme. Enzyme activity was maintained for 6 h and 4 weeks at $30^{\circ}C$ and $4^{\circ}C$, respectively. When each ethanol, methanol, and acetone was added to the reaction mixture to 15% concentration, enzyme activity was maintained. In the case of DMSO, enzyme activity was kept up to 40% concentration. (S)-4-Chloro-3-hydroxy butyric acid is a chiral intermediate for the synthesis of Atorvastatin, a hyperlipemia drug. When esterase EM2L8 (40 U) was added to buffer solution (1.2 mL, pH 9.0) containing ethyl-(R,S)-4-chloro-3-hydroxybutyrate (38 mM), it was hydrolyzed into 4-chloro-3-hydroxy butyric acid with a rate of $6.8\;{\mu}mole/h$. The enzyme hydrolyzed (S)-substrate more rapidly than (R)-substrate. When conversion yield was 80%, e.e.s value was 40%. When DMSO was added, hydrolysis rate increased to $10.4\;{\mu}mole/h$. The plots of conversion yield vs e.e.s in the presence or absence of DMSO were almost same, implying that the reaction enantioselectivity was not changed by the addition of DMSO. Taken together, esterase EM2L8 had high activity and stability at low temperatures as well as in various organic solvents/aqueous solutions. These properties suggested that it could be used as a biocatalyst in the synthesis of useful pharmaceuticals.

Disturbance of $\alpha$-Amylase Secretion from Bacillus amyloliquefaciens Cells by the Treatment of Puromycin and Magnesium (Bacillus amyloliquefaciens에서 Puromycin 과 Magnesium에 의한 $\alpha$-Amylase 의 분비저해)

  • 안순자;김순옥;이동희;송방호
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.412-420
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    • 1989
  • To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.

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Silica and Iron Oxide Recovery and Mineral Carbonation from Serpentine Minerals Using Acid Dissolution and pH Swing Processes (산 처리와 pH 조절을 이용한 사문석군 광물로부터 규소와 철산화물 회수 및 광물 탄산화 연구)

  • Baek, Jiyeon;Jo, Yeonu;Lee, Jeongheon;Kwon, Nayoon;Kim, Yeram;Choi, Suk;Kim, Sunghee;Roh, Yul
    • Economic and Environmental Geology
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    • v.49 no.1
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    • pp.13-22
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    • 2016
  • The objectives of this study were to recover silica and iron oxides and $CO_2$ sequestration using serpentine via various acid dissolution and pH swing processes. Serpentine collected from Guhang-myeon in S. Korea were mainly composed of antigorite and magnetite consisting of $SiO_2$ (45.3 wt.%), MgO (41.3 wt.%), $Fe_2O_3$ (12.2 wt.%). Serpentine pulverized ($${\leq_-}75{\mu}m$$) and then dissolved in 3 different acids, HCl, $H_2SO_4$, $HNO_3$. Residues treated with acidic solution were recovered from the solution (step 1). And then the residual solution containing dissolved serpentine was titrated using $NH_4OH$. And pH of the solution increased up to pH=8.6 to obtain reddish precipitates (step 2). After recovery of the precipitates, the residual solution reacted with $CO_2$ and then pH increased up to pH=9.5 to precipitate white materials (step 3). The mineralogical characteristics of the original sample and harvested precipitates were examined by XRD, and TEM-EDS analyses. ICP-AES analysis was also used to investigate solution chemistry. The dissolved ions were Mg, Si, and Fe. The antigorite became noncrystralline silica after acid treatment (step 1). The precipitate at pH=8.6 was mainly amorphous iron oxide, of which size ranged from 2 to 10 nm and mainly consisting of Fe, O, and Si (step 2). At pH=9.5, nesquehonite [$Mg(HCO_3)(OH){\cdot}2(H_2O)$] and lasfordite [$MgCO_3{\cdot}H_2O$] were formed after reaction with $CO_2$ (step 3). The size of carbonated minerals was ranged from 1 to $6{\mu}m$. These results indicated that the acid treatment of serpentine and pH swing processes for the serpentine can be used for synthesis of other materials such as silica, iron oxides and magnesium carbonate. Also, This process may be useful for the precursor synthesis and $CO_2$ sequestration via mineral carbonation.

Studies on the Varietal Features for the Silk Yielding Ability, Bombyx mori L. (가잠의 견사물질 생성능력에 있어서 품종간의 특이성에 관한 연구)

  • 이용우
    • Journal of Sericultural and Entomological Science
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    • v.22 no.1
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    • pp.26-45
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    • 1980
  • Silk protein is synthesized in the silkgland of silkworm, Bombyx mori L. It is evident that silk productivity is one of the high heritable characters from the genetical aspects. It is also changed with the environmental circumstances. With this regard, this paper dealt with the varietal patterns of silkgland development and the factors concerning to the silk productivity of silkgland of silkworm by the synthesis of nucleic acids, profiles of amino acids and histological basis, using the eight parent silkworm varieties and their F$_1$ hybrids. 1. The weight of silkgland per larva increased proportionally in the F$_1$ hybrids which were crossed between longer silk yielding varieties. The silk content to the weight of the silkgland was higher in the longer silk yielding varieties than that in the shorter silk yielding varieties. 2. It was observed that the morphological changes of nuclei took place in the posterior silkgland cells with the larval development of the 5th instar. In varietal aspect, Jam 107 and Jam 108, longer silk yielding varieties, showed more branches in nuclei than the $N_2$ and $C_{60}$ which were shorter yielding ones. 3. It was observed that there was a high correlation between RNA content per unit weight of silkgland on the 6th day stage of 5th instar and silk productivity both in the parents and their F$_1$ hybrids. 4. RNA and DNA synthesis brought about thirty percent increase in the posterior silkgland of the longer silk yielding varieties during the 2nd day to the 4th day stages of the 5th instar, when compared with those in the posterior silkgland of the shorter silk yielding varieties. 5. RNA/DNA ratio in the posterior silkgland on the 2nd day and 4th day stages of the 5th instar was more increased in the longer silk yielding varieties than the shorter silk yielding varieties. 6. It was shown that DNA content for the longer silk yielding varieties came to be 374$\mu\textrm{g}$ per larva in the posterior division of silkgland on the 4th day stage of 5th instar, whereas it was 199$\mu\textrm{g}$ per larva for the shorter silk yielding varieties. 7 There was 34.8% Alanine, 22.8% Glycine, 9.1% Serine and 7.3% Tyrosine in the posterior division of silkgland as major amino acids. It is noticed that there was a little differences between the amino acids composition of posterior silkgland and silk fibroin. 8. There was some differences in the amino acids composition of posterior silkgland between pure lines and their hybrids. Glycine, Serine and acidic amino acids, essential to silk formation, seemed to be increased in the F$_1$ hybrids, whereas other amino acids such as Valine, Iso-leucine, Leucine, Lysine. Phenylalanine, Histidine and Arginine were reduced. 9. The content of Glycine, Alanine and Serine in the posterior division of silkgland was elevated in the longer silk yielding varieties than the others. Consequently. these three amino acids in the posterior silkgland seemed to be related to the silk yielding ability in the longer silk yielding varieties.s.

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Facile [11C]PIB Synthesis Using an On-cartridge Methylation and Purification Showed Higher Specific Activity than Conventional Method Using Loop and High Performance Liquid Chromatography Purification (Loop와 HPLC Purification 방법보다 더 높은 비방사능을 보여주는 카트리지 Methylation과 Purification을 이용한 손쉬운 [ 11C]PIB 합성)

  • Lee, Yong-Seok;Cho, Yong-Hyun;Lee, Hong-Jae;Lee, Yun-Sang;Jeong, Jae Min
    • The Korean Journal of Nuclear Medicine Technology
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    • v.22 no.2
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    • pp.67-73
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    • 2018
  • $[^{11}C]PIB$ synthesis has been performed by a loop-methylation and HPLC purification in our lab. However, this method is time-consuming and requires complicated systems. Thus, we developed an on-cartridge method which simplified the synthetic procedure and reduced time greatly by removing HPLC purification step. We compared 6 different cartridges and evaluated the $[^{11}C]PIB$ production yields and specific activities. $[^{11}C]MeOTf$ was synthesized by using TRACERlab FXC Pro and was transferred into the cartridge by blowing with helium gas for 3 min. To remove byproducts and impurities, cartridges were washed out by 20 mL of 30% EtOH in 0.5 M $NaH_2PO_4$ solution (pH 5.1) and 10 mL of distilled water. And then, $[^{11}C]PIB$ was eluted by 5 mL of 30% EtOH in 0.5 M $NaH_2PO_4$ into the collecting vial containing 10 mL saline. Among the 6 cartridges, only tC18 environmental cartridge could remove impurities and byproducts from $[^{11}C]PIB$ completely and showed higher specific activity than traditional HPLC purification method. This method took only 8 ~ 9 min from methylation to formulation. For the tC18 environmental cartridge and conventional HPLC loop methods, the radiochemical yields were $12.3{\pm}2.2%$ and $13.9{\pm}4.4%$, respectively, and the molar activities were $420.6{\pm}20.4GBq/{\mu}mol$ (n=3) and $78.7{\pm}39.7GBq/{\mu}mol$ (n=41), respectively. We successfully developed a facile on-cartridge methylation method for $[^{11}C]PIB$ synthesis which enabled the procedure more simple and rapid, and showed higher molar radio-activity than HPLC purification method.

Synthesis and Antifungal Activity of 6,7-Bis-[S-(Aryl)thio]-5,8-Quinolinediones

  • Ryu, Chung-Kyu;Sun, Yang-Jung;Shim, Ju-Yeon;You, Hea-Jung;Choi, Ko-Un;Lee, Hee-Soon
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.795-800
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    • 2002
  • 6,7-Bis-[S-(aryl)thio]-5,8-quinolinediones 4 and 5 were synthesized by the substitution of 6,7-dichloro-5,8-quinolinediones with appropriate arylthiols. Their antifungal activity were tested in vitro for their growth inhibitory activities against pathogenic fungi in comparison with flucytosine. The antifungal activities were significantly improved by S-(aryl)thio moieties of the compounds 4 and 5. The all tested compounds 4 and 5 showed generally good activities against C. albicans and A. niger ranging from 0.8 to 25 $\mu\textrm{g}$/ml. Among them, compounds 4d-4h and 5a-5c exhibited also good activities against C. krusei and C. tropicalis. The activities of compounds 4j and 4I were comparable to those of flucytosine against all tested fungi.

Comparison of Optical Properties of Ga-doped and Ag-doped ZnO Nanowire Measured at Low Temperature

  • Lee, Sang Yeol
    • Transactions on Electrical and Electronic Materials
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    • v.15 no.5
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    • pp.262-264
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    • 2014
  • Pristine ZnO, 3 wt.% Ga-doped (3GZO) and 3 wt.% Ag-doped (3SZO) ZnO nanowires (NWs) were grown using the hot-walled pulse laser deposition (HW-PLD) technique. The doping of Ga and Ag in ZnO NWs was observed by analyzing the optical and chemical properties. We optimized the synthesis conditions, including processing temperature, time, gas flow, and distance between target and substrate for the growth of pristine and doped ZnO NWs. The diameter and length of pristine and doped ZnO NWs were controlled under 200 nm and several ${\mu}m$, respectively. Low temperature photoluminescence (PL) was performed to observe the optical property of doped NWs. We clearly observed the shift of the near band edge (NBE) emission by using low temperature PL. In the case of 3GZO and 3SZO NWs, the center photon energy of the NBE emissions shifted to low energy direction using the Burstein Moss effect. A strong donor-bound exciton peak was found in 3 GZO NWs, while an acceptor-bound exciton peak was found in 3SZO NWs. X-ray photoelectron spectroscopy (XPS) also indicated that the shift of binding energy was mainly attributed to the interaction between the metal ion and ZnO NWs.

A Study on the Antioxidative and Depigmentation Activities of the Ethanol Extract of Saururus Herba (삼백초(三白草) 에탄올추출물의 항산화능 및 피부 미백작용 연구)

  • Park, Dae-Jung;Lee, Jang-Cheon
    • The Korea Journal of Herbology
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    • v.23 no.2
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    • pp.193-202
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    • 2008
  • Objectives : The purpose of this study is to investigate the etahnol extract of Saururus Herba and their dermal boiactivity properties related to cosmeceuticals such as depigmentation and radical scavenging effect. Methods : Using B16/F10 Melanin cells, we measured depigmentation effect of the etahnol extract of Saururus Herba. Antioxidant activities of the etahnol extract of Saururus Herba were determined by DPPH radical scavenging activity. Results : The etahnol extract of Saururus Herba showed considerable radical scavenging activity($SC_{50}$ : 25 ${\mu}g$/ml). Antioxidant activity is in a dose dependant manner. The etahnol extract of Saururus Herba down-regulated melanin synthesis effectively. B16 cells incubated with the etahnol extract of Saururus Herba showed reduces melanin formation and tyrosinase activity. The etahnol extract of Saururus Herba alone markedly suppressed melanin content and tyrosinase activity. Conclusions : These results suggest that the ethanol extract of Saururus Herba is a potent depigmentaion agent and it may be a candidate for antioxidant agent.

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Facile Synthesis of Highly Dispersed Ultra-fine ZrC Powders by Carbothermal Reduction Method Using Nanosized ZrO2 and Nanosized Graphite Powder Mixtures (나노크기의 ZrO2와 Graphite 분말 혼합체의 열탄소환원법에 의한 고분산 초미립 ZrC 분말의 합성)

  • Lee, Wha-Jun;Ryu, Sung-Soo
    • Journal of Powder Materials
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    • v.20 no.2
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    • pp.100-106
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    • 2013
  • Ultra-fine zirconium carbide (ZrC) powder with nano-sized primary particles was synthesized by the carbothermal reduction method by using nano-sized $ZrO_2$ and nano-sized graphite powders mixture. The synthesized ZrC powder was well dispersed after simple milling process. After heat-treatment at $1500^{\circ}C$ for 2 h under vacuum, ultra-fine ZrC powder agglomerates (average size, $4.2{\mu}m$) were facilely obtained with rounded particle shape and particle size of ~200 nm. Ultra-fine ZrC powder with an average particle size of 316 nm was obtained after ball milling process in a planetary mill for 30 minutes from the agglomerated ZrC powder.

Synthesis of 1,6-Disubstituted 4,5,6,7-Tetrahydropyrazolo[3,4-c]pyridin-7-one Derivatives and Evaluation of Their Anticancer Activity

  • Devegowda, Vani Nelamane;Seo, Seon-Hee;Pae, Ae Nim;Nam, Ghil-Soo;Choi, Kyung-Il
    • Bulletin of the Korean Chemical Society
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    • v.33 no.2
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    • pp.647-650
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    • 2012
  • Promising anticancer compounds of the type 1,6-disubstituted 4,5,6,7-tetrahydropyrazolo[3,4-c]pyridin-7-ones were identified. The target compounds were readily synthesized in a large scale via a sequence of reactions starting from the commercially available primary amines. Their in vitro anti-proliferative activity has been evaluated on prostate (DU-145), colon (HT-29 and HCT-116) and melanoma (A375P) human cancer cell lines. The relationships between the structure and the anticancer activity, covering all tested cancer cell lines, revealed that the compound 5c with 2,4-dimethylphenyl substituent at $R^2$ was the most potent with the $IC_{50}$ values in the range as low as 0.16 to $0.40{\mu}M$.