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Purification and Characterization of the Laccase Involved in Dye Decolorization by the White-Rot Fungus Marasmius scorodonius

  • Jeon, Sung-Jong;Lim, Su-Jin
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1120-1127
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    • 2017
  • Marasmius scorodonius secretes an extracellular laccase in potato dextrose broth, and this enzyme was purified up to 206-fold using $(NH_4)_2SO_4$ precipitation and a Hi-trap Q Sepharose column. The molecular mass of the purified laccase was estimated to be ~67 kDa by SDS-PAGE. The UV/vis spectrum of the enzyme was nontypical for laccases, and metal content analysis revealed that the enzyme contains 1 mole of Fe and Zn and 2 moles of Cu per mole of protein. The optimal pH for the enzymatic activity was 3.4, 4.0, and 4.6 with 2,2'-azino-bis(3-ethylbenzothazoline-6-sulfonate) (ABTS), guaiacol, and 2,6-dimethoxy phenol as the substrate, respectively. The optimal temperature of the enzyme was $75^{\circ}C$ with ABTS as the substrate. The enzyme was stable in the presence of some metal ions such as $Ca^{2+}$, $Cu^{2+}$, $Ni^{2+}$, $Mg^{2+}$, $Mn^{2+}$, $Ba^{2+}$, $Co^{2+}$, and $Zn^{2+}$ at a low concentration (1 mM), whereas $Fe^{2+}$ completely inhibited the enzymatic activity. The enzymatic reaction was strongly inhibited by metal chelators and thiol compounds except for EDTA. This enzyme directly decolorized Congo red, Malachite green, Crystal violet, and Methylene green dyes at various decolorization rates of 63-90%. In the presence of 1-hydroxybenzotriazole as a redox mediator, the decolorization of Reactive orange 16 and Remazol brilliant blue R was also achieved.

Rabbit Model for in vivo Study of Intervertebral Disc Degeneration and Regeneration

  • Kong, Min-Ho;Do, Duc-H.;Miyazaki, Masashi;Wei, Feng;Yoon, Sung-Hwan;Wang, Jeffrey C.
    • Journal of Korean Neurosurgical Society
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    • v.44 no.5
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    • pp.327-333
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    • 2008
  • Objective: The purpose of this study is to verify the usefulness of the rabbit model for disc degeneration study. Materials: The L1-L2, L2-L3, L3-L4. or L4-L5 lumbar intervertebral disc (IVD) of 9 mature male New Zealand White rabbits were injured by inserting a 16-gauge needle to a depth of 5 mm in the left anterolateral annulus fibrosus while leaving L5-L6 IVD uninjured. Three other rabbits also received intradiscal injections of rabbit disc cells transfected with adenovirus and bone morphogenetic protein-2 (ad-BMP-2) at L4-L5 in addition to injury by 16-gauge needle at the L1-L2 level. Using digitized radiographs, measurements of IVD height were made and analyzed by using the disc height index (DHI). Magnetic resonance imaging (MRI) scans of the injured discs, injected discs, and uninjured L5-L6 discs were performed at 15 weeks post surgery and compared with preoperative MRI scans. Results: All twelve rabbits showed consistent results of disc degeneration within 15 weeks following annular puncture. DHIs of injured discs were significantly lower than that of the uninjured L5-L6 discs (p<0.05). The mean value of disc degeneration grade of injured discs was significantly higher than that of uninjured discs (p<0.05). The injection of disc cell transfected with ad-BMP-2 did not induce disc regeneration at 15 weeks after injection. Conclusion: This study showed that the injured disc had a significant change in DHI on simple lateral radiograph and disc degeneration grade on MRI scans within 15 weeks in all rabbits. Rabbit annular puncture model can be useful as a disc degeneration model in vivo.

The Analysis of Geriatric Nutritional Risk Index (GNRI) for Nutritional Assessment and Health Care in Elderly Women (여자노인의 영양상태 평가 및 건강관리를 위한 Nutritional Risk Index (NRI) 비교 분석)

  • Yang, Eun-Ju
    • Journal of Nutrition and Health
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    • v.42 no.3
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    • pp.234-245
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    • 2009
  • Nutritional assessment for the elderly can identify health status and morbidity. However, development of Nutritional Risk Index (NRI) remains limited for elderly because of difficulties in understanding physiological mechanism of elderly. This study was performed to analyze and develop Nutritional Risk Index for Korean elderly Women (Geriatric Nutritional Risk Index, GNRI). Based on literature review, factors for NRI were identified and indices were assessed by a cross-sectional survey. The survey involved Korean elderly women (${\geq}$60, n = 94) in Gwangju area, and sociodemographics, lifestyle characteristics, health conditions, dietary intakes based on 24h- recall, anthropometric measures (wt, ht, BMI, waist, hip, WHR, body protein, body fat, abdominal fat, and triceps skinfold thickness), and clinical biochemistry parameters (systolic blood pressure, diastolic blood pressure, cholesterol, HDL-cholesterol, triglyceride, total protein, albumin, prealbumin, hemoglobin, hematocrit, fasting blood glucose, HbAlc, ferritin, Zn, Ca, Na, K, Vit E, Vit $B_{12}$, folate, C-reactive protein) were examined relation to nutritional risk index. Based on literature review and data analyses, three NRIs were categorized (NRI I, NRI II, NRI III) and used for further analysis. NRI I was related to having metabolic syndrome, NRI II was related to serum albumin and body weight, and NRI III was related to food habit and health concerns. Abdominal fat (%) of elderly was correlated with each NRIs. NRI II was correlated with nutritional deficiency and higher tendency of inflammatory response, and NRI III was correlated with nutritional status which tend to be lower on aging (protein, folate, Vit $B_{12}$). NRI can serve as a useful tools in assessing health risk and nutritional status. Some modification of items in NRI and validity study are need to apply to Korean elderly.

Studies on the Antioxidative Substances in the Seeds of the Rutaceae Family (운향과(芸香科) 식물(植物) 종실(種實)의 항성화성(抗醒化性))

  • Kim, Seong-Jin;Kim, Ji-Soo;Joh, Yong-Goe
    • Journal of the Korean Applied Science and Technology
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    • v.11 no.1
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    • pp.7-16
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    • 1994
  • Some seeds of the Rutaceae family, Zanthoxylum piperitum, Z. schinifolium officinalis, Poncirus trifoliata, Citrus unshin, were investigated to clarify their antioxidative components. Finely powdered samples were extracted by hexane, followed by dichioromethane and then 70% methanol in a hot bath. Its unsaponifiables containing X-and Y-tocopherol with trace amount of ${\beta}-and$\;{\delta}-tocopherol$. also showed comparatively weak activity, although the hexane fraction itself had no significant antioxidative effect on lard. Levels of total tocopherols in the samples averages 42. 24-154. 11 mg/lOOg total extractives. The dichloromethane-and 70% methanol extractives showed strong antioxidative activity, from which antioxidative substances were purified with benzene-acetone(6:5, V/V) on a silica gel column, and with a solvent mixture of acetonitrile-methanol-$H_2O$(40:40:20, V/V/V) on a Sep-Pak $C_{18}$ hydrolyzed by 5% KOH-ethanol. The recovered unsaponifiables were, then, separated on a column of high performance liquid chromatography. The unsaponifiables produced by hydrolysis of the isolates from dichloromethane extractives has epi-catechin(40.0-57.1%) and (+)-catechin<$l9.1{\sim}24.4%$ to total phenolic substances, on area base) as major component, accompanied by chlorogenic acid, gallic acid(?), trans-p-coumaric acid and tralls-p-ferulic acid including some unknown components, and those derived from 70% methanol extractives also comprise (+)-catechin($31.3{\sim}39.6%$ to total components, on area base), epi-catechin($2O.2{\sim}36.4%$), trans-p-cournaric acid(8.4-15.3%) and trans-p-ferulic acid($7.7{\sim}14.1%$) as predominant component with some minor coponents, but the fraction supposed to be gallic acid(?) is not present. The antioxidative activities of the phenolic components isolated in this work were in order of epi-catechin>catechin>chlorogenic acid>trans-p-ferluic acid>trans-p-coumaric acid.

The Proteinase Distributed in the Intestinal Organs of Fish 3. Purification and Some Enzymatic Properties of the Alkaline Proteinases from the Pyloric Caeca of Skipjack, Katsuwonus vagans

  • PYEUN Jae-Hyeung;KIM Hyeung-Rak;HEU Min-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.21 no.2
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    • pp.85-96
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    • 1988
  • Purification and some properties of alkaline proteinases in the pyloric caeca of skipjack, Katsuwonus vagans, were investigated. Four alkaline proteinases, temporarily designated proteinases I, II, III and IV, were identified from the tissue extract of the pyloric caeca by ammonium sulfate fractionation, DEAE-Sephadex A-50 chromatography, and Sephadex G-100 and G-200 gel filtration. Result of disc-polyacrylamide gel electrophoretic analysis showed that the purified proteinases II and III were homogenous with the yields of $1.5\%\;and\;1.2\%$, and those specific activities were increased to 33 to 37 fold over that of the crude enzyme solution, respectively. Molecular weight of the proteinases II and III determined by sephadex G-100 gel filtration were 28,500 and 24,200, respectively. The optimum conditions for the caseinolytic activity of the two enzymes were pH 9.6 and $48^{\circ}C$. The reaction rates of the two alkaline proteinases were constant to the reaction time to 80 min in the reaction mixture of $3.4{\mu}g/ml$ of enzyme concentration and $2\%$ casein solution. The Km values against casein substrate determined by the method of Lineweaver-Burk were $0.56\%$ for proteinase II and $0.30\%$ for proteinase II. The proteinases II and III were inactivated under the presence of $Ag^+,\;Hg^{2+},\;Ni{2+},\;Fe^{2+},\;and\;Cu^{2+}$, and but activated by $Mn^{2+}\;and\;Ca^{2+}$ and markedly inhibited by the soybean trypsin inhibitor and N-p-toluenesulfonyl-L-lysine chloromethyl ketone. Therefore, the proteinases II and III were found to be a group of serine proteases and assured to be trypsin-like proteinases.

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A Novel Ginsenosidase from an Aspergillus Strain Hydrolyzing 6-O-Multi-Glycosides of Protopanaxatriol-Type Ginsenosides, Named Ginsenosidase Type IV

  • Wang, Dong-Ming;Yu, Hong-Shan;Song, Jian-Guo;Xu, Yu-Feng;Liu, Chun-Ying;Jin, Feng-Xie
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1057-1063
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    • 2011
  • Herein, a novel ginsenosidase, named ginsenosidase type IV, hydrolyzing 6-O-multi-glycosides of protopanaxatriol-type ginsenosides (PPT), such as Re, R1, Rf, and Rg2, was isolated from the Aspergillus sp. 39g strain, purified, and characterized. Ginsenosidase type IV was able to hydrolyze the 6-O-${\alpha}$-L-($1{\rightarrow}2$)-rhamnoside of Re and the 6-O-${\beta}$-D-($1{\rightarrow}2$)-xyloside of R1 into ginsenoside Rg1. Subsequently, it could hydrolyze the 6-O-${\beta}$-D-glucoside of Rg1 into F1. Similarly, it was able to hydrolyze the 6-O-$_{\alpha}$-L-($1{\rightarrow}2$)-rhamnoside of Rg2 and the 6-O-${\beta}$-D-($1{\rightarrow}2$)-glucoside of Rf into Rh1, and then further hydrolyze Rh1 into its aglycone. However, ginsenosidase type IV could not hydrolyze the 3-O- or 20-O-glycosides of protopanaxadiol-type ginsenosides (PPD), such as Rb1, Rb2, Rb3, Rc, and Rd. These exhibited properties are significantly different from those of glycosidases described in Enzyme Nomenclature by the NC-IUBMB. The optimal temperature and pH for ginsenosidase type IV were $40^{\circ}C$ and 6.0, respectively. The activity of ginsenosidase type IV was slightly improved by the $Mg^{2+}$ ion, and inhibited by $Cu^{2+}$ and $Fe^{2+}$ ions. The molecular mass of the enzyme, based on SDS-PAGE, was noted as being approximately 56 kDa.

Miniaturized CPW-fed Folded Slot Antenna (소형화된 CPW 급전 폴디드 슬롯 안테나)

  • Woo, Hee-Sung;Shin, Dong-Gi;Lee, Young-Soon
    • Journal of Advanced Navigation Technology
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    • v.24 no.2
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    • pp.142-147
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    • 2020
  • In the present study, we proposed newly a CPW-fed miniaturized folded-slot antenna with open ended slot for WCDMA (1.92 ~ 2.17 GHz) band. Open-ended slots and asymmetric ground plane are used for a miniaturization of the antenna, and the proposed antenna was designed and fabricated on a FR-4 substrate with dielectric constant 4.3, thickness of 1.6 mm, and size of 35×70 ㎟. The measured impedance bandwidths (|S11| ≤ -10 dB) of fabricated antenna is about 400 MHz (1.86 ~ 2.26 GHz), which sufficiently satisfied interested band. Furthermore, the gain of antenna is 2 dBi and this antenna shows a similar radiation patterns of the dipole antenna. Therefore, it is expected to be used usefully in wireless and mobile communication device.

Effect of growth temperature on the growth and properties of carbon-nanotube prepared by Hot-filamnet PECVD method (Hot-filament 화학기상 증착법으로 성장시킨 성장온도에 따른 탄소나노튜브의 성장 및 특성)

  • Kim, Jung-Tae;Park, Yong-Seob;Kim, Hyung-Jin;Lee, Sung-Uk;Choi, Eun-Chang;Hong, Byung-You
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2006.11a
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    • pp.120-120
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    • 2006
  • 탄소나노튜브는 nm급의 크기에 높은 전기전도도, 열전도 효율, 감한 기계적 강도 등의 장점을 가지며, FED(Field Emission Display), 극미세 전자 스위칭 소자, SET(Single Electron Transistor), AFM(Atomic Force Microscope) tip등 여러 분야로의 응용을 연구하고 있다. 본 연구에서는 탄소나노튜브를 Si 웨이퍼 위에 Ni/Ti 금속층을 촉매층으로 사용하고, 암모니아($NH_3$)가스와 아세틸렌 ($C_2H_2$)가스를 각각 희석가스와 성장원으로 사용하여 합성하였다. 탄소나노튜브의 성장은 Hot filament 화학기상증측(HFPECVD) 방식을 사용하였으며, 이 방법은 다량의 합성, 높은 균일성, 좋은 정렬 특성등의 장점을 가진다. 성장 온도는 탄소나노튜브의 성장 특성을 변화시키는 중요한 요소이다. 성장 온도에 따라 수직적 성장, 성장 밀도등의 특성 변화를 관찰하였다. 성장된 탄소나노튜브층 성분 분석은 에너지 분산형 X-선 측정기(EDS)를 통해 관찰하였고, 끝단에 촉매층이 존재하는 30~50 nm 폭을 가진 다중벽 탄소나노튜브를 고배율 투과전자현미경(HRTEM) 분석을 통해 관찰하였다. 전계방사 주사전자현미경(FESEM) 분석을 동해 1~3${\mu}m$의 길이를 가진 탄소나노튜브가 높은 밀도로 성장된 것을 확인하였다.

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Promoting Effect of MgO in the Photodegradation of Methylene Blue Over MgO/MWCNT/TiO2 Photocatalyst

  • Chen, Ming-Liang;Zhang, Feng-Jun;Oh, Won-Chun
    • Korean Journal of Materials Research
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    • v.20 no.7
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    • pp.345-350
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    • 2010
  • For the present paper, we prepared MgO/MWCNT/$TiO_2$ photocatalyst by using multi-walled carbon nanotubes (MWCNTs) pre-oxidized by m-chlorperbenzoic acid (MCPBA) with magnesium acetate tetrahydrate $(Mg(CH_2COO)_2\cdot4H_2O)$ and titanium n-butoxide $(Ti\{OC(CH_3)_3\}_4)$ as magnesium and titanium precursors. The prepared photocatalyst was analyzed by X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) analysis. The decomposition of methylene blue (MB) solution was determined under irradiation of ultraviolet (UV) light. The XRD results show that the MgO/MWCNT/$TiO_2$ photocatalyst have cubic MgO structure and anatase $TiO_2$ structure. The porous structure and the $TiO_2$ agglomerate coated on the MgO/MWCNT composite can be observed in SEM images. The Mg, O, Ti and C elements can be also observed in MgO/MWCNT/$TiO_2$ photocatalyst from EDX results. The results of photodegradation of MB solution under UV light show that the concentration of MB solution decreased with an increase of UV irradiation time for all of the samples. Also, the MgO/MWCNT/$TiO_2$ photocatalyst has the best photocatalytic activity among these samples. It can be considered that the MgO/MWCNT/$TiO_2$ photocatalyst had a combined effect, the effect of MWCNT, which could absorb UV light to create photoinduced electrons $(e^-)$, and the electron trapping effect of MgO, which resulted in an increase of the photocatalytic activity of $TiO_2$.

Synthesis and Photocatalytic Activity of Ag Spot-coated TiO2-SrO Composite Powders (나노 Ag spot-coated TiO2-SrO 복합분체의 합성과 광촉매 활성 평가)

  • Han, Jae-Kil;Kim, Hyeong-Chul;Hong, Won-Seok;Choi, Sung-Chang
    • Korean Journal of Materials Research
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    • v.18 no.7
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    • pp.389-393
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    • 2008
  • Nano-sized $TiO_2$-60 wt% SrO composite powders were synthesized by a sol-gel method using titanium isopropoxide and $Sr(OH)_2\;{\cdot}\;8H_2O$ as precursors. 3, -5, -7 wt%Ag spot-coated $TiO_2$-60 wt% SrO composite powders were synthesized by a Ag electroless deposition method using $TiO_2$-60 wt% SrO composite powders calcined at $1050^{\circ}C$, which mainly exhibited the $SrTiO_3$phase. However, a small number of rutile $TiO_2$, $Sr_2TiO_4$ and $SrO_2$ phases were also detected. In the Ag spot-coated powders synthesized by electroless deposition, nano-sized particles about 5-25 nm in diameter adhered to the $TiO_2$-60 wt% SrO composite powders. The photocatalytic activity of Ag spot-coated $TiO_2$-SrO and $TiO_2$-SrO composite powders for degradation of phenol showed that all of $TiO_2$-SrO composite powders were highly active under UV light irradiation. 7 wt%Ag spot-coated $TiO_2$-60wt.%SrO composite powders had a relatively higher photocatalytic activity than did $TiO_2$-SrO composite powders under visible light.