• Title/Summary/Keyword: natural purification

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Purification and Characterization of $\beta$-Cyclodextrin Glucanotransferase Excreted by Bacillus firmus var. aikalophilus. (호알칼리성 Bacillus firmus가 생산하는 $\beta$-Cyclodextrin Glucanotransferase의 정제 및 효소반응 특성)

  • Shin, Hyun-Dong;Kim, Chan;Lee, Yong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.323-330
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    • 1998
  • Cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of the Bacillus firmus var. alkalophilus, using ultrafiltration, starch adsorption/desorption, ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephacryl HR-100. The molecular weight of the purified enzyme was determined as 77,000 by SDS-PAGE. The optimum pH and temperature for the CD synthesis were 6.0 and 5$0^{\circ}C$, respectively. The activity of this enzyme was stably kept at the range of pH 6.0~9.5 and up to 5$0^{\circ}C$. However, in the presence of $Ca^{2+}$, the optimum temperature for CD synthesis was shifted 55~6$0^{\circ}C$ and this enzyme was stable up to 6$0^{\circ}C$ because of the stabilizing effect of $Ca^{2+}$. The purified CGTase produced CDs with high conversion yields of 45~51% from sweet potato starch, com starch and amylopectin as substrate, especially, and the product ratio of $\beta$-CD to ${\gamma}$-CD was obtained at range of from 5.8:1 to 8.4:1 according to the kind of substrate. The purified enzyme produced mainly $\beta$-CD without accumulation of $\alpha$-CD during enzyme reaction using various starches as the substrate, indicating that the purified enzyme is the typical $\beta$-CGTase. The purified CGTase produced 25 g/l of CDs from 5.0% (w/v) liquefied com starch and the conversion yield of CDs was 50%, and the content of $\beta$-CD was 84% of total CDs after 8 hours under the optimum reaction condition.ion.

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Physicochemical properties of deposited particles on surface of pine leaves as biomarker for air pollution (솔잎가지 표면에 침착된 입자상 물질의 물리화학적 특성 및 대기오염 지표로서의 가능성 고찰)

  • Chung, David;Choi, Jeong-Heui;Lee, Jang-Ho;Lee, Soo-Yong;Lee, Ha-Eun;Park, Ki-Wan;Shim, Kyu-Young;Lee, Jong-Chun
    • Analytical Science and Technology
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    • v.31 no.6
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    • pp.247-258
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    • 2018
  • The purpose of the present study was to investigate whether the degree of air pollution can be evaluated via examination of local plants. Selected sites included two parks in an industrial area, as well as two parks in an urban area. Selected plant samples comprised one-year-old pine shoot leaves. Leaves growing over 2 m from the ground were collected from over 10 pine trees. Leaf surface was analyzed for deposition of 14 trace elements and 16 polycyclic aromatic hydrocarbons (PAHs), including particle size and mass, surface imaging, precipitation-mediated particle removal rate, and concentration. Particle size ranged from 0.4 to $200{\mu}m$, and the volume percentage of particles ${\leq}10$ was 20 %. Deposited particle mass ranged from 0.450-0.825 mg, and precipitation-mediated removal rate ranged from 10.0-27.6 %. Trace element concentration, as measured by ICP/MS after microwave acid digestion, was 18.8-26.3 mg/kg As, 0.08-0.13 mg/kg Be, 0.06-0.08 mg/kg Cd, 4.91-17.8 mg/kg Cr, 5.26-405 mg/kg Cu, 1,930-2,670 mg/kg Fe, 3.03-28.1 mg/kg Pb, 26.9-42.8 mg/kg Mn, 2.66-10.4 mg/kg Ni, 4,560-8,730 mg/kg Al, 2,500-6,120 mg/kg Ba, 5.27-17.8 mg/kg Rb, 40.9-95.3 mg/kg Sr, and 4,030-8,260 mg/kg Zn. Concentration of PAHs, as analyzed by GC/MS/MS after liquid-liquid extraction and purification of deposited particles, ranged from 1.17 to 12.378 mg/kg for ${\Sigma}PAH_{16}$ and from 1.17 to 12.378 mg/kg for ${\Sigma}PAH_7$.

A Study on the Natural Insectifuge for Food Wrapping Corrugated Board Using Tree Extractives (수목 추출성분을 이용한 식품포장용 골판지 천연 방충처리제 개발)

  • 배영수
    • Journal of Korea Foresty Energy
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    • v.20 no.2
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    • pp.9-19
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    • 2001
  • This study was carried out to investigate natural insectifuge materials from tree extractives in order to substitute for organic synthetic insecticides for food wrapping corrugated board. Tree samples were collected, extracted, fractionated with hexane, $CH_2Cl_2$, ethylacetate(EtOAc) and $H_2O$, and then freezed dried for further study. EtOAc or $H_2O$ fractions were chromatographed on a Sephadex LH-20 column for isolation and purification, and the isolated compounds were characterized by spectroscopic tools such as NMR and MS. Crude extractives of EtOAc and $H_2O$ fractions were added to the printing ink for corrugated board with the concentration of 2% or 3% based on the weight of the ink, then the prepared ink was printed on the corrugated board to be used for evasion test using larva of indian meal moth(Plodia interpunctella(Hubner)). Robtin, dihydrorobinetin and leucorobinetinidin were isolated from the wood extractives of black locust(Robinia pseudoacacia) and the bark of poplar(Populus alba $\times$ glandulosa) contained many kinds of compounds such as (+)-catechin, naringenin, aromadendrin, eriodictyol, sakuranetin and its glycoside, taxifolin, neosaturanin, salireposide, p-coumaric acid and aesculin. Much of (+)-catechin was isolated from the bark extractives of willow(Salix koreensis) in addition to (+)-gallocatechin and p-coumaric acid and the bark of weeping willow(Salix babylonica) also contained (+)-catechin, (+)-gallocatechin, dihydromyricetin and myricetin.

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Purification and Characterization of ($Ca^{2+}$+$Mg^{2+}$)-ATPase of Sarcoplasmic Reticulum from Rat Skeletal Muscle (쥐 근소포체의 ($Ca^{2+}$+$Mg^{2+}$)-ATPase의 분리정제와 그 효소특성에 관하여)

  • Lee, Jong-Soon;Ha, Doo-Bong;Chung, Chin-Ha
    • The Korean Journal of Zoology
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    • v.28 no.1
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    • pp.31-43
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    • 1985
  • The $(Ca^{2+}+Mg^{2+})$-ATPase has been purified homogeneously from sarcoplasmic reticulum of rat skeletal muscle by sucrose density gradient centrifugation. The purified enzyme has a molecular weight of 115,000 as judged by polyacrylamide gel electrophoresis in the presence of sodium dedecyl sulfate, and therefore has the same size of the enzyme in rabbit and chick skeletal muscle. $Ca^{2+}, Mg^{2+}, Fe^{2+}, Co^{2+}, and Mn^{2+}$ at 50 $\\muM$ show stimulatory effect on the ATP-ase, while $Zn^{2+}, Cu^{2+}, and Hg^{2+}$ inhibit it at the same concentration. The ATPase activity is insensitive to antimalarial drugs such as quinine and quinacrine, but is sensitive to inhibition by p-hydroxymecurie benzoate and phenylmethylsulfonylfluoride. The enzyme has optimum pH of 6 to 7 and Km value for ATP is estimated to be 98 $\\muM$. Thus, a number of biochemical properties of this enzyme appear to be different from those of the enzyme that have been isolated from rabbit skeletal muscle. The $(Ca^{2+}+Mg^{2+})$-ATPase appears to be selectively degraded in microsomal fraction. The activity of metalloendoprotease is evident in the microsomal preparation when assayed by radioactively labeled protein substrate, such as $^{3}H-casein and $^{125}I$-insulin. However, it is presently unclear whether the metalloendoprotease is responsible for the degradation of the $(Ca^{2+}+Mg^{2+})$-ATPase.

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Immunogenicity of Synthetic Peptide Specific for Major Immunogenic Determinat of Hepatitis B Surface Antigen (B형간염(型肝炎) 표면항원(表面抗原)의 주면역원(主免疫原) 결정기(決定基)에 특이(特異)한 합성(合成) Peptide의 면역원성(免疫原性)에 관한 연구(硏究))

  • Shin, Kwang-soon;Han, Su-nam
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.7-17
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    • 1985
  • Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

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A Study on Efficiency of Water Purification of Korean Village Bangjuk[dike] as a Means of Ecological Watershed Management (생태적 유역관리 도구로써 마을방죽의 수질정화 효율성 고찰)

  • An, Byung-Chul
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.30 no.2
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    • pp.90-100
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    • 2012
  • This study centering on 10 village - Bangjuks analyzed multifunctionality value of village Bangjuks which have been main water treatment system in Korean traditional villages. On the basis of understanding the structure and character of components such as the well, common spring, village waterway and others which making water-flow and consisting of aquatic system in Korean traditional village Bangjuk, the conclusion as the instrumental device of social and ecological role and ecological watershed management, securing the ecosystem soundness of the damaged or deteriated aquatic ecosystem due to the industrialization and urbanization is as below; 1. The traditional village Bangjuk was environmentally friendly hydraulic system which gathers waterways of village into a point including sewage water, retains and flows out to village through agricultural waterway. Through this Bangjuk, they have managed sewage and rainfall runoff flowed out village efficiently. It is not only a detention system of water but a kind of eco-friendly system that flow out water into the rivers after reusing and filtering it. 2. Around five traditional villages and five villages after modernization, this study classified the types of village Bangjuk as three types considering geographic location, size, etc; marsh type of low swamp, high water -low rice field type of natural flow stucture, low water - high rice field type requiring artificial irrigation facility. All the five traditional villages were turned out to be marsh type of low swamp. Geoji, Sanjeri, Ma-am, Yangchon of the agricultural villages were high water-low rice filed type, and Sangchoenri village was classified low water-high rice field type. 3. This study checked up the function of water purification of village Bangjuk. In Wonteo and Geji villages affected by discharge of village sewer and domestic sewage, the efficiency of ammonia nitrogen($NH_3-N$) and total phosphorus(T-P) was 56~95%, which was high. In Sangcheonri and Sanjeri villages strongly affected by stall and farmland, the efficiency of suspended solids(SS) was 70~85%, and that of total nitrogen(T-N) and total phosphorus(T-P) was 5.3~65%. 4. A water purification system can be found out in the system of village Bangjuk that filter out village sewage and rainfall runoff flowed through the settle and filter of pollution source and denitrification of plants. Through this system of village Bangjuk, it must be used as the basic facilities for the ecological watershed management. The sewage management system of village Bangjuk as a eco-filter must be used and studied as an eco-friendly facility for the ecological watershed management around the subwatershed and catchment.

Inhibitory Activity of Medicinal Herbs on Nitric Oxide Synthesis in Activated Macrophages

  • Lee, Hwa-Jin;Kim, Ji-Sun;Jin, Chang-Bae;Ryu, Jae-Ha
    • Natural Product Sciences
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    • v.11 no.1
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    • pp.16-21
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    • 2005
  • Nitric Oxide (NO), derived from L-arginine, is produced by two types (constitutive and inducible) of nitric oxide synthase (NOS: cNOS and iNOS). The NO produced in large amounts by the iNOS is known to be responsible for the vasodilation and hypotension observed in septic shock, cancer metastasis and inflammation. The inhibitors of iNOS, thus, may be useful candidates for the treatment of inflammatory diseases accompanied by the overproduction of NO. We prepared alcoholic extracts of herbal drugs which have been used for the treatment of inflammation in oriental medicine. We have screened the inhibitory activity of NO production in lipopolysaccharide (LPS)-activated macrophages after the treatment of these extracts. Among 82 kinds of extracts of herbal drugs, 35 extracts showed the potent inhibitory activity of NO production above 50% at the concentration of $50\;{\mu}g/mL$. The inhibitory activities of NO production were also evaluated for several solvent fractions at two different concentrations. Especially, hexane and EtOAc fractions of Alpinia officinarum, Angelica gigas, Ostericum koreanum, Saussurea lappa, Torilis japonica, and hexane fractions of Agrimonia pilosa, Machilus thunbergii, Hydrangea serrata, Magnolia obovata, Prunella vulgaris, Tussilago farfara, and EtOAC fractions of Perilla frutescence showed a significant activity at 10 and/or $25\;{\mu}g/mL$. In Western blot analysis, the hexane fractions ($5\;{\mu}g/mL$) of Magnolia obovata and Saussurea lappa, and EtOAc fractions ($20\;{\mu}g/mL$) of Hydrangea Serrata, Perilla frutescence and Torilis japonica inhibited the expression of iNOS protein in LPS-activated macrophages. These plants may be promising candidates for the study of the activity-guided purification of active compounds and might be useful for the treatment of inflammatory diseases and endotoxemia accompanying overproduction of NO.

Effect of Coagulated Flocs Broken by the Pressure Pump on Removal Rate and Membrane Fouling of Pressurized MF process (가압펌프에 의해 해체된 플럭이 가압식 막여과 공정의 제거효율 및 막오염에 미치는 영향)

  • Kim, Junhyun;Moon, Baeksu;Park, Jongsu;Cho, Yoonho;Kim, Jinho
    • Membrane Journal
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    • v.23 no.6
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    • pp.460-468
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    • 2013
  • This study reviewed optimum dosage rate of coagulant and ability to remove dissolved organic carbon without sedimentation in conventional water purification plant. It was confirmed that floc formated by pre-treatment process was broken by impeller of booster pump. Optimum dosage rate of coagulant was 4 mg/L (as PACl 17%) for floc formation through blend, coagulation and after passing through the pump when turbidity of raw water was less than 10 NTU. And average removal rate of dissolved organic carbon was 43% at that time. Maximum removal rate of dissolved organic carbon was 48%, even though coagulation rate was increased gradually until 8 mg/L (as PACl 17%). So removal rate of dissolved organic carbon is not much improved even if dosage rage of coagulant increase. TMP of PVDF (polyvinylidene flouride) pressurized MF process without pre-treatment operated at 0.54 bar and TMP of PVDF pressurized MF process with pre-treatment operated at 0.41 bar.

Design for Landfill Gas Application by Low Calorific Gas Turbine and Green House Optimization Technology (Low Calorific Gasturbine 매립지 적용 및 유리온실 운용기술 설계)

  • Hur, Kwang-Beom;Park, Jung-Keuk;Lee, Jung-Bin;Rhim, Sang-Gyu
    • 한국신재생에너지학회:학술대회논문집
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    • 2010.06a
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    • pp.244.1-244.1
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    • 2010
  • Bio energy development by using Low Calorific Gas Turbine(LCGT) has been developed for New & Renewable energy source for next generation power system, low fuel and operating cost method by using the renewable energy source in landfill gas (LFG), Food Waste, water waste and Livestock biogas. Low calorific fuel purification by pretreatment system and carbon dioxide fixation by green house system are very important design target for evaluate optimum applications for bio energy. Main problems and accidents of Low Calorific Gas Turbine system was derived from bio fuel condition such as hydro sulfide concentration, siloxane level, moisture concentration and so on. Even if the quality of the bio fuel is not better than natural gas, LCGT system has the various fuel range and environmental friendly power system. The mechanical characterisitics of LCGT system is a high total efficiency (>70%), wide range of output power (30kW - 30MW class) and very clean emmission from power system (low NOx). Also, we can use co-generation system. A green house designed for four different carbon dioxide concentration from ambient air to 2000 ppm by utilizing the exhaust gas and hot water from LCGT system. We look forward to contribute the policy for Renewable Portfolio Standards(RPS) by using LCGT power system.

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Neuroprotective and Anti-Neuroinflammatory Activities of Anthraquinones Isolated from Photorhabdus temperata Culture Broth

  • Yang, Eun-Ju;Kim, Seo-Hyun;Lee, Kyeong-Yeoll;Song, Kyung-Sik
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.12-21
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    • 2018
  • Photorhabdus temperata (PT), a gram-negative bacterium, lives symbiotically within entomopathogenic nematodes. The insecticidal compounds derived from Photorhabdus are used as biopesticides in agriculture. However, the physiological properties are not well characterized. In the course of our screening for neuroprotective and anti-neuroinflammatory substances from natural products, the culture broth of PT showed considerable activities. By activity-guided purification, five anthraquinones, namely, 3-methoxychrysazine (1), 1,3-dimethoxy-8-hydroxy-9,10-anthraquinone (2), 1,3,8-trihydroxy-9,10-anthraquinone (3), 3,8-dihydroxy-1-methoxy-9,10-anthraquinone (4), and 1,3,4-trimethoxy-8-hydroxy-9,10-anthraquinone (5), were isolated from the ethyl acetate fraction of the PT culture broth. Among the isolated compounds, $75{\mu}M$ 3 significantly protected mouse hippocampal neuronal cells (HT22) against 5 mM glutamate-induced cell death via the inhibition of reactive oxygen species production, $Ca^{2+}$ influx, and lipid peroxidation. Additionally, 3 and 4 effectively suppressed the interferon-${\gamma}$-induced neuroinflammation of mouse-derived microglial cells (BV2) at 10 ng/ml, via the reduction of nitric oxide, interleukin-6, and tumor necrosis factor-${\alpha}$. Anthraquinones 3 and 4 derived from the PT culture broth are a potential starting point to discover neuroprotective and anti-neuroinflammatory drug leads. The novel compound 5 is reported for the first time in this study.