• Title/Summary/Keyword: Purification plant

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Purification of Cucurbitacins D, E, and I from Ecballium Elaterium (L.) A. Rich Fruits and Study of Their Cytotoxic Effects on the AGS Cell Line

  • Jafargholizadeh, Naser;Zargar, Seyed Jalal;Yassa, Narguess;Tavakoli, Saeed
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.10
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    • pp.4631-4635
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    • 2016
  • Background: The plant Ecballium elaterium (L.) A. Rich, belongs to the Cucurbitaceae family which occupies an important position in traditional medicine prescriptions. It has been reported that a freeze-dried aqueous extract of E. elaterium fruits has cytotoxic effects on the AGS human stomach adenocarcinoma cell line. We here focused on anticancer effects of the main chemicals purified from E. elaterium fruits. Materials and Methods: We isolated cucurbitacins D, E, and I from chloroform, and ethyl acetate fractions of a methanolic extract of E. elaterium fruits and assessed their cytotoxic effects on the AGS cell line by MTT assay. The methanolic extract was fractionated to petroleum ether, chloroform, and ethyl acetate fractions. The compounds isolated by column chromatography were identified by NMR spectroscopy. Results: After 24 h of incubation with AGS cells, the IC50 values were 0.3, 0.1, and $0.5{\mu}g/ml$ for cucurbitacins D, E, and I respectively. Conclusions: This finding suggests that because of its cucurbitacins, E. elaterium fruit may have some cytotoxic effects on gastric cancer cells. Also, compared with D and I, cucurbitacin E showed greater potency in this regard.

Brassinosteroid substances in immature oryza sativa seeds (벼종자의 brassinosteroid 활성물질)

  • Park, Keun-Hyung;Kim, Seon-Jae;Park, Jong-Dae;Lee, Lan-Sook;Hyun, Kyu-Hawn
    • Applied Biological Chemistry
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    • v.36 no.5
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    • pp.376-380
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    • 1993
  • To investigate the presence of the brassinosteroid substances in immature Oryza sativa L. cv Tongjinbyeo seeds, the methanol extract was purified by the sequential use of solvent fractionation, silica gel adsorption chromatography, Sephadex LH-20 chromatography and charcoal adsorption chromatography. The activity of brassinosteroid was monitored by the rice inclination test and its presence could be confirmed in each purification step. The purified active components were separated by silica gel adsorption chromatography and Sephadex LH-20 chromatography. Brassinosteroid substances in separated active fractions were identified as castasterone, teasterone and 6-deoxocastasterone by HPLC. Our work is probably the first report of endogenous brassinosteroids in Oryza sativa seeds. The content of brassinosteroid in Oryza sativa seeds as converted into brassinolide was $0.5{\sim}1.5\;ng/g$ fresh weight.

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Purification and Characterization of a Novel Antifungal Protein from Paenibacillus macerans PM1 Antagonistic to Rice Blast Fungus, Pyricularia oryzae

  • Bae, Dong-Won;Kawk, Weon-Sik;Lee, Joon-Taek;Son, Dae-Young;Chun, Sung-Sik;Kim, Hee-Kyu
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.805-810
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    • 2000
  • An antifungal protein antagonistic to the rice blast fungus, Pyricularia oryzae was purified from Paenibacillus macerans PM-1 by ammonium sulfate fractionation, Q Sepharose Fast Flow column chromatography, Phenyl Sepharose CL-4B column chromatography and Superose 12 gen filtration. An apparent molecular mass of the purified antifungal protein was determined as 8 kDa by SDS-PAGE and 9 kDa by analytical gel filtration, respectively, suggesting that the purified protein is a monomer. The antifungal protein was stable at pH range from 7-12 and up to $100^{\circ}C$. The protein was also stable at 0.1-1% Tween 20 and Triton X-100. The N-terminal amino acid sequence of the antifungal protein was Thr-Glu-Leu-Pro-Leu-Gly-Ile-Val-Met-Asp-Lys-Tyr-Thr-Asp-Ala-Phe-Lys-Phe-Asp-Met-Phe. Comparison of the determined sequence with other peptide and DNA sequences did not reveal homology at all. Therefore, the purified antifungal protein was speculated to be a novel protein. The condidial germination in vitro of P. oryzae KJ301:93-39 by the purified protein ($5.9{\mu} g/ml$) was limited to $9{\pm}3.2%$ only, compared with $69{\pm}2.4%$ of the control. Ungerminated conidia were swollen at basa and mid cell by the purified protein. In vivo bioassay for inhibition of conidial germination of P. oryzae KJ 301, one of the most predominating racesin Korea. the purified protein ($5.9{\mu} g/ml$)strongly inhibited the conidial germination. The conidia, even though germinated, could not develop any further to produce appressoria efficiently.

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A Feasibility Study for Renewable Energy from Sewage Sludge Biogas (하수슬러지 Biogas의 신재생에너지화 타당성 연구)

  • Kang, Ho;Lee, Hye Mi;Cho, Sang Sun;Park, Sun Uk;Jeong, Ji Hyun
    • Journal of Korean Society on Water Environment
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    • v.26 no.5
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    • pp.754-760
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    • 2010
  • This study was carried out not only to evaluate optimal operating condition to increase biogas production, but also to estimate feasibility of renewable energy from anaerobic digester of sewage sludge. Semi- continuous Fed and Mixed Reactors (SCFMRs) were operated in various condition to quantify the reactor variables. The result of SCFMR operation showed that the biogas productivity and total volatile solids (TVS) removal of total solids (TS) 4% reactor at hydraulic retention time (HRT) 20 days with Organic Loading Rate (OLR) of $1.45kg/m^3-d$ were $0.39m^3/m^3-d$ and 26.7%, respectively which was two times higher than that of TS 2.5% reactor. Consequently the daily biogas production of $20,000m^3$ would be possible from the total volume of $52,000m^3$ of anaerobic digesters of the municipal wastewater treatment plant in D city. In feasibility study for the Biogas utilization, combined heat and power system (CHP) and CNG gasification were examined. In case of CHP, the withdrawal period of capital cost for gas-engine (GE) and micro gas-turbine (MGT) were 7.7 years and 9.1 years respectively. biogas utilization as Clean Natural Gas (CNG) shows lower capital cost and higher profit than that of CHP system. CNG gasificaion after biogas purification is likely the best alternative for Biogas utilization which have more economic potential than CHP system. The withdrawal period of capital cost appeared to be 2.3 years.

Purification and Characterization of Serine Protease Inhibitors from Dolichos lablab Seeds; Prevention Effects on Pseudomonal Elastase-Induced Septic Hypotension

  • Koo, Sun-Hyang;Choi, Yun-Lim;Choi, Su-Kyung;Shin, Young-Hee;Kim, Byeong-Gee;Lee, Bok-Luel
    • BMB Reports
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    • v.33 no.2
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    • pp.112-119
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    • 2000
  • Three kinds of serine protease inhibitors, members of the Bowman-Birk trypsin inhibitor, were purified from Dolichos lablab seeds and named Dolichos protease inhibitor 1, 2 and 3 (DI-1, DI-2 and DI-3), respectively. Each inhibitor showed a single band with gel mobility at around 15.9, 12.1 and 14.6 kDa on 20% SDS-PAGE under reducing conditions. To characterize inhibitory specificity, the inhibition constant (Ki) for these inhibitors was measured against several known serine proteases. All three Dolichos protease inhibitors (DI-1, DI-2 and DI-3) inhibited the activity of trypsin and plasmin, but had no effect on thrombin and kallikrein (either for human plasma kallikrein or for porcine pancreas kallikrein). DI-1 inhibited chymotrypsin most effectively (Ki = $3.6{\times}10^{-9}\;M$), while DI-2 displayed inhibitory activity for porcine pancreatic elastase (Ki = $6.2{\times}10^{-8}\;M$). Pre-treatment of the 33 mg/kg of DI-mixture (active fractions from $C_{18}$ open column chromatography that included DI-1, DI-2 and DI-3) inhibited the induction of pseudomonal elastase-induced septic hypotension and prevented an increase in bradykinin generation in pseudomonal elastase-treated guinea pig plasma. Also, the increase of kallikrein activity, by injection of pseudomonal elastase, was inhibited by the pretreatment of the DI-mixture in a guinea pig. Since the DI-mixture had no inhibitory effect on kallikrein activity when Z-Phe-Arg-MCA was used as a substrate in vitro, its inhibitory activity in the pseudomonal elastase-induced septic hypotension model might not be due to a direct inhibition of plasma kallikrein in the activation cascade of the Hageman factor and prekallikrein system. These results suggest that the Dolichos DI-mixture might be used as an inhibitor in pathogenic bacterial protease-induced septic shock.

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Purification of Antioxidant substance from the stem bark of Rhus verniciflua

  • Kim, Jung-Bae
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 2001.12a
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    • pp.126-126
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    • 2001
  • The Rhus verniciflua contains alkly(en)-catechol type allergens with a saiurated or unsaturated alkly chain of 15 or 17 carbon atoms. It has been recognized as an extremely active allergen causing skin reactions similar In poison ivy. The allergic contact dermatitis induced by the urushiol is known to be mediated be T lymphocytes whicht specifically recognize the hepten urushiol. Therefore. direct use of this plant as a medicinal purpose might imply a considerable hazard in Korea. In this study, using the established method for the detoxification from the stem bark of Rhus verniciflua, an strong antioxidant substance was isolated and characterized DPPH (diphenypricryl hydrazyl) assay measures hydrogen atom-donating activity and hence provides a measure of free radical scavenging antioxidant activity. DPPH, a purple-colored stable free radical, is reduced to yellow-colored diphenylpicryl hydrazine by antioxidants to deducing agents. Antioxidative effects of the water extract from RV were measured by DPPH assay. Twenty microliters of the extract was added to 1ml of 100mM DPPH solution in ethanol The mixture was shaken and left to stand for 10min at room temperature. The crude water extracts was purified by using HPLC method with a DEAE (anionic type), CN, ODS column. The purified compound remained stable at pH 3.0-6,0, but unstable above pH 6.5. It was stable heat at 10$0^{\circ}C$ for 4 hours, but still had about 80% of residual activity after treatment at 10$0^{\circ}C$ for 5 hours. The elemental composition of the HR-EI mass spectrum at m/z 170.02 was estimated the empirical formula as $C_{7}$ $H_{6}$ $O_{5}$. $C_{10}$ $H_4$ $O_2$N$_1$, $C_{5}$ $H_4$ $O_4$N$_3$, $C_{8}$$H_2O$$_1$N$_4$. In antimicrobial test, no inhibition was observed against Gram-positive and negative bacteria. This compound was stronger than that of commercial antioxidant by DPPH test, such as BHT, BHC at the same concentration (20$\mu$g/ml).ml).

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Analysis of Field Experimental Data for Water Quality Improvement of Tributary Stream to Estuarine Reservoir Using Constructed Wetland System (인공습지를 이용한 하구담수호 유입하천수 수질개선 현장실험결과 분석)

  • Ham, Jong-Hwa;Yoon, Chun-G.;Koo, Won-Seok;Kim, Hyung-Chul;Shin, Hyun-Bhum
    • Journal of The Korean Society of Agricultural Engineers
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    • v.46 no.5
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    • pp.141-153
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    • 2004
  • Wetland system is widely accepted as one of natural water purification systems around the world for nonpoint sources pollution control. Constructed wetlands have become a popular technology for treating contaminated surface and waste water. In this study, the field experiment to reduce nonpoint source pollution loadings from polluted stream waters using wetland system was performed from June 2002 to March 2004. Four wetlands were used and the size of each one was 0.8ha. Water of Dangjin stream flowing into Seokmun estuarine reservoir was pumped into wetlands. Inflow and hydraulic residence time of the system was 500 $m^3$/day∼1,500 $m^3$/day, 2∼5 days, respectively. After 2 year operation, plant-coverage of the wetlauds was about 70% from bare soil surface at initial stage . Average water quality of the influent was $BOD_5$ 4.17 mg/L, TSS 18.45 mg/L, T-N 4.32 mg/L, and T-P 0.30 mg/L. The average removal rate of $BOD_5$, TSS, T-N and T-P during the study period was 5.6%, 46.6%, 45.7%, and 54.8%, respectively. Organic ($BOD_5$) removal rate was low and the reason might be low influent concentration. Wetland removal rate of T-P was about 10% higher than T-N. Performance of the experimental system was compared with existing data base (NADB), and it was within the range of general system performance. Overall, the wetland system was found to be adequate for treating polluted water stream with stable removal efficiency even during the winter period. Most of the nonpoint source pollutions from watershed are transported by streams or ditches, and they could be controled by constructed wetland system before entering the lake or reservoir.

Evaluation of Ecological Function of Mangrove Soil on Absorbing Heavy Metals: A Case Study from the Dongzhaigang Mangrove in China

  • Xin, Kun;Huang, Xing;Zhou, Qingqing;Chen, Zhili
    • Journal of Ecology and Environment
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    • v.33 no.1
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    • pp.15-18
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    • 2010
  • Mangroves are special plant communities that live along intertidal zones in tropical and subtropical areas. They are regarded as one of the most important types of natural ecosystem in the world because of the many ecosystem functions that they perform, of which water purification is the most complex. Mangrove ecosystems are conducive to the deposition and retention of heavy metals. So it is important to understand the impact of heavy metals on mangrove ecosystems, and especially on soil subsystems. We examined the levels of heavy metals in the soil of mangroves in the Dongzhaigang Mangrove National Nature Reserve. Dongzhaigang, the first mangrove nature reserve established in China, is located south of Haikou in Hainan Island and encompasses $33.37\;km^2$, of which mangroves comprise $20.56\;km^2$. To assess the impact of human activities, we collected a large number of soil samples in four sampling areas (the protection station, the harbor, a tour area, and Yeboluo island) in the study area. We measured the concentrations of Cu, Pb, Zn and Cd in the soil samples using the spectra of polyatomic molecules. The average concentrations of Cu, Pb, Zn and Cd were $5.04\;{\mu}g/g$, $10.36\;{\mu}g/g$, $20.06\;{\mu}g/g$ and $0.06\;{\mu}g/g$, respectively, and the heavy metal concentrations were lowest in the protected area, highest in the harbor, and intermediate in Yeboluo Island and the tour area. The heavy metal concentrations in the soil collected from different sample plots are related not only to the physical and chemical properties of the soil, but also to the heavy metal emitted by nearby pollution sources. Our analysis indicates that tourist boats are the main pollution sources in the study area.

A Study on Purification of Water Works by Multi-filter Bed Method with Aeration(Appliction of Biological Oxidation by Aerobic Microorganism) (상수 정화법에 관한 연구 (폭기식 다단여과에 의한 호기성균의 생물화학적 산화작용을 응용))

  • Chung, Yo-Han;Moon, Jae-Kyu;Jhoo, Heung-Kyu;Seo, Hwa-Jung
    • Journal of Preventive Medicine and Public Health
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    • v.5 no.1
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    • pp.43-48
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    • 1972
  • 5 bed sand filter, applying biological oxidation, was designed and studied on the treatment of water works. Never using any coagulant agent (drugs), which may cause water pollution in pre-treatment of head water, the auther attempt a high rate filtration by the microorganism (nitrofication bacteria) end plant which populate in multi layer sand beds. The result are as follows : In order to evaluate the oxygen effect on filtration, oxygen was injected in aeration tank attached to each filter tank while filtration, and $NH_3$ was tested as a representaiive ingredient. It was found out that the aeration method was more effective, with over 33% of $NH_3$ removal capacity, than the anerobic and this 5 bed filter showed double removal capacity of $NH_3$ by comparing with conventional sand bed (2 stage bed). According to the examination of two kind of head water, pre-treated with coagulant agent and activated carton, the filtration capacity was affected by the polluted condition of head water, resulting that lower value of pollution and slower velocity of filtration showed more efficiency of $NH_3$ removal. In this experiment $NH_3$ content tested in treated water had a fairly good correlation with others.

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Enzymatic Characteristics and Applications of Microbial Chitin Deacetylases (미생물 Chitin Deacetylase의 특성과 응용)

  • Kuk Ju-Hee;Jung Woo-Jin;Kim Kil-Yong;Park Ro-Dong
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.9-15
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    • 2005
  • Chitin deacetylase (CDA; EC 3.5.1.41) catalyzes the hydrolysis of N-acetamide bonds of chitin, converting it to chitosan. Chitosan has several applications in areas such as biomedicine, food ingredients, cosmetics, pharmaceuticals, and agriculture. In this paper, occurrence, assay and purification protocols, enzymatic characteristics, substrate specificity, and mode of action of microbial CDAs have been described. Several lines of evidence have substantiated the biological roles involved in cell wall formation and plant-pathogen interactions for fungal CDAs. The gene structure of CDAs has been compared with other family 4 carbohydrate esterases which deacetylate a wide variety of acetylated poly/oligo-saccharides. The use of CDAs for the conversion of chitin to chitosan, in contrast to the presently used chemical procedure, offers the possibility of a controlled, non-degradable process, resulting in the production of well-defined chitosan oligomers and polymers. Insect pathogen that can secrete high levels of chitin-metab­olizing enzymes including CDA can be a possible alternative for new pest management tools.