• Title/Summary/Keyword: Natural purification

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Purification and Characterization of Xylanase II from Trichoderma koningii ATCC 26113 (Trichoderma koningii ATCC 26113으로부터 Xylanase II의 순수분리 및 특성)

  • Kim, Hyun-Ju;Kang. Sa Ouk;Hah, Yung-Chil
    • Korean Journal of Microbiology
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    • v.31 no.2
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    • pp.157-165
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    • 1993
  • A 1, 4-.betha.-D-xylanase, designated as xylanase II, was purified from the culture filtrate of Trichoderma koningii ATCC 251131 by column chromatography on Sephadex G-75, SP-Sephadex C-50, DEAE-Sephadex A-50 and Sephadex G-50 with an overall yield of 6.97%. It has a molecular weight of 21.000 and an isoelectric point of 9.4. The enzyme activity is optimal at pH 5.0 and at a temperature of 50.deg.C. Xylanase II is stable up to 50.deg.C, while 40 and 90% of its activity are lost after the incubation for 30 and 60 min at 60.deg.C. The enzyme degrades xylan with relatively high activity, as well as carboxymethylcellulose and Avicel. Its $K_{m}$ values for oat-spelt xylan, larchwood xylan and Avicel are 7.48, 1.98 and 13.33 mg/ml, respectively. The hydrolysis products of oat-spelt xylan by xylanase II are xylose, xylobiose, xylotriose and arabinoxylotriose, while the reaction products of larchwood xylan are xylose, xylobiose, xylotriose and small amount of higher oligomers. The action paterns of the enzyme demonstrate that xylanase II is endo-enzyme.

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In vitro Regeneration of Phragmites australis through Embryogenic Cultures

  • Lee Jeong-Sun;Kim Chang-Kyun;Kim In-Sung;Lee Eun-Ju;Choi Hong-Keun
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.21-25
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    • 2006
  • Phragmites australis (reed) has received much attention as being one of the principle emergent aquatic plants for treating industrial and civil wastewater. Plant regeneration via plant tissue culture in p. australis was investigated. Three types of callus were identified from seeds on N6 medium plus 4.5 UM 2,4-dichlorophenoxyacetic acid (2,4-D). Yellow compact type showed the best redifferentiation, whereas white compact type and yellow friable were not competent to differentiate into plane. Solid medium culture was better than liquid suspension culture for enhancing callus growth when N6 medium supplemented with 4.5 ${\mu}M$ 2,4-D was used. Phytagel, as a gelling agent, was superior to agar in plant regeneration on N6 medium, supplemented with 9.4 ${\mu}M$ kinetin and 0.54 ${\mu}M$ $\alpha$-naphthaleneacetic acid (NAA). Transfer of the plantlets regenerated from kinetin and NAA-supplemented N6 medium to growth regulator-free MS medium enhanced the further development of the plantlets. Plantlets on subsequently grown to maturity when tansferred to potting soil. The regenerated plants exhibited morphologically normal. The system for plant regeneration of P. australis enables to propagate elite lines on a large scale for water purification in the ecosystem

Purification and Charaterization of Antifungal Chitinase from Indigenous Antagonistic Microorganism Serratia sp. 3095

  • Lee, Eun-Tag;Kim, Sang-Dal
    • Journal of Applied Biological Chemistry
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    • v.42 no.1
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    • pp.7-11
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    • 1999
  • An extracellular chitinase of the selected strong antifungal microorganism, Serratia sp. 3095, was purified by salting out, affinity adsorption, Sepadex G-100 gel fitration, Sepadex G-75 gel fitration and DEAE Sepadex A-50 chromatography. The molecular weight of the purified chitinase was estimated to be 62,000 dalton by SDS-PAGE. Optimal pH and temperature of the chitinase were pH 7.5 and 45, respectively. The enzyme retained more than 80% of the activity between pH 5.5 and pH 10.5, and below $50^{\circ}C$ but was unstable above $60^{\circ}C$, below pH 5.0. The activity of the chitinase was inhibited about 60% by $Sn^{2+}$, 40% by $Hg^{2+}$ and $Ag^+$, 70% by AHA, 40% by iodoacetate, 35% by thiourea and p-CMB, but stabilized by SDS. $K_m$ value of the purified chitinase was 3.68 mg/ml for colloidal chitin. The chitinase from Serratia sp. 3095 showed antifungal activity to Fusariurm solani.

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Inhibitory Activity of Chinese Medicinal Plants on Nitric Oxide Synthesis in Lipopolysaccharide -Activated Macrophages

  • Ryu, Jae-Ha;Ahn, Han-Na;Lee, Hwa-Jin;Feng, Li;Qun, Wen-He;Han, Yong-Nam;Han, Byung-Hoon
    • Biomolecules & Therapeutics
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    • v.9 no.3
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    • pp.183-187
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    • 2001
  • Nitric oxide (NO) produced in large amounts by the inducible nitric oxide synthase (iNOS) is known to be responsible for the vasodilation and hypotension observed in septic shock and inflammation. The inhibitors of iNOS, thus, may be useful candidate for the treatment of inflammatory diseases accompanied by the overproduction of NO. We prepared alcoholic extracts of Chinese medicinal plants and screened their inhibitory activity against NO production in lipopolysaccharide (LPS)-activated macrophages. Among the 80 kinds of extracts of herbal drugs, 15 extracts showed potent inhibitory activity of NO production above 80% at the concentration o$50\mu\textrm{g}/ml$. These potent extracts showed dose dependent inhibition of NO production of LPS-activated macrophages at the concentration of 50, 30,$10\mu\textrm{g}/ml$. Especially, Rhus chinensis, Senecio scandens and Wikstroemia indica showed most potent inhibition above 50% at the concentration of $10\mu\textrm{g}/ml$. These plants are promising candidates for the study of the activity-guided purification of active compounds and would be useful for the treatment of inflammatory diseases and endotoxemia accompanying the overproduction of NO.

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A Large Scale Separation & Purification of Cyanobacterial Toxins (남조류 독성물질의 대량분리 및 정제)

  • Yoon, Suk Chang;Park, Keun Young;Pyo, Dong Jin
    • Journal of the Korean Chemical Society
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    • v.42 no.1
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    • pp.51-56
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    • 1998
  • It is very difficult to separate and purify the microcystins, cyanobacterial toxins since it exist in a trace level in natural lakes. In this paper, we developed a new analytical method to separate and purify the microcystin RR and LR from the freeze-dried cyanobacterial cells in natural lakes. We used 7.5 g silica gel as a stationary phase and ethyl acetate: isopropanol: water (30: 45: 25) as a mobile phase and microcystins were eluted using an open column. The eluting solvent was collected in a small bottle at the intervals of 3 mL and the fractions were chromatographed with HPLC to confirm the microcystin RR and LR.

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Feedback Control using Dual O2 Sensors for Improving the Conversion Efficiency of a Three-way Catalyst in a Heavy-duty CNG Engine (CNG 대형엔진에서 이중 O2 센서를 활용한 피드백 제어를 통한 삼원촉매 정화효율 향상)

  • Yoon, Sungjun;Lee, Junsun;Park, Hyunwook;Lee, Yonggyu;Kim, Changup;Oh, Seungmook
    • Journal of ILASS-Korea
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    • v.24 no.4
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    • pp.163-170
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    • 2019
  • In this study, feedback logic using dual O2 sensor values were developed to increase the purification capability of a three-way catalyst (TWC) in a compressed natural gas (CNG) engine. A heavy-duty inline 6-cylinder engine was used and the CNG was supplied to the engine through a mixer. This study consists of two main parts, namely, the proportional integral (PI) control with a front O2 sensor and the feedback control with dual O2 sensors. In the PI control experiment, effects of various parameters, such as P gain, I gain, and lean delay, on the TWC capability were identified. Based on the results of the PI control experiment, the feedback logic using dual O2 sensor values were developed. In both cases, the nitrogen oxides (NOX) emissions were nearly zero. However, the carbon monoxide (CO) emissions were reduced significant in the feedback logic with dual O2 sensors than in the PI control with the front O2 sensor.

Purification and Properties of HPS (Halitosis Prevention Substance) Isolated from Cumin (Cuminum cyminum L.) Seed

  • Kang, Eun-Ju;Ryu, Il-Hwan;Lee, Kap-Sang
    • Food Science and Biotechnology
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    • v.14 no.5
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    • pp.621-627
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    • 2005
  • Halitosis is mainly caused by the presence of volatile sulfur-containing compounds (VSC's) produced by proteolytic periodontopathic bacteria in the oral cavity. Various mouth-rinses have been offered on the market as solutions to reduce halitosis. The aim of this study was to find a potent substance for the prevention of halitosis. The halitosis prevention substance (HPS) from cumin seed powder was purified by solvent extraction, silica gel column chromatography and preparative TLC to yield an oil phase (0.98%). Instrumental analysis such as FT-IR, $^1H$-NMR and $^{13}C$-NMR showed that HPS contained an -OH group, -HC=CH-, -COO-, and long chain acyl group. HPS was therefore determined to be 2-hydroxyethyl-${\beta}$-undecenate. HPS inhibited the growth of Fusobacterium nucleatum and Porphyromonas gingivalis, by 72.44% and 64.37% at $1{\times}10^{-2}\;M$, and by 99.85% and 91.62% at $5\;{\times}\;10^{-2}\;M$, respectively. It also inhibited the activity of L-methionine-${\alpha}$-deamino-${\gamma}$-mercaptomethane-lyase (METase), which was produced by oral microbes. Furthermore, the VSC production by oral microbes in the human mouth air decreased with increasing HPS concentration. These results suggested that HPS from cumin seed is an efficient halitosis prevention agent.

Separation and Purification of Antihypertensive Substances from Edible Seaweeds (해조류로부터 항고혈압성분의 분리정제)

  • Lee, Seung-Joo;Ha, Wang-Hyun;Choi, Hye-Jin;Cho, Soon-Yeong;Choi, Jong-Won
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.5
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    • pp.421-427
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    • 2010
  • To isolate natural antihypertensive substances from edible seaweeds, we screened for and separated active compounds contained in natural Underia pinnatifida, cultural Underia pinnatifida, Laminaria japonica, Sporophylls and Agarum cribrosum. They were extracted using room temperature water, boiling water, acetone, and methanol in turn or using room temperature water, ether, acetone, methanol and boiling water in order. The in vitro antihypertensive activity was quantified as inhibitory efficacy against angiotensin-I converting enzyme (ACE), which is a factor inducing hypertension. For all of the seaweeds tested, the fractions soluble in room temperature water and in boiling water showed the strongest ACE inhibitory effect among the extracted fractions. Conversely, the methanol-extracted fractions for all of the seaweeds tested showed no antihypertensive activity. While the ether and acetone fractions had slight antihypertensive effects. The compounds in the aqueous extracts that had antihypertensive activity were presumed to be polysaccharides, such as fucoidan and alginate.

Synthesis and Characterization of Oligonucleotides Containing Site-Specific Bulky $N^2$-Aralkylated Guanines and $N^6$-Aralkylated Adenines

  • Moon, Ki-Young;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • v.23 no.2
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    • pp.139-146
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    • 2000
  • 7- Bromomethylbenz[a]anthracene is a known mutagen and carcinogen. The two major DNA adducts produced by this carcinogen, i.e., $N^2$-(benz[a]anthracen-7-yl methyl)-2'-deoxyguanosine (2, b[a]$a^2$G) and $N^6$-(benz[a]anthracen-7-ylmethyl)-2'-deoxyadenosine (4, b[a]$a^6$/A), as wel 1 as the simpler benzylated analogs,$N^2$-benzyl-2'deoxyguanosine (1, $bn^2$G) and $N^6$-benzyl-2'-deoxyadenosine (3, $bn^6$/A), were prepared by direct aralkylation of 2'-deoxyguanosine and 2'-deoxyadenosine. To determine the site-specific mutagenicity of these bulky exocyclic amino-substituted adducts, the suitably protected nucleosides were incorporated into 16-base oligodeoxyribonucleotides in place of a normal guanine or adenine residues which respectively are part of the ATG initiation codon for the lac Z' \alpha-complementation gene by using an in situ activation approach and automated phosphite triester synthetic methods. The base composition and the incorporation of the bulky adducts into synthetic oligonucleotides were characterized after purification of the modified oligonucleotides by enzymatic digestion and HPLC analysis.

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Biotransformation of Eugenol via Protocatechuic Acid by Thermophilic Geobacillus sp. AY 946034 Strain

  • Giedraityte, Grazina;Kalediene, Lilija
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.475-482
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    • 2014
  • The metabolic pathway of eugenol degradation by thermophilic Geobacillus sp. AY 946034 strain was analyzed based on the lack of data about eugenol degradation by thermophiles. TLC, GC-MS, and biotransformation with resting cells showed that eugenol was oxidized through coniferyl alcohol, and ferulic and vanillic acids to protocatechuic acid before the aromatic ring was cleaved. The cell-free extract of Geobacillus sp. AY 946034 strain grown on eugenol showed a high activity of eugenol hydroxylase, feruloyl-CoA synthetase, vanillate-O-demethylase, and protocatechuate 3,4-dioxygenase. The key enzyme, protocatechuate 3,4-dioxygenase, which plays a crucial role in the degradation of various aromatic compounds, was purified 135-fold to homogeneity with a 34% overall recovery from Geobacillus sp. AY 946034. The relative molecular mass of the native enzyme was about $450{\pm}10$ kDa and was composed of the non-identical subunits. The pH and temperature optima for enzyme activity were 8 and $60^{\circ}C$, respectively. The half-life of protocatechuate 3,4-dioxygenase at the optimum temperature was 50 min.