• Title/Summary/Keyword: Isolation & purification

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Isolation and Purification of Protoplasts from PorPhyra tenera Thalli (김엽체에서의 원형질체분리와 정제)

  • SONG Sang Ho;CHUNG Gyu Hwa
    • Journal of Aquaculture
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    • v.1 no.1
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    • pp.103-108
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    • 1988
  • Protoplasts were isolated enzymatically from thalli of Porphyra tenera in relation to its utilization in breeding. As the concentration of the enzymes and incubation period increased, so did the yield of protoplasts. When a 250 mg fresh weight of the thalli was incubated in the $12\%$ abalone digestive enzyme mixture at $22^{\circ}C$ for 3 hours, about $2.5\times10^6$ protoplasts were released. The size of protoplasts ranged between 9$\mu$m and 25$\mu$m, with an average of 14.5$\mu$m.

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Production and Purification of tazane Derivatives from the Plane Cell Cultures of Taxus Chinensis in Large-scale Process (식물세포 Taxus chinensis 의 대량 배양액으로부터의 Taxane 유도체 생산 및 정체)

  • 김진현
    • KSBB Journal
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    • v.15 no.4
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    • pp.398-401
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    • 2000
  • Taxiods inclusive paclitaxel were produced isolated and purified from plant cell cultures of Taxus chinensis in large-scale process. their structures were elucidated by spectroscopic analyses. These compounds were exactly identical as those in previous studies from the other biomasses of Taxus chinensis and also other species. Also the concentrations of these compounds were compared with the concentration of the paclitaxel in various batches of plant cell cultures. As paclitaxel concentration increased at the end of cell cultures. the concentrations of the other paclitaxel derivatives decreased. The profile of these taxoids production can provide information for better understanding of structure-activity relationships and biosynthesis Importantly it can be utilized as an useful parameter for the quality control of paclitaxel production.

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Antioxidant Activity of the Halophyte Ligustrum japonicum (염생식물 광나무(Ligustrum japonicum)의 항산화 활성)

  • Baek, Seung Oh;Kim, Hojun;Jeong, Heejeong;Ju, Eunsin;Kong, Chang-Suk;Seo, Youngwan
    • KSBB Journal
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    • v.30 no.6
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    • pp.275-282
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    • 2015
  • Dried samples of Ligustrum japonicum were extracted twice: with methylene chloride and with methanol (MeOH), respectively. The combined crude extracts were successively fractionated into n-hexane, 85% aqueous methanol (85% aq.MeOH), n-butanol (n-BuOH), and water fractions by liquid-liquid partition. Antioxidant activities of crude extract and its solvent fractions were evaluated by measuring authentic $ONOO^-$, and $ONOO^-$ generated from 3-morpholinsydnonimine (SIN-1) as well as degree of occurrence of intracellular ROS in HT 1080 cells, and genomic DNA oxidation. The 85% aq.MeOH and n-BuOH fractions exhibited the good antioxidant activity. Further purification of the 85% aq.MeOH fracition led to the isolation of Oleanolic acid (1), Maslinic acid (2), and Ursolic acid (3). All compounds showed the significant antioxidant effects in all assay systems.

Isolation of Angiotensin Converting Enzyme Inhibitory Component from the Seeds of Xanthium strumarium (창이자(蒼耳子)로부터 안지오텐신 전환효소 억제 유효 성분의 분리)

  • Lee, Yun-Mi;Kang, Dae-Gill;Kim, Myung-Gyu;Jang, Ji-Yeon;Lee, Ho-Sub
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.1
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    • pp.119-123
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    • 2005
  • In the courses of in vitro screening for the angiotensin converting enzyme (ACE) inhibitory activity of the various extracts from medicinal plants, n-BuOH soluble extract of the seeds of Xanthium strumarium was found to exhibit distinctive angiotensin converting enzyme (ACE) inhibitory activity. Bioassay-guided fractionation and purification of the n-BuOH soluble extract of the seeds of Xanthium strumarium afforded a new $xanthiazone-11-{\beta}-glucopyranoside$. The ACE activity was significantly inhibited by the addition of a new $xanthiazone-11-{\beta}-glucopyranosidein$ a dose-dependent manner of which $IC_{50}$ value was $21.8\;{\mu}g/ml$.

Effective Application of CF11 Cellulose for Detection of Apple scar skin viroid in Apple

  • Chung, Bong-Nam;Cho, In-Sook;Cho, Jeom-Deog
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.291-293
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    • 2009
  • The low virus titer in woody plant tissues and the presence of inhibitor compounds such as polyphenols, tannins and polysaccharides are common difficulties that compromise purification of plant viroids from their woody hosts. A simple, reliable method of RNA isolation using CF11 cellulose column on a microcentrifuge tube scale for detecting Apple scar skin viroid (ASSVd) in apple was developed. Total RNA extracted from leaf, woody bark and the fruit skin was used for reverse transcription. RT-PCR products could be detected from RNA prepared from dormant woody bark, fruit skin and fresh leaves with both the CF11 cellulose column method and NucliSens extractor in February, August and November. Meanwhile, with the RNeasy kit RT-PCR, products were detected only in leaves and not from bark or fruit skin. The PCR product, about 330 base pairs, was analyzed by agarose gel electrophoresis. The CF11 cellulose column method was effective for detecting ASSVd. The method enabled the processing of a large numbers of samples of dormant woody bark, leaf and fruit skin of apple.

Antioxidant Activity of Flavonoids Isolated from Vitex rotundifolia (순비기나무(Vitex rotundifolia)로부터 분리한 플라보노이드 성분의 항산화 활성)

  • Kim, You-Ah;Lee, Jung-Im;Hong, Joo-Wan;Jung, Myoung-Eun;Seo, Young-Wan
    • Ocean and Polar Research
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    • v.33 no.3
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    • pp.255-263
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    • 2011
  • The aim of this investigation was to evaluate antioxidant activity of crude extracts from the halophyte Vitex rotundifolia, their solvent fractions, and isolated compounds (1-3). Antioxidant capacity was determined by measuring DPPH radical, and authentic $ONOO^-$ and $ONOO^-$ generated from 3- morpholinsydnonimine (SIN-1) in vitro as well as degree of occurrence of intracellular ROS, NO and GSH in mouse macrophage Raw 264.7 cells. From comparative analysis, MeOH extract, n-BuOH, and 85% aq. MeOH solvent fractions showed significant antioxidant effect in DPPH radical and $ONOO^-$ assay systems. Activity-guided purification of n-BuOH and 85% aq. MeOH fractions led to the isolation of flavonoids 1-3. Among them, compound 1 exhibited excellent antioxidant effect in all bioassay systems tested. On the other hand, compounds 2 and 3 revealed potent inhibitory effect against $ONOO^-$ generated from SIN-1, comparable with the positive control penicillamine.

Isolation, Optimization, and Partial Purification of Amylase from Chrysosporium asperatum by Submerged Fermentation

  • Sanghvi, Gaurav V.;Koyani, Rina D.;Rajput, Kishore S.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.470-476
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    • 2011
  • A potent fungus for amylase production, Chrysosporium asperatum, was isolated from among 30 different cultures obtained from wood samples collected in the Junagadh forest, India. All of the isolated cultures were screened for their ability to produce amylase by submerged fermentation. Among the selected cultures, C. asperatum (Class Euascomycetes; Onygenales; Onygenaceae) gave maximum amylase production. In all of the different media tested, potato starch was found to be a good substrate for production of amylase enzyme at $30^{\circ}C$ and pH 5.0. Production of enzyme reached the maximum when a combination of starch and 2% xylose, and organic nitrogen (1% yeast extract) and ammonium sulfate were used as carbon and nitrogen sources, respectively. There was no significant effect of metal ions on enzyme activity. The enzyme was relatively stable at $30^{\circ}C$ for 20 min, and no inhibitory effect of $Ca^{+2}$ ions on amylase production was observed.

Topical Anti-inflammatory Activity of Dianemycin Isolated from Streptomyces sp. MT 2705-4

  • Lee, Song-Jin;Kim, Hyun-Pyo;Park, Byung-Keun;Ahn, Soon-Cheol;Lee, Hyun-Sun;Ahn, Jong-Seog
    • Archives of Pharmacal Research
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    • v.20 no.4
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    • pp.372-374
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    • 1997
  • In order to develop new anti-inflammatory agents having different action mechanisms compared with nonsteroidal and steroidal anti-inflammatory drugs, the culture broths of various actinomycetes isolated from soil were screened using an in vivo mouse ear edma assay and one strain (Streptomyces sp. MT 2705-4: KCTC 8651 P) was selected. Activity-guided purification led to the isolation of a polyether compound, dianemycin. Topically, dianemycin showed a potent anti-inflammatory activity in mouse ear edema induced by croton-oil or arachidonic acid.$ED_{50}$value of dianemycin was found to be 0.8 mg,/ear compared to 0.4 mg/ear of prednisolone in croton-oil ear edema. However, dianemycin did not show the inhibitory activity in UV-erythema and delayed hypersensitivity reaction. These results indicate that dianemycin is a potential topical anti-inflammatory agent.

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Isolation, Purification and Characterization of Polysaccharides that induce in vitro Immuno-Stimulation of Macrophases derived from Liquid Culture of Cordyceps militaris

  • Kwon, Jeong-Seok;An, Hyo-Sil;Hong, Eock-Kee
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.287-292
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    • 2005
  • The crude polysaccharides(C-CPF, C-CPM, C-CPB) derived from fruiting body, mycelia and mycelia free broth of cordyceps militaris were obtained by ethanol precipitation of hot water extracts. After a batch fermentation of C. militaris was carried out in a 5 L jar vessel, endo-polysaccharide and exo-polysaccharide were obtained. They were demonstrated as the hetero polysaccharides which were composed of glucsose, galactose and mannose by performed with HPAEC(high pH anion exchange chromatography) and conformation of random coil by its complex forming ability with congo red reagent. They were purified by ion exchange (DEAE-cellulose) and gel filtration chromatography. They were monitered by phenol-sulfuric acid method and Bradford method. The NO induction activities of crude polysaccharides and purified polysaccharides derived from mycelia free broth were enhanced rather than LPS(lipo polysaccharide) which was used as a general NO inducer. These effects presumably contibute to the antitumor activities. The homogenieties and molecular weights of polysaccharides were determined by using Sepharose CL-6B. The yield, molecular weights and NO induction activities of C-CPFN Fr.III, C-CPMN Fr.III, C-CPBN Fr.II were 0.387, 0.408 and 0.153, 127 K 210 K and 36 K, 40.79%, 88.72%, and 104.17%, respectively.

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Isolation and Purification of Several Substances Produced by Fusarium graminearum and Their Antimicrobial Activities (Fusarium graminearum이 생산하는 몇가지 물질의 분리정제 및 항균 활성)

  • 김병섭;김건우;이종규;이인원;조광연
    • Korean Journal Plant Pathology
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    • v.11 no.2
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    • pp.158-164
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    • 1995
  • 토마토의 엽권에서 분리한 Fusariym graminearum이 분비하는 물질은 벼 도열병균(Pyricularia oryzae)의 여러 종의 식물병원 진균에 대한 항균활성을 나타내었으며, 이러한 활성물질을 PDA에서 본 균을 배양 한 후 chloroform으로 추출하여 분리정제 하였다. HPLC에 의하여 5종류의 활성 물질을 분획하였으며, 그중 1번(F402) 화합물을 벼 도열병균(P. oryzae)을 포함한 22개 식물 병원 진균에 대하여 살균 활성범위를 조사한 결과, 이 화합물은 50$\mu\textrm{g}$/ml 농도에서 Pythium ultimum, Rhizoctonia solani, Sclerotinia sclerotiorum은 전혀 억제하지 못하였으며, Phytophthora spp., Cladosporium fulvum, Fusarium spp., Corynespora cassicola에는 어느 정도의 활성이 있었지만 낮게 나타났고, P. oryzae, Cochliobolus miyabeanus, Alternaria solani는 100% 억제하여 활성이 높게 나타났다. 또한 장내 세균에 대한 활성을 MIC로 비교할 때 Streptococcus pyogenes, Streptococcus faecium에 대하여는 각각 12.5, 25 $\mu\textrm{g}$/ml였고 Staphylococcus aureus는 25-50$\mu\textrm{g}$/ml으로 나타났으며, Pseudomonas aeruginosa, Salmonella typhimurium, Klebsiella aerogenes, Enterobacter cloacae에서는 100$\mu\textrm{g}$/ml 이상으로 활성이 나타나지 않았다. F402를 200$\mu\textrm{g}$/ml의 농도로 직접 살포한 식물체에서의 방제효과는 벼도열병, 벼 깨씨무늬병, 보리 흰 가루병에 대하여는 80%이상이었으나, 벼 잎집무늬마름병, 오이 잿빛곰팡이병, 토마토 역병, 밀 녹병에서는 낮았다.

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