• Title/Summary/Keyword: column chromatography

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Isolation of 3,4-Dihydroxybenzoic Acid with Antimicrobial Activity from Bark of Aralia elata (두릅수피에서 항미생물활성을 갖는 3,4- dihydroxybenzoic acid의 분리)

  • Ma, Seung-Jin;Ko, Byoung-Seub;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.807-812
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    • 1995
  • The methanol extracts of Aralia elata bark showed antimicrobial activities against bacteria, yeast, fungi. The solvent fractionated acidic fraction that showed the activity was then successively purified with silica gel adsorption column chromatography, Sephadex LH-20 column chromatography, silica gel partition column chromatography, HPLC and TLC. The isolated major active substance was identified as 3,4-dihydroxybenzoic acid by MS, GC-MS, IR, $^{1}H-NMR\;and\;^{13}C-NMR$. The content of 3,4-dihydroxybenzoic acid was 0.869㎎/g in dried bark of Aralia elata.

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Biochemical Characteristics of Lectins Isolated from Lentinula edodes (표고버섯으로부터 분리한 렉틴의 생화학적 특성)

  • Kim, Young-Sin;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • v.29 no.4
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    • pp.79-88
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    • 2001
  • Lectin was isolated from shiitake mushroom (Lentinula edodes) with 0.15 M NaCl solution, and purified by the following procedures : precipitation by ammonium sulfate, anion exchange column chromatography on DEAE Sephadex A-50 and hydroxyapatite column chromatography. The fresh pileus part of the mushroom contained more than two times of lectin compared to the stipe part, and lectins and its activity were reduced by heating. The extraction yield of crude lectin was 46.03%, 28% yield after purification on on DEAE Sephadex A-50 column chromatography. Some amino acids, aspartic acid, serine, alanine and histidine, were increased by purification process. Relatively low molecular weight parts of lectin had the agglutinating activity for rabbit blood, and its molecular weight was about 23 kDa The molecular weights of purified lectins, LA-a and LB-b, by the hydroxyapatite column chromatography were 24 kDa and 23 kDa, respectively.

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Separation and Identification of Antimicrobial Substances from Prunus mume extract (매실추출물로부터 항균물질의 분리 및 구조동정)

  • Park, Woo-Po;Lee, Seung-Cheol;Kim, Sung-Yong;Choi, Sung-Gil;Heo, Ho-Jin;Cho, Sung-Hwan
    • Food Science and Preservation
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    • v.15 no.6
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    • pp.878-883
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    • 2008
  • Prunus mume was extracted using various solvents including ethyl ether, ethyl acetate and n-butanol. The extracts were fractionated by column chromatography. Antimicrobial compounds (A, B and C) in fractions showing antimicrobial activity were purified by Sephadex LH 20 column chromatography, and identified by $^{1}H$- and $^{13}C$-NMR analysis as isoeugenol, nomilin and $\beta$-sitosterol, respectively.

Purification and Characteristics of Tyrosinase Inhibitor Produced by Actinomycetes F-97 (방선균 F-97이 생산하는 Tyrosinase 저해제의 정제 및 특징)

  • Bang, Byung-Ho;Rhee, Moon-Soo;Kim, Jin-O;Yi, Dong-Heui
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.153-158
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    • 2008
  • An actinomycetes F-97 producing tyrosinase inhibitor was isolated from soil samples. Isolation and purification of tyrosinase inhibitor produced by F-97 was performed as follows: IRC-120 ($NH_4^+$ type) column chromatography, silica gel column chromatography, $C_{18}$ column chromatography and Sephadex LH-20 column chromatography were used successively after the centrifuged supernatant was adjusted to pH 4.0. To identify the purity of the inhibitor, octadecylsilyl(ODS) HPLC was carried out with 5% methanol as a mobile phase. Finally, the purification yield of a tyrosinase inhibitor was 5.24%. The inhibitor was very soluble in water, methanol and ethanol but insoluble in acetone, butanol, ethylacetate and chloroform. The ${\lambda}_{max}$ value of this inhibitor in water was 194nm under UV light. The biochemical test of the inhibitor was positive in Molish, Benedict, cone. $H_2SO_4$, and $KMnO_4$ tests but negative in iodine, ninhydrin, Million, Sakaguchi, xanthoproteic and Emerson tests. The tyrosinase inhibitor was stable against heat treatment of $100^{\circ}C$ for 50 minutes and pH $4{\sim}9$. The $IC_{50}$ value of this inhibitor was $19.2{\mu}g/ml$ for mushroom tyrosinase. In $1,000{\mu}g/ml$ inhibitor concentration, inhibition zone was 27 mm for Streptomyces bikiniensis NRRL B-1049. The inhibition of F-97 against mushroom tyrosinase was competitive with tyrosine.

Development of Miniaturized Automatic Chromatography System for Column Performance Validation : MiniValChrom

  • Park, Jae-Ha;Lee, Eun-Gyu
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.147-150
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    • 2000
  • Bioprocess chromatography is probably the most widely used unit operation in biopharmaceutics manufacturing. To obtain a good quality product reproducibly, the chromatography process validation should be performed carefully. We developed a miniaturized automatic chromatography system, MiniValChrom, for column performance validation.

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Production, Purification and Antifungal Activity of Antibiotic Substances Produced by Pseudomonas aeruginosa Strain B5

  • Kim, Beom-Seok
    • Journal of Microbiology and Biotechnology
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    • v.3 no.1
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    • pp.12-18
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    • 1993
  • Pseudomonas aeruginosa strain B5 with antagonistic activity against Phytophthora capsici and Magnaporthe grisea, was isolated from pepper-growing soil. From the culture of P. aeruginosa strain B5 grown on King's medium B, antibiotic substances were purified using XAD-2 column chromatography. XAD-2 eluates inhibited not only the mycelial growth of P. capsid and M. grisea, but also the development of Phytophthora blight on pepper plants. The crude antibiotic substances were further purified by using silica gel column chromatography, Sephadex LH-20 column chromatography, thin layer chromatography on silica gel plates, and high performance liquid chromatography. Silica gel column chromatogrphy gave good separation of the four antibiotic substances. The pure antibiotics P1, P2, and P3 finally purified by preparative HPLC inhibited the mycelial growth of P. capsici, at concentrations from 7 to 10 $\mu g/ml$. Only P1 and P2 had antifungal activity against M. grisea at 8 $\mu g/ml$. P1 and P3 were highly inhibitory to the mycelial growth of Botryosphaeria dothidea and Botrytis cinerea at relatively low concentrations. However, the three antibiotics had no antifungal activity against Rhizoctonia solani. The chemical structures of these antibiotics are being identified.

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Isolation and Characterization of Cinnamic Acid with Antimicrobial Activity from Needle of Pinus densiflora (솔잎에서 항미생물 활성을 갖는 Cinnamic Acid 의 분리 및 동정)

  • Kuk, Ju-Hee;Ma, Seung-Jin;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.29 no.4
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    • pp.823-826
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    • 1997
  • The EtOAc extracts from needles of Pinus densiflora showed antimicrobial activities against bacteria, yeast and fungi. The antimicrobial principle was successively purified by solvent fractionation, silica gel adsorption column chromatography and Sephadex LH-20 column chromatography. The active substance was further purified by HPLC using $C_{18}$ column. The active substance was identified as trans-cinnamic acid by MS, $^{1}H-NMR\;and\;^{13}C-NMR$. The amount of cinnamic acid was $9.27\;{\mu}g$ Per gram of fresh needle of Pinus densiflora.

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Purification and Characterization of Extracellular Protease form Psychrotrophic Antarctic Bacteria (남극에서 분리한 저온성 세균 유래 단백질 분해 효소)

  • 조기웅;방지헌;홍혜원;박승일;이윤호
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.254-259
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    • 2002
  • A psychrotrophic bacterium was isolated from Antarctic marine sediment and identified as Shewanella sp. species based on the biochemical properties and 16S rRNA sequence, and designated as Shewanella sp. L93. Extracellular protease produced by this strain was purified through ammonium sulfate precipitation, High-Q column chromatography, first gel permeation chromatography, BioScale Q2 ion exchange chromatography and second gel permeation chromatography, and basic properties of this enzyme were investigated.

Study on the Development of Simultaneous Analytical Method for the Residual Organic Chloride Pesticides by Gas Chromatography (기체 크로마토그래피를 이용한 유기 염소계 잔류 농약 동시 분석 방법 개발에 관한 연구)

  • Kim, Woo-Seong;Lee, Bong-Hun;Park, Heung-Jai
    • Journal of Environmental Science International
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    • v.5 no.5
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    • pp.561-567
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    • 1996
  • A method for the simultaneous analysis of 31 residual organic chloride pesticides was studied using gas chromatography. Prepared analytical samples were injected to gas chromatography (HP 5890 Series II plus) on the Ultra-2 column with ECD. The packing materials for column were changed as the following reagents ; florisil and alumina N, The residual solution was loaded to column and was elected pith erection solvents ; ether : benzene (2 : 8) solution, hexane : benzene (1 : 1) solution, dichloromethane, acetone, and methanol. The analytical results showed that 6 kinds of organic chlorides were not detected when florisil (first condition) was used as the column packing material. The nondetected 6 kinds of organic chlorides in the first analytical condition were detected and the recoveries of thrin-pesticides were increased, in particular, captan and captafol, but the recoveries of benzene hexachloride compounds were decreased when dichloromethane and methanol were added as elution solvents (pac'king material was florisil as in the first condition). The recoveries of dichlornuanid, chlorofenvinfos, folpet, and dicofol were increased and that of aldrin was increased, but those of captan and captafol were not good when alumina N was used as the packing material. To detect simultaneously thrin-pesticides, captan, and captafol, florisil and alumina N were used as the packing materials. The elution result showed that captan and captafol were not detected. This was because the column was activated insufficiently. The analytical method was the best (31 kinds of organic chlorides in the residual pesticides were detected sharply and showed high sensitivity) when the column (packing materials were florisil and alumina N: together) was fuliy activated and the impurities were removed using various elution solvents.

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