• Title, Summary, Keyword: chromatography

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Brassinosteroid Substances in Immature Zea mays Seeds (옥수수 종실의 Brassinosteroid 활성물질 탐색)

  • 박근형;김선재현규환
    • KSBB Journal
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    • v.8 no.3
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    • pp.300-305
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    • 1993
  • In order to explore the brassinosteroid-active components in Zea mays seeds, the methanol extract was purified by the sequences of solvent fractionation, silica gel adsorption chromatography, Sephadex LH-20 chromatography, charcoal adsorption chromatography and Bondesil 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. Brassinosteroid substances in separated active fractions were identified as castasterone and teasterone by HPLC. The content of brassinosteroid in Zea mays seeds as converted into brassinolide was 3-8ng/g fresh weight.

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Brassinosteroid substances in immature Perilla frutescense seeds (들깨의 brassinosteroid 활성물질)

  • Park, Keun-Hyung;Kim, Seon-Jae;Hyun, Kyu-Hwan
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.197-202
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    • 1993
  • In order to explore the brassinosteroid-active component in Perilla frutescense, methanol extract of immature seeds was purified by sequences of solvent fractionation, silica gel adsorption chromatography, Sephadex LH-20 chromatography, charcoal adsorption chromatography and Bondesil 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 seperated by silica gel adsorption chromatography. The seperated main and minor active brassinosteroid fractions were identified as castasterone and homobrassinolide, respectively, by HPLC. We acknowledge that our work is probably the first report of endogenous brassinosteroid in Perilla frutescense. The content of brassinosteroid in Perilla frutescense as converted into brassinolide was $0.5{\sim}0.8\;ng/g$ fresh weight.

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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.

Purifications of Phenoxyethanol Galactoside and Chlorphenesin Galactoside using Solvent Extraction followed by Gel Chromatography (Solvent Extraction과 Gel Chromatography를 이용한 Phenoxyethanol Galactoside와 Chlorphenesin Galactoside의 정제)

  • Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.4
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    • pp.954-961
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    • 2017
  • We investigated the purifications of PE-gal and CPN-gal, synthesized by transgalactosylation reaction using recombinant ${\beta}$-gal. The reaction mixture containing PE and PE-gal was first mixed with EA, and thereafter PE and PE-gal were distributed in two-phase (EA/water) system. In this system, PE and PE-gal was selectively moved into EA and water phase, respectively. Then, the water phase was collected, and silica gel chromatography was carried out using the collected water phase. Finally, we compared two purified PE-gal samples using HPLC and TLC analysis, in which the one was purified only by silica gel chromatography and the other was purified by EA extraction followed by silica gel chromatography. In the latter case, the residual PE was almost completely removed, whereas, in the former case, the residual PE was remained remarkably. Additionally, the purification yield of PE-gal was about 21% on the basis of mole. In the same purification protocol, CPN-gal was able to be purified using EA extraction followed by silica gel chromatography, in which the residual CPN was almost removed when CPN-gal was purified by EA extraction followed by silica gel chromatography.

Purification of Peptide Components including Melittin from Bee Venom using gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis (Gel filtration chromatography와 propionic acid/urea polyacrylamide gel electrophoresis를 이용한 봉독 성분의 분리)

  • Choi, Young-Chon;Choi, Suk-Ho;Kwon, Ki-Rok
    • Journal of Pharmacopuncture
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    • v.9 no.2
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    • pp.105-111
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    • 2006
  • Objectives : This study was conducted to carry out Purification of Melittin and other peptide components from Bee Venom using gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis Methods : Melittin and other peptide components were separated from bee venom by using gel filtration chromatography on Sephadex G-50 column in 0.05M ammonium acetate buffer. Results : Melittin and other peptide components were separated from bee venom by using gel filtration chromatography on Sephadex G-50 column in 0.05M ammonium acetate buffer. The fractions obtained from gel filtration chromatography was analyzed by using SDS-PAGE and propionic acid/urea polyacrylamide gel electrophoresis. The melittin obtained from the gel filtration contained residual amount of phospholipase $A_2$ and a protein with molecular weight of 6,000. The contaminating proteins were removed by the second gel filtration chromatography. Conclusion : Gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis are useful to separate peptide components including melittin from bee venom.

A study on the Detection of Artificial Dyes in the Commercial Jellys by Use of Paper Chromatography and Thin-Layer Chromatography. (Paper chromatography와 Thin-layer chromatography에 의한 시판 Jelly류의 착색료에 관한 고찰)

  • 구성회;오석흔;우세홍;한양일;이성호;남궁석;박선이
    • Journal of Environmental Health Sciences
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    • v.4 no.1
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    • pp.10-12
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    • 1977
  • A study was carried out to detect of illegal artificial dyes and to confirm the used rate of illegal dyes in the production process of commercial jellys by thindayer chromatography and paper chromatography, from March, 1976 to July, 1976. The following conclusions were obtained 1. Seperated dyes were amaranth, erythrosin, tatrazin, sunset yellow FCF, fight green SF yellowish, fast green FCF and the most frequent use of amaranth. 2. Used rate of legal dyes were 94.12% (96 samples) and illegal dyes were 5.88% (6 samples) with samples 102. 3. The average Rf value of T.L.C. were amaranth (0.92), erythrosin (0.48), tatrazin (0.83), sunset yellow FCF(0.86), fast green FCF (0.63), light green SF yellowish (0.23) and paper chromatography were amaranth (0.76), erythrosin (0.44), tatrazin(0.75), sunset yellow FCF (0.76), fast green FCF (0.86), light green SF yellowish(0.52).

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Preparative Chromatographic Separaction: Simulated Moving Bed and Modified Chromatography Methods

  • Yi Xie;Koo, Yoon-Mo;Nien-Hwa Linda Wang
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.363-375
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    • 2001
  • Chromatography has been method of choice for the separation complex biologi-cal mixtures fro analytical purpose, particularly for the last fifty years. Its use has recently been extended to preparative separation where the productivity relative to the amount of resin and sol-vent used is a matter of concern. To overcome the inherent thermodynamic inefficiency of batch chromatography, as exemplified by the partial temporal usage of the resin and dilution of the product with the solvent, chromatography has been continually modified by separation engineers. Column switching and recycling represnet some of the process modifications that have brought high productivity to chromatography. Recently, the simulated moving bed (SMB) method, which claims a high separation efficiency based on counter-current moving bed chromatography. has be-come the mainstay of preparative separation, especially in chiral separation. Accordingly, this pa-per reviews the current status of SMB along with several chromatographic modification, which may be helpful in routine laboratory and industrial chromatographic practices.

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Characterization of Vitamins in Yeast Extract using Gel Filtration, Ion Exchange Chromatography and HPLC (젤 여과, 이온 크로마토그래피와 HPLC에 의한 효모 엑기스내의 비타민의 분석연구)

  • 최인호;홍억기;강환구;김인호
    • KSBB Journal
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    • v.15 no.1
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    • pp.76-79
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    • 2000
  • Complex, ill-defined mixtures of natural origin are often used as nutrients in the production of biological products through microbial fermentation. Product yields are affected by variation in these natural products. Yeast extract is a typical example of these natural products. Since it is a mixture of amino acids, peptides and nucleic acids, its composition is not well characterized. In this study, we investigated the properties of thiamine hydrochloride, riboflavin and pyridoxine hydrochlride in yeast extract by using a gel filtration chromatography, ion exchange chromatography and high performance liquid chromatography. Yeast extract solution was fractionated by gel filtration chromatography and ion exchange chromatography, and then, each fraction was analyzed by using a high performance liquid chromatography.

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Investigation of Brassinosteroid Substances in Undmia pinnatifida (미역의 Brassinosteroid 활성물질검색)

  • 문제학;현규환박근형
    • KSBB Journal
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    • v.7 no.1
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    • pp.21-26
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    • 1992
  • In order to examine the presence of brassinosteroid substance in sea mustard(Undaria pinnatifida), leaves of sea mustard were extracted with MeOH. The extract was purified by slovent fractionation, counter current distribution, silica gel adsorption chromatography, charcoal adsorption chromatography, Bondesil chromatography, and reverse phase HPLC, successively. The activity was monitored by the rice inclination test and its prescence could be confirmed in each purification step. Although sea mustard contained a less amount of the active substance than the vegetative tissue of higher plants, brassinosteroid was clearly present endogenously in sea mustard. We acknowledge that our work is probably the first publication reporting the presence of brassinosteroid in marine algae plants.

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Isolation and Partial Purification of the Steroid 9${\alpha}$-Hydroxylase from Mycobacterium fortuitum (Mycobacterium fortuitum의 스테로이드 9${\alpha}$-하이드록실라제의 분리 및 부분정제)

  • Kang, Hee-Kyoung
    • YAKHAK HOEJI
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    • v.41 no.5
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    • pp.638-646
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    • 1997
  • The steroid 9${alpha}$-hydroxylase activity has been detected in cytosol fraction, $100,00{\times}g$ supernatant of cell free extract of Mycobacterium fortuitum. The activity was not linear with protein concentration in the assay suggesting 9${alpha}$-hydroxylase is a multicomponent enzyme. The 9${alpha}$-hydroxylase system was partially purified through fractional saturation of ammonium sulfate, strong anion exchange (Mono Q) column chromatography, gel filtration (Superose 12) column chromatography, and testosterone affinity gel chromatography. Ammonium sulfate 50~60% saturated fraction of the cytosol gave 9${alpha}$-hydroxylase activity. For further purification, the half-saturated ammonium sulfate fraction was applied to Mono Q, Superose 12, or affinity gel column. The purification factors of 9${alpha}$-hydroxylase containing fraction after Mono Q, Superose 12, and affinity gel chromatography was 13, 11, and 17 respectively.

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