• 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.

• 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.

• 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.

• 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.

• 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.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.343-347
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• 2005

The polysaccharides were extracted from fruiting body, mycelia, and cell-free broth of Agaricus blazei Murill. The crude polysaccharides were obtained by the ethanol addtion. They were further purified using ion-exchange chromatography and gel chromatography. Ion-exchange chromatography using DEAE-cellulose column separated neutral and acidic polysaccharides. Neutral polysaccharides were then purified with gel filtration chromatography. For single peak obtained from gel filtration chromatography was molecular weight was measured with Sepharose CL-6B. The same procedure with acidic polysaccharides were performed to get the purified polysaccharides.

• Journal of Animal Science and Technology
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• v.47 no.6
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• pp.1025-1032
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• 2005

This study was carried out to separate ovotransferrin in chicken egg white by gel chromatography and heparin affinity chromatography. In gel filtration which was performed with 50mM Phosphate buffer (pH 7.2, 0.15M salt) at a flow rate of 2.0 ml/min, ovotransferrin and ovalbumin were eluted together in fraction number 11-16. In order to separate pure ovotransferrin, fraction No. 12-14 of them which have high concentration of ovotransferrin were concentrated and rechromatographed. However, the ovotransferrin did not separated clearly. In heparin affinity chromatography, the separation was performed with 50mM ethylaminetetraacetic acid (EDTA, pH7.2) and 50mM Phosphate buffer (pH 7.2, 0.15M salt contained) on ferrous and ferric ion saturated column at as same flow rate as gel filtration system's. Ovotransferrin and albumin were eluted together at 10-15min (fraction No.3) and 15-20min (fraction No.4), respectively. However, purified ovotransferrin was eluted at 156-165min and 165-175min (tube No.32-33) with 50 mM phosphate buffer (pH 7.2, 0.15M salt free), respectively. Heparin affinity chromatography with ferric ion saturated column was resulted in the best separation of ovotransferrin rather than separation by gel chromatography and ferrous ion saturated heparin affinity chromatography.

• 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.

• Journal of Ginseng Research
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• v.13 no.2
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• pp.254-259
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• 1989

A radioprotective ginseng protein fraction was obtained from Korean white ginseng powder by the following isolation and purification procedures: Tris-HCI buffer extraction, 70% ammonium sulfate fractionation, CM-rellulosr column chromatography, heat inactivation and Sephadex G-75 column chromatography. This fraction was further purified by Sepharose 4B and Sephadex G-150 column chromatographies. Three fractions obtained were subjected to Native-PAGE and SDS-PAGE using gradient gels and the silver staining method. Molecular weights of the native proteins and their subunits were estimated.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.45-45
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• 1993

괄루근으로부터 분리된 다당류에 대하여 continuous gel electrophoresis, SDS-Polyacrylamide gel eletrophoresis, ion exchange column chromatography, Hydroxyapatite column chromatography 및 Gel filtration등의 방법을 이용하여 다음과 같은 결과를 얻었다. 1) 황산암모늄 분별침전법에 의한 렉틴의 정제도는 초추출물의 4.85배이며 DEAE Sephadex A-50 column chromatography법에 의한 정제도는 24.17배로 나타났고, 마지막 정제단계인 Sephacryl S-200 gel filtration에 의한 정제도는 47.34배로 나타났다. 2) 정제된 렉틴의 분자량은 60,000da1ton으로 나타났다. 3) 사람의 혈액형에 따른 응집효과는 90-100％로 특이성은 없었다.