• Title/Summary/Keyword: Ethanol purification

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Extraction & Purification of ${\beta}$-carotene from Recombinant Escherichia coli (재조합 대장균으로부터 고순도 베타-카로틴의 추출 및 정제)

  • Jo, Ji-Song;Nguyen, Do Quynh Anh;Yun, Jun-Ki;Kim, Yu-Na;Kim, You-Geun;Kim, Sung-Bae;Seo, Yang-Gon;Lee, Byung-Hak;Kang, Moon-Kook;Kim, Chang-Joon
    • Microbiology and Biotechnology Letters
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    • v.37 no.3
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    • pp.231-237
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    • 2009
  • This paper aimed to develop a solvent extraction and purification process to recover high-purified ${\beta}$-carotene from recombinant Escherichia coli. Cells harvested from the culture broth were treated through numerous steps: dehydration, solvent extraction, crystal formation and separation. To optimize the extracting condition, experiments were carried out to investigate the effect of cell disruption, temperature, organic solvents, solvent-biomass ratio on the yield of ${\beta}$-carotene extracted from cells. The result indicated that no significant differences of extraction yield were observed from cells with or without step of cell disruption. Among different extracting solvents, the highest extraction yield of ${\beta}$-carotene, 30.3 mg-${\beta}$-carotene/g-dry cells, was obtained with isobutyl acetate at solvent-biomass ratio 25 mL/g-dry cells at $50^{\circ}C$. Notably, in case of acetone, the extraction yield was quite low when using acetone itself, but increased almost up to the highest value when combining this solvent and olive oil. The purity of ${\beta}$-carotene crystals obtained from crystallization and separation was 89%. The purity degree was further improved up to 98.5% by treating crude crystals with additional ethanol washing.

Ethanol Extract of Hermetia illucens Larvae Inhibits Adipogenesis in 3T3-L1 Adipocytes (동애등에 유충 에탄올 추출물의 지방세포 분화 억제 효과)

  • Park, Ji Yeong;Kwak, Kyu-Won;Choi, Ji-Yeon;Lee, Si-Eun;Kim, Yong-Soon;Koo, Bonwoo;Kim, Eunsun;Park, Kwanho;Kim, Sun Young
    • Journal of Life Science
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    • v.31 no.12
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    • pp.1094-1099
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    • 2021
  • Hermetia illucens (Black soldier fly) is attracting attention as an environmental purification insect because it can supply a wide range of by-products of the agricultural food industry. Also, it has a potential feed for fish, birds, and pets due to a short life cycle and excellent nutritional components. Several pharmacological effects, including antimicrobial, of H. illucens have been reported. However, no study has focused on antiobesity effects of ethanol extract of H. illucens. In this study, we aimed to assess the anti-obesity effects of ethanol extract of H. illucens larvae (HIE) through inhibition of differentiation of 3T3-L1 preadipocytes into adipocytes. The amount of lipid accumulated in adipocytes was measured by oil red-O staining, and the inhibitory effect on adipogenesis was confirmed. The expression levels of factors related to adipocyte differentiation and fat synthesis were determined using Western blot analysis. Lipid droplet formation in adipocytes was remarkably inhibited by HIE. In addition, treatment with 400 ㎍/ml of HIE significantly reduced the expression levels of peroxisome proliferator-activated receptor γ and CCAAT/enhancer-binding protein α-transcription factors involved in adipocyte differentiation. Therefore, the results of this study indicate that HIE is a potential anti-obesity agent because it inhibits adipocyte differentiation.

Purification of Antimicrobial Compounds and Antimicrobial Effects of Schima wallichii subsp. liukiuensis against Candida sp. (Schima wallichii subsp. liukiuensis의 Candida종에 대한 항균효과 및 항균물질의 분리정제)

  • Choi, Mynug-Suk;Shin, Kuem;Yang, Jae-Kyung;Ahan, Jin-Kwon;Kwon, Oh-Woong;Lee, Yi-Young
    • KSBB Journal
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    • v.16 no.3
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    • pp.269-273
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    • 2001
  • To develop natural antimicrobial substances from Theaceae, Schima wallichii subsp. liukiuensis was selected from 218 woody plants, and antimicrobial compounds against bacteria, fungi, and yeast were isolated. The antimicrobial activity of ethanol extracts proved higher than those of other organic solvents. The antimicrobial activity of S. liukiuensis extract showed no differences in sesonal variation, but, that of plant part was high in bark at autumn. An antimicrobial substance was isolated from the extract of Schima using column chromatography packed with silica gel and sephadex LH-20, and then a purified antimicrobial substance (Compound I) was obtained using HPLC analysis. The Compound I in the analysis of UV, IR, and GC-MS presumed a triterpene or steroidal saponin, ${\alpha}$-sitisterol as aglycon combined three sugars. The minimal inhibitory concentration (MIC) of the Compound I against a bacteria, fungi, and yeast were 1.25 g/L, 5.0 g/L, and 0.040 g/L, respectively. This is much lower than the MIC of hinokitiol, an natural antimicrobial compound used commercially, which suggests that Compound I could be developed as a natural preservative and pharmaceuticals.

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Chemical Features and Purification of Immunostimulating Polysaccharides from the Fruit Bodies of Agaricus blazei (신령버섯(Agaricus blazei)으로부터 면역증강활성 다당류의 분리 및 화학적 특성)

  • Cho, Soo-Muk;Park, Jeong-Sik;Kim, Kwang-Po;Cha, Dong-Yeol;Kim, Hwan-Mook;Yoo, Ick-Dong
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.170-174
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    • 1999
  • Water-soluble polysaccharides from the fruiting bodies of Agaricus blazei Murill were extracted with 0.9% sodium chloride and hot water, successively. The purified polysaccharides showed a potent immunostimulating activity. Eight major polysaccharides, which were named from AG-l to AG-8, were fractionated and purified by ethanol precipitation, ion exchange chromatography on DEAE-cellulose and gel filtration on Toyopearl HW 65F. These polysaccharides were identified to be homogeneous by analysis of HPLC. Three major active polysaccharides (AG-2, -3, and -6) showed relatively strong immunostimulating activity. AG-2 and -3 were composed of glucose, galactose and mannose in the molar ratios of 74.0:15.3:10.7 and 63.6:17.6:12.7, respectively. AG-6 was composed of glucose and ribose in the molar ratios of 81.4:12.6.

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Purification and Biochemical Characteristics of a 45 kDa Fibrinolytic Enzyme from a Halophile (호염성균 유래 45 kDa 혈전용해효소의 순수분리와 생화학적 특성)

  • Kim, Do-Hyoung;Park, Jeong-Uck;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Choi, Yung-Hyun;Joo, Woo-Hong;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.20 no.2
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    • pp.183-189
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    • 2010
  • A fibrinolytic enzyme producing Bacillus sp. J-19 was isolated from the popular Korean seasoning, pickled anchovy. The fibrinolytic enzyme was purified to homogeneity by chromatographic methods including ethanol precipitation and gel-filtration using Sephadex G-50. Compared to the crude enzyme extract, the specific activity of the enzyme increased 1021-fold with a recovery of 23%. The purified enzyme was estimated to be approximately 45 kDa by SDS-PAGE. Especially, the amidolytic activity in the presence of the synthetic substrate for serine protease (H-D-Ile-Pro-Arg-pNA, S-2288) represented approximately 17 U/mg. In addition, more than the 60% activity of the 45 kDa fibrinolytic activity was maintained in the presence of up to 30% (w/v) sodium chloride. These findings could provide a unique fibrinolytic enzyme, leading to a potential thrombolytic agent.

Study on Cadalene Compounds Purified from Zelkova serrata Wood II - Biological activities of 7-hydroxy-3-methoxycadalene and purification of cadalene homologues - (느티나무에서 단리한 카달렌 화합물에 관한 연구 II - 7-Hydroxy-3-methoxycadalene의 생물활성 측정 및 카달렌 동족체 분리 -)

  • Choi, Joon-Weon;Mun, Sung-Hee;Choi, Don-Ha
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.1
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    • pp.78-86
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    • 2009
  • In this study 2 cadalene homologues - 3,7-dimethoxycadalene and 7-hydroxycadalene-5,8-quinone (keyakinone A)-were further identified from ethanol extracts of Zelkova serrata wood, except 7-hydroxy-3-methoxycadalene. Two biological activities-scavenging activity of hydroxy radical and cell toxicity by MTT assay-were measured with 7-hydroxy-3-methoxycadalene. The scavenging activity of hydroxyl radical of the compound was excellent and increased with its concentration. At 100 ppm hydroxyl radicals were removed completely. However, MTT assay revealed that 7-hydroxy-3-methoxycadalene showed critical toxicity to the cells. When 1 ppm of the compound was treated to the cells, cell viability was reached up to 90%, while it was reduced to 22% after treatment of 9 ppm. In 4 different Ulmaceae species, such as Ulmus davidiana, Ulmus parvifolia, Ulmus macrocarpa, Ulmus macrophylla, 7-hydroxy-3-methoxycadalene was not found at all. Instead, 7-hydroxycadalene (Mw 214), in which methoxyl group is omitted from 7-hydroxy-3-methoxycadalene, was distributed in the heartwood of 4 Ulmaceae species as major cadalene compound.

Fractionation and Electrophoretic Patterns of Rice Proteins (쌀단백질의 분획 및 전기영동)

  • Kim, Su-Il;Jo, Do-Hyun
    • Applied Biological Chemistry
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    • v.26 no.1
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    • pp.65-72
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    • 1983
  • The composition of four rice protein groups is greatly affected by the extraction conditions. The extraction amounts of albumins and glubulines primarily depended on the temperature rather than the method of extraction. The total amount of glutelins, the major components of rice storage proteins, could be extracted by a successive extraction processes, extraction with 0.5% SDS-0.1M borate buffer(pH 8.3) followed by extraction with 0.5% SDS-0.6% ${\beta}-mercaptoethanol-0.1M$ borate buffer(pH 8.3). The extracted amounts of glutelin with these solvents were 54.1 and 45% respectively. The further purification of SDS soluble glutelins was achieved by Sephadex G-150 gel column chromatography. The molecular weight of the components in four protein groups has been estimated by SDS-polyacrylamide gel electrophoresis with or without ${\beta}-mercaptoethanol.$ The comparison of albumins and globulins by starch gel electrophoresis at pH 3.1 permitted us to identify seven rice varieties. However, at pH 8.95, the specific bands for Japonica type rice varieties were observed.

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Extraction and Purification of Agar from Gelidium amansii (우뭇가사리로부터 한천의 추출 및 정제)

  • Do Jeong-Ryong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.3
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    • pp.423-427
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    • 1997
  • The effect of different treatments on the quality and yield of purified agar produced from Gelidium amansii has been studied, and the extraction condition ol agar produced from G. amansii has been examined. The contents of ash, sulfate in agar produced from g. amansii collected from different places were $2.63\~2.92\%\;and\;1.38\~1.78\%$, respectively. Yields and gel strength of agar produced from G. amansii collected from different places were $31.6\~46.8\%\;and\;496\~887g/cm^2$, respectively. It was effective to extract agar at $120^{\circ}C\;for\;2\~3hrs$. Agar was purified by D.W. washing, EDTA washing, chitosan treatment, CPC treatment, PEG treatment, ethanol precipitation, acetone precipitation and propanol precipitation. The mineral contents of agar produced from Gelidium amansii were Na (2934ppm), Ca (2472ppm), Mg (2259ppm), K (2527ppm), P1(81.1ppm), Fe (66.4ppm), Al (71.7ppm), Zn (29.7ppm) and Pb (ND: not detected), respectively.

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Isolation and Purification of Protein-bound Polysaccharides from Mycelia of Flammulina velutipes Grown on Sawdust Medium (팽나무버섯 수확후의 톱밥배지로부터 단백다당류의 분리 및 정제)

  • Ha, Hyo-Cheol;Park, Shin;Park, Kyung-Sook;Lee, Shun-Woo;Jung, In-Chang;Kim, Seon-Hee;Kwon, Yong-Il;Lee, Jae-Sung
    • KSBB Journal
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    • v.10 no.5
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    • pp.589-597
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    • 1995
  • Protein-bound polysaccahrides(PBP) were isolated, purified, and characterized from the sawdust media after harvesting the fruit body of Flammulina velutipes. The yield of the crude PBP(Fr.CA) extracted from the sawdust media, was 0.367% relative to the original sawdust media. The total sugar and protein contents of Fr.CA were 19.8% and 23.8% respectively. Using the membrane filtration, the fraction of which the molecular weight is over 300 kDa(Fr.A) was isolated from the Fr.CA and the yield was 44.6% relative to the Fr.CA. This result indicates that high molecular PBP is the dominant components of the Fr.CA. The Fr.A was separated into three fractions (Fr.A-1, Fr.A-2 and Fr.A-3) whose yields are 5.8%, 8.5% and 13.2% respectively. These fractions were further purified using gel filtration, obtaining a single peak in each fraction that considered as pure PBP Among them, the yield of Fr.A-1-${\alpha}$ was 20.9% relative to the Fr.A-1, and the molecular weight was 800 kDa. Monosaccharide components such as glucose, galactose, mannose, xylose and fucose could be detected all fractions by a HPLC analysis. Especially in the Fr.A-1-${\alpha}$ fraction, the content of glucose and galactose appeared to be high.

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Isolation and Purification of an Antitumor Metabolite from Alternaria brassicicola SW-3, the Cause of Brassica Black Leaf Spot Disease. (Phytopathogenic fungus Alternaria brassicicola SW-3가 생산하는 항암활성 물질의 분리 정제)

  • 나여정;이방숙;남궁성건;정동선
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.51-56
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    • 2002
  • An antitumor substance was purified from the culture filtrate of phytopathogenic fungus Alternaria brassicicola SW-3 isolated from soil of a chinese cabbage patch, and its characteristics were investigated. Antitumor activity of A. brassicicola SW-3 was measured by MTT assay. The cytotoxic activity against human cancer cell line was detected in the culture filtrate of A. brassicicola SW-3, but no activity found in mycelium. Antitumor substance was isolated from the culture broth by ethyl acetate extraction and purified by silica gel column chromatography. Structure of the purified compound was analyzed by the instrumental analysis such as $^1$H-NMR, $^{13}$ C-NMR and IR spectroscopy. The purified fungal metabolite of an A. brassicicola SW-3, consists of 11 carbon chain with two hydroxyl groups and two epoxides which is identical to depudecin. The $IC_{50}$/ values of the active compound identified as depudecin were $69\mu$g/mL and $57\mu$g/mL against mouse melanoma B16BL6 cell line, and human hepatoma SK-HEP1 cell line, respectively.