• Title/Summary/Keyword: ethanol precipitation

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Isolation and Characterization of Arrowroot Leaf Proteins (칡잎단백질(蛋白質)의 분리(分離) 및 그 성질(性質)에 관한 연구)

  • Lee, Ka-Sun;Yim, Kook-Yi;Choi, Woo-Young;Oh, Man-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.4
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    • pp.345-352
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    • 1985
  • To characterize the protein from arrowroot leaf, proteins were extracted and separated from arrowroot leaf, then its amino acid composition and functional properties were studied. Protein in arrowroot leaf was consisted of 18.5% albumin, 33.5% globulin, 34.0% glutelin, 6.2% protamine and 7.8% insoluble residues. The rates of precipitation of proteins which extracted with water, 1M NaCl, and 0.015N NaOH as a solvent were 84.7% (at pH 3.0), 76.4% (at pH 2.5) and 86.4% (at pH 4.0), respectively. The extracted proteins were separated up to about 90% by organic solvents such as ethanol and acetone at 80% concentration, Composition of arrowroot leaf protein concentrates were: $1{\sim}2%$ moisture, $59{\sim}67%$ protein, $4{\sim}8%$ ash and $5{\sim}6%$ (dialyzed concentrates) or $1{\sim}2%$ (acetone-treated ones) lipid. Main amino acids of the concentrates were aspartic acid, glutamic acid and glycine. Solubility profile of the concentrates according to pH was typical. The minimum solubility (below pH 5.0) of acetone extracted protein concentrates was lower than that of unextracted ones, whereas the reverse was true for pH value above this region. Bulk density, water and fat absorption of the concentrates were attributable to correlation to the treatment of acetone. And the bulk density of the concentrates was negatively correlative to both water and fat absorption. Emulsifying and foaming properties were not varied with the treatment of acetone.

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Characterization and Purification of the Bacteriocin Produced by Bacillus licheniformis Isolated from Soybean Sauce (간장에서 분리한 Bacillus licheniformis가 생산하는 박테리오신의 특성 및 정제)

  • Jung, Sung-Sub;Choi, Jung-I;Joo, Woo-Hong;Suh, Hyun-Hyo;Na, Ae-Sil;Cho, Yong-Kweon;Moon, Ja-Young;Ha, Kwon-Chul;Paik, Do-Hyeon;Kang, Dae-Ook
    • Journal of Life Science
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    • v.19 no.7
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    • pp.994-1002
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    • 2009
  • A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

Purification and Characterization of Sulfated Polysaccharide Isolated from Hot Water Extract of Pachymeniopsis elliptica (Pachymeniopsis elliptica의 열수 추출물로부터 분리한 함황 다당류의 정제 및 특성)

  • Lee, Sun-Hee;Jun, Woo-Jin;Yu, Kwang-Won;Chun, Hyug;Shin, Dong-Hoon;Hong, Bum-Shik;Cho, Hong-Yon;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.1191-1197
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    • 2000
  • In the preliminary study, we investigated the anti-complementary activities of 62 extracts from Korean edible seaweeds. Of those, Pachymeniopsis elliptica showed the highest anti-complementary activity. Therefore, it was purified as follows; i) PE-1 by ethanol precipitation, ii) PE-1-C by ultrafiltration, iii) PE-1-CIV by DEAE-Toyopearl 650C, and iv) PE-1-CIV-ii by Sepharose CL-6B. The purified compound, PE-1-CIV-ii, was the complexed homogeneous polysaccharide (molecular mass: 780 kDa) with 82.9% of anti-complementary activity. Also, it contained a significant amount of sulfate group (30.5%), which indicated it as a sulfated algal polysaccharide. Its structural monosaccharides were galactose (44.3%), 3,6-anhydrogalactose (34.0%), glucose (8.2%), fucose (5.4%), xylose (5.2%) and rhamnose (2.9%). After the treatment of periodate on a sample, a significant decrease in anti-complementary activity was found, which was a characteristic of bioactive polysaccharides. And-tumor activity of PE-1-A, B and C was tested in the sarcoma-180 solid tumor model. The PE-1-C with the largest molecular mass (more than 300 kDa) showed 81% of inhibition on the solid tumors, suggesting that the anti-complementary activity was, at least in part, related to anti-tumor activity. Based upon these results, the purified polysacchardes could be an immunopotentiator in vivo.

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Purification of Complement System-Activating Polysaccharide from Hot Water Extract of Young Stems of Cinnamomum cassia Blume (계지(桂枝) 열수추출물로부터 보체계 활성화 다당의 정제)

  • Kweon, Mee-Hyang;An, Hyun-Jung;Shin, Kwang-Soon;Na, Gyeong-Su;Sung, Ha-Chin;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
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    • v.29 no.1
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    • pp.1-8
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    • 1997
  • A complement system-activating (anti-complementary) polysaccharide was purified from the hot water extract of young stems of Cinnamomum cassia Blume. Crude polysaccharide fraction (CC-1) was prepared from the hot water extract of the young stems followed by methanol-reflux, precipitation with ethanol, dialysis, and lyophilization. The anti-complementary activity of CC-1 was decreased greatly by periodate oxidation, but was not changed by pronase digestion. These suggest that carbohydrate moiety may be related to the activation of complement system. According to its ionic strength CC-1 was fractionated first using cetavlon to give 4 fractions, CC-2, 3, 4 and 5. Among them CC-2 fraction was found to retain the highest activity and yield. CC-2 was separated to an unabsorbed neutral sugar portion (CC-2-I) and seven absorbed acidic sugar fractions $(CC-2-II{\rightarrow}CC-2-VIII)$ on DEAE-Toyopearl 650C (Cl-). CC-2-III showing higher anti-complementary activity and yield than those of other fractions, was further purified on the gel permeation of Sephadex G-100 and Sepharose CL-6B to CC-2-IIIa-3. CC-2-IIIa-3 was determined to have a homogeneity hy GPC (Sepharose CL-6B) and HPLC. Gel chromatography using standard dextrans gave a value of $2.4{\times}10^5$ for the molecular weight. The purified polysaccharide, CC-2-IIIa-3 consisted of arabinose, xylose, glucose, galactose, galacturonic acid and glucuronic acid in a molar ratio of 5.56 : 3.77 : 1.87 : 1.00 : 5.12 : 3.13 and contained no nitrogen.

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Cytotoxicities of Hydrolyzed Crude Laminaran from Eisenia bicyclis on the SNU-1, HeLa and SW Cells (대황으로부터 추출한 crude laminaran 가수분해물의 암 세포독성)

  • Do, Jeong-Ryong;Kim, Dong-Soo;Park, Jong-Hyuk;Kim, Young-Myung
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.793-798
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    • 2006
  • We investigated the effects on the cytotoxicity against several cancer cells of the hydrolysis and molecular weight fractionation of crude laminaran from E. bicyclis, a brown seaweed collected from Uleung island in Korea, was extracted with boiling water and then crude laminaran was prepared by ethanol precipitation of extract obtained after elimination of calcium alginate by calcium chloride. Crude laminaran was hydrolyzed by enzyme (Econase CE), acid (0.1 N HCl) and autoclaving ($121^{\circ}C$, 180 min), and the molecular weight fractions by ultrafiltration to generate molecular weight fractions. Total sugar and sulfate contents of hydrolyzed laminaran were 72.3 and 3.5% (enzyme hydrolysate), 68.5 and 3.0% (acid hydrolysate), 70.2 and 3.2% (autoclaved), and monosaccharides of which consisted of glucose (74.7-78.5%), mannose (9.9-11.5%), galactose (8.5-9.6%) and fucose (3.1-4.5%), respectively. When the cytotoxicity of hydrolyzed laminaran on SNU-1, HeLa and SW cells was evaluated by MTT assay, growth-inhibitory activity of the enzyme hydrolysate against cancer cells was higher than that of acid hydrolysate or autoclaved laminaran. Furthermore, the fraction at a molecular weight range of 10 to 50 kDa revealed higher anti-proliferative activities. The $IC_{50}$ values of 10-50 kDa fraction at a molecular weight range of 10 to 50 kDa revealed higher anti-proliferative activities. The $IC_{50}$ values of 10-50 kDa fractions on SNU-1, HeLa and SW cells were 60.4, 58.6 and 53.9 ${\mu}g/mL$ for enzymatic hydrolysate, 75.6, 73.5 and 77.4 ${\mu}g/mL$ for acid hydrolysate, and 61.7, 68.2 and 60.8 ${\mu}g/mL$ for autoclaved, respectively.

Physiological Activities of Extract from Edible Mushrooms (주요 식용버섯 추출물의 생리활성 효과)

  • Choi, Se-Jin;Lee, Yeon-Sil;Kim, Jin-Kyung;Kim, Jin-Kyu;Lim, Soon-Sung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1087-1096
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    • 2010
  • This study was conducted to investigate the physiological activity of extracts of fresh mushrooms. The components were extracted by hot water; subsequently, the hot-water extract was subjected to 60% ethanol precipitation to yield high-molecular-weight (HMW) and low-molecular-weight (LMW) fractions. Total polyphenol contents, $\beta$-glucan contents, electron-donating ability (EDA), superoxide dismutase (SOD)-like activity, nitrite-scavenging activity, fibrinolytic activity, nitric oxide (NO) production, and inhibition of NO production of the mushroom extracts were measured using lipopolysaccharide (LPS)-stimulated murine macrophages, RAW 264.7 cells. The extracts of Lentinus edodes (Berk.) Singer and Pleurotus ostreatus (Fr.) Kummer contained the highest levels of $\beta$-glucan (33.5% and 25.57%, respectively). Further, the LMW fractions of the Phellinus linteus contained the highest levels of polyphenols (233.23 mg/g). The EDA of LMW fractions (10 mg/mL) of the Phellinus linteus and Agaricus bisporus were 80.74% and 51.35%, respectively. Further, SOD-like activities of the LMW fractions were high as compared to those of the HMW fractions. Nitrite-scavenging activities of the LMW fractions (pH 1.2; concentration, 10 mg/mL) of the Phellinus linteus and Pleurotus ostreatus (Fr.) Kummer were 75.95% and 41.05%, respectively. The fibrinolytic activity of the LMW fractions of all mushrooms showed no enzyme activity by fibrin plate assay. The fibrinolytic activity of the extracts of Tricholoma matsutake was the greatest inhibitory activity at 60.4%. Further study revealed that the mushroom extracts exhibited anti-inflammatory effects on RAW 264.7 cells. The LMW fraction ($500\;{\mu}g/mL$) of the Phellinus linteus considerably inhibited NO production (100%).

Studies on the Chemical Structure of the New Polysaccharide C - (The New Polysaccharides of Gum Tragacanth. II) - (Tragacanth gum 의 신다당류(新多糖類) C 의 화학구조(化學構造) - Tragacanth gum의 신다당류(新多糖類)에 관(關)한 연구(硏究) 제2보(第二報) -)

  • Lee, Sung-Hwan
    • Applied Biological Chemistry
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    • v.3
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    • pp.25-48
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    • 1962
  • The polysaccharide C prepared from gum tragacanth powder (U. S. P. grade) by the precipitation method with 85% ethanol was a neutral polysaccharide, $[{\alpha}]^{30}_D-72.2$. The polysaccharide C consisted of L-rhamnose, D-xylose, L-arabinose and D-galactose in the molar ratio 2:1:17:9 (Table 1, 2, 3, ). The polysaccharide C was methylated with dimethylsulphate and 40% NaOH, and Purdies regent. The hydrolyzate of fully methlated product ($[{\alpha}]^{22}_D-102$ in chloroform, the methoxy content 40.6%) was composed of 2, 3, 5-tri-O-methyl-L-arabofuranose (I), 3,4-di-O-methyl-L-rhamnopyranose (II), 2,3-di-O-methyl-D-xylose (III), 2,3,4-tri-O-methyl-D-galactopyranose (IV), 2,4-di-O-methyl-L-arabopyranose (?), 2,4-di-O-methyl-D-galactose(VI), 2-O-methyl-D-arabinose (VII), and L-arabopyranose(VIII) (Table 4, 5, and Fig. 4). The first partial hydrolysis (A) of the polysaccharide C with 0.05N-HCl for 4.5 hours at $80-85^{\circ}C$ released only L-arabinose: the second hydrolysis (B) with 0.1N-HCl for 5 hours at $80-85^{\circ}C$, L-arabinose and D-galactose; and the third hydrolysis (C) with 0.3N-HCl at $90-95^{\circ}C$ in sealed tube, L-rhamnose, D-xylose, L-arabinose and D-galactose. From the unhydrolyzate A' were found L-rhamnose, D-xylose, L-arabinose, and D-galactose; from B' L-rhamnose, d-xylose, L-arabinose and D-galactose; and from C' D-xylose and D-galactose respectively (Table 6). The periodate consumption and formic acid production of the polysaccharide C were measured at various time intervals. After 120 hours periodat was consumed by 1.23 mole per $C_5H_8O_4$ and formic acid was produced 0.78 mole per $C_5H_8O_4$ (Table 7). Although a definite chemical structure for this polysaccharide C may not be formulated, experimental data, especially, from methylation, partial hydrolysie and determination of its molar ratio, and periodate analysis showed that the polysaccharide C is a highly branched polysaccharide and would be constructed of galactoaraban as a main chain residue and L-arabofuranose, D-galactopyranosyl $(1{\rightarrow}1)$-L-arabofuranose, D-xylopyranosyl $(1{\rightarrow}2)$-L-rhamnopyranosyl $(1{\rightarrow}1)$-L-arabofuranose, and L-rhamnopyranosyl $(1{\rightarrow}1)$-arabofuranose, and D-galactopyranosyl-$(1{\rightarrow}2)$-L-arabopyranosyl-$(1{\rightarrow}1)$-I-arabofuranose as a branch chain or end group (page 21).

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Characterization and β-secretase Inhibitory Activity of Water-soluble Polysaccharides Isolated from Phellinus linteus Fruiting Body (상황버섯 자실체로부터 분리된 수용성 다당류의 특성 분석 및 이의 베타 시크리타아제 활성 저해효과)

  • Jo, Hang Soo;Choi, Doo Jin;Chung, Mi Ja;Park, Jae Kweon;Park, Yong Il
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.229-234
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    • 2012
  • A key molecule in the pathogenesis of Alzheimer's disease (AD) is the ${\beta}$-amyloid peptide ($A{\beta}$) generated by ${\beta}$-secretase activity, an aspartic protease. This study was designed to evaluate inhibitory effect of the high-molecular weight water-soluble polysaccharides (Et-P) isolated and purified from Phellinus linteus fruiting body on ${\beta}$-secretase activity. The Et-P was purified from the hot water extract of Phellinus linteus fruiting body mainly by 75% ethanol precipitation and DEAE-Cellulose column chromatography. From the DEAE-Cellulose chromato-gram and molecular weight analysis, the Et-P was shown to be a mixture of three polysaccharides with molecular mass of 1,629, 1,294, and 21 kDa, respectively. The monosaccharide composition of Et-P was determined to be glu-cose, galactose, and mannose as major sugars, glucose being the most prominent one (48% in mole percentage). The elemental analysis and FT-IR analysis suggested that Et-P is typical polysaccharides having at least partially ${\beta}$-linkages and possible existing as complex with phenolic compounds. The laminarinase digestion and HPAEC-PAD analysis suggested that Et-P is a variant of beta-(1,3)-glucans. The Et-P showed DPPH radical scavenging activity and, especially, a significant inhibitory activity on ${\beta}$-secreatase activity (48% inhibitin at 100 ${\mu}g/mL$), suggesting that they may inhibit the formation of $A{\beta}$ which is the major causative of Alzheimer's disease. The results of this study suggest that the water soluble polysaccharides of Phellinus linteus fruiting body can be a potent material for the development of preventive or therapeutic agents for AD.