• Title/Summary/Keyword: crude and purified polysaccharide

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Inhibitory Effect of Crude Acidic Polysaccharide of Korean Ginseng on Lipolytic Action of Toxohormone-L from Cancerous Ascites Fluid (인삼의 조산성다당체 성분이 암독소호르몬-L의 지방분해 억제작용)

  • 이성동;이광승
    • Journal of Ginseng Research
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    • v.14 no.1
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    • pp.10-13
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    • 1990
  • Effect of an acidic polysaccharide fraction In Korean white and red ginseng on lipolytic action of Toxohormone-L was studied. Crude acidic polysaccharide fraction was extracted from main and lateral root of Korean white and red ginseng separately and purified several times. Inhibitory effect of crude polysaccharide fraction was determined by unit (1 unit is loft inhibition rate per Is sample). Yields of purified crude polysaccharide fraction of main and lateral root of red ginseng were 2.9 and 2.2 times higher than those of white ginseng, respectively. Inhibitory effects of main root of white and red ginseng, 11.hen final reaction concentrations of sample were 50, 100, 200, 500 $\mu$g/ml, were 37.2가 and 23.7% higher than those of lateral root of white and red ginseng. Inhibitory effect of main root of red ginseng was 2.3 times higher than that of white ginseng.

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Polysaccharide Extraction and Comparison of Free Radical Scavenging Activities from Tremella fuciformis and Auricularia auricula Fruit Body (흰목이와 흑목이 버섯의 다당추출 및 유리라디칼 소거활성 비교)

  • Kim, Hyeon-Min;Hur, Won;Lee, Shin-Young
    • Food Engineering Progress
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    • v.15 no.1
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    • pp.6-14
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    • 2011
  • The polysaccharides from fruit body of Auricularia auricula and Tremella fuciformis were extracted using hot water, and partially purified through ethanol precipitation and dialysis. Free radical scavenging activities of the crude and purified polysaccharides were examined and compared each other. Free radical scavenging activities of the partially purified polysaccharides were higher than those of crude polysaccharides. DPPH free radical, ABTS radical and SOD-like activities of partially purified polysaccharide at 1 mg/mL of concentration from A. auricula were 61.7, 9.6 and 38.9%, respectively, while those of T. fuciformis were 9.6, 5.7 and 15.3%, respectively. Results of site and non-site specific hydroxyl radical scavenging activities indicated that the partially purified polysaccharide fractions from A. auricula and T. fuciformis exhibited the hydroxyl radical scavenging effect by hydrogen donating ability and iron ion chelating ability. Also, reducing powers of A. auricula and T. fuciformis were 77.1 and 14.7% of BHT (0.1%) as standard, respectively. It was suggested that antioxidant activities of A. auricula were about 1.4~6.4 times higher than those of T. fuciformis due to different levels of polyphenol content.

Characterization and Purification of Anti-Complement Polysaccharide from Spirodela polyrhiza

  • Min, Jin-Gi;Lee, Doo-Seog;Park, Jeong-Heum;Heo, Moon-Soo;Kim, Tae-Jin;Shim, Kil-Bo;Cho, Young-Je
    • Preventive Nutrition and Food Science
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    • v.9 no.1
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    • pp.21-28
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    • 2004
  • We purified and characterized a crude polysaccharide from Spirodela polyrhiza with anti-complement activities. The crude polysaccharide fraction (SP-0) which had potential anti-complement activity was extracted in hot water for 4 hrs at 10$0^{\circ}C$. The ethanol-precipitate, the crude polysaccharide traction (SP-1), showed a potent anti-complement activity. Further purification of the crude polysaccharide (SP-1) was carried out by cetavlon, ion exchange chromatography and gel column chromatography. Among cetavlon fractions, SP-4 showed the most potent anti-complement activity. When 100 $\mu\textrm{g}$/mL of SP-4 was incubated with an equal volume of normal human serum (NHS), the TCH$_{50}$ was reduced by about 78%. When the SP-4 fraction was further purified by DEAE-Sepharose (Cl$^{[-10]}$ ), the SP-4IIa, SP-4IIb and SP-4IIc, absorbed fractions, were almost the same as the anti-complement activities of SP-4. SP-4IIc, having the greatest potential activation and the highest yield by ion exchange chromatography, was further purified by gel column chromatography on a Sepharose CL-6B column. Four polysaccharide fractions of SP-4IIc-1, SP-4IIc-2, SP-4IIc-3 and SP-4IIc-4 were obtained, consisting mainly of arabinose, rhamnose, galactose and glucose, with approximate molecular weights of about 305,000, 132,000, 64,000 and 12,000, respectively. Among these subfractions, SP-4IIc-1 had the most potent anti-complement activity. When the SP-4IIc-1 aggregate was applied to a gel column chromatography in 10 mM and 50 mM NaCl solution, the position of the peak fractions shifted to a low molecular weight region, and the molecular weight of SP-4IIc-1 decreased with increased NaCl concentration in the gel column chromatography. It was found that the self-aggregation formed spontaneously in void volume by gel column chromatography using Sepharose CL-6B in water and the self-aggregation significantly affected the anti-complement function.

Isolation, Purification and Characterization of Polysaccharides that induce in vitro Immuno-Stimulation of Macrophases derived from Liquid Culture of Cordyceps militaris

  • Kwon, Jeong-Seok;An, Hyo-Sil;Hong, Eock-Kee
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.287-292
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    • 2005
  • The crude polysaccharides(C-CPF, C-CPM, C-CPB) derived from fruiting body, mycelia and mycelia free broth of cordyceps militaris were obtained by ethanol precipitation of hot water extracts. After a batch fermentation of C. militaris was carried out in a 5 L jar vessel, endo-polysaccharide and exo-polysaccharide were obtained. They were demonstrated as the hetero polysaccharides which were composed of glucsose, galactose and mannose by performed with HPAEC(high pH anion exchange chromatography) and conformation of random coil by its complex forming ability with congo red reagent. They were purified by ion exchange (DEAE-cellulose) and gel filtration chromatography. They were monitered by phenol-sulfuric acid method and Bradford method. The NO induction activities of crude polysaccharides and purified polysaccharides derived from mycelia free broth were enhanced rather than LPS(lipo polysaccharide) which was used as a general NO inducer. These effects presumably contibute to the antitumor activities. The homogenieties and molecular weights of polysaccharides were determined by using Sepharose CL-6B. The yield, molecular weights and NO induction activities of C-CPFN Fr.III, C-CPMN Fr.III, C-CPBN Fr.II were 0.387, 0.408 and 0.153, 127 K 210 K and 36 K, 40.79%, 88.72%, and 104.17%, respectively.

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Studies on the Blood Anticoagulant Polysaccharide Isolated from Hot Water Extracts of Hizikia fusiforme (톳 열수추출물로부터 분리한 혈액 항응고성 다당류에 관한 연구)

  • 양한철;김경임;서혜덕;이현순;조홍연
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.6
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    • pp.1204-1210
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    • 1998
  • This study was focused on the purification, characterization and promotion mode of an anticoagulant polysaccharide from Hizikia fusiforme. The anticoagulant crude polysaccharide(HF 0) was obtained by using hot water extraction at 100oC for 3 hrs after homogenizing desalted Hizikia fusiforme. The anticoagulant polysaccharide(HF 2 3 1a) was purified from the crude extract(HF 0) through stepwise gradient ethanol precipitation(HF 2), DEAE Toyopearl 650C(HF 2 3), Sephadex G 75(HF 2 3 1), Sepharose CL 6B(HF 2 3 1a) chromatography and HPLC to homogeneity. HF 2 3 1a was estimated at 5.3$\times$105 Da molecular weight and composed of fucose(51.92%), galactose(19.34%), mannose(13.92%), xylose (7.14%), arabinose(3.95%) and rhamnose(3.78%), and comprimised 29.7 % sulfate residue. The sulfated anticoagulant polysaccharide from HF 2 3 1a was proposed to inhibit via the intrinsic pathway and common pathway in the blood coagulation. The HF 2 3 1a exhibited the anticoagulant activity by activating an antithrombin III and the activity depended on the concentration of HF 2 3 1a. Acute toxicity of HF 2 in mice was not detected. Only 14 of 33 control mice(11.4%) that had taken saline survived for 30 min after injecting thrombin(100 NIH unit/ml).

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Isolation and Purification of Polysaccharide from Fruiting body and Culture Broth of Agaricus blazei Murill

  • Youm, Yong-Soo;Hong, Eock-Kee
    • 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.

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Screen of Functional Activity of Polysaccharide and Glycosaminoglycan from Sea Hare (Aplysia kurodai) by Cell Line (세포주를 이용한 군소 다당류와 Glycosaminoglycan의 기능성 검색)

  • Hong, Yu-Mi;Park, Si-Hyang;Yoon, Bo-Yeong;Choi, Byeong-Dai;Choi, Yeung-Joon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.14-19
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    • 2011
  • In this study, we extracted a whole polysaccharide fraction from the sea hare, Aplysia kurodai, and screened its functional properties using cell lines. The functionalities of polysaccharide and glycosaminoglycan (GAG) were investigated with RAW 264.7 cell lines. The crude polysaccharides and GAG purified DEAE-Sepharose chromatography did not show the toxicity on RAW 264.7 cell line in the range of $10\sim200{\mu}g$/mL, whereas they increased the cell growth rate. The crude polysaccharides and purified GAG also increased the production of NO, interleukin-6 and tumor necrosis factor-$\alpha$ on RAW 264.7 cell. Particularly, the purified GAG inhibited the proliferation of stomach cancer cell line, AGS, up to 40% for 72 hr incubation, but not the intestinal epithelial IEC-6 cell lines.

Structural and Physiological Characteristics of Rhamnogalacturonan II from Fruit Wines

  • Park, So-Yeon;Shin, Kwang-Soon
    • Food Science and Biotechnology
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    • v.16 no.2
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    • pp.270-274
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    • 2007
  • To characterize the polysaccharides which exist as soluble forms in fruit wines, crude polysaccharides were isolated from red, white, raspberry, wild grape, and pear wine, respectively. Among them, the crude polysaccharide (RW-0) in red wine showed the highest yield and considerable amounts of thiobarbituric acid (TBA)-positive materials. The pectic polysaccharide RW-2 was purified to homogeneity from RW-0 by subsequent size-exclusion chromatography using Sephadex G-75 and its structure was characterized. RW-2 consisted of 14 different monosaccharides which included rarely observed sugars in general polysaccharides, such as 2-O-methyl-fucose, 2-O-methyl-xylose, apiose (Api), 3-C-carboxy-5-deoxy-L-xylose (aceric acid, AceA), 3-deoxy-D-manno-2-octulosonic acid (Kdo), and 3-deoxy-D-lyxo-2-heptulosaric acid (Dha). Methylation analysis indicated that RW-2 comprised at least 20 different glycosyl linkages such as 3,4-linked fucose, 2,3,4-linked rhamnose, 3'-linked apiose, and 2,3,3'-linked apiose, being characteristic in rhamnogalacturonan II (RG-II). High performance size-exclusion chromatography indicated that RW-2 mainly comprised RG-II of higher molecular weight (12,000), and that the changes of molecular weight to apparent 7,000 under less than pH 2.0 were observed. These analyses indicated that the higher molecular weight polysaccharide in RW-2 was mainly present as a RG-II dimer.

Evaluation of Biomolecular Interactions of Sulfated Polysaccharide Isolated from Grateloupia filicina on Blood Coagulation Factors

  • Athukorala, Yasantha;Jung, Won-Kyo;Park, Pyo-Jam;Lee, Young-Jae;Kim, Se-Kwon;Vasanthan, Thava;No, Hong-Kyoon;Jeon, You-Jin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.503-511
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    • 2008
  • An edible marine red alga, Grateloupia filicina, collected from Jeju Island of Korea was hydrolyzed by cheap food-grade carbohydrases (Viscozyme, Celuclast, AMC, Termamyl, and Ultraflo) to investigate its anticoagulant activity. Among the tested enzymatic extracts of G. filicina, a Termamyl extract showed the highest anticoagulant activity. Anion-exchange chromatography on DEAE-cellulose and gel-permeation chromatography on Sepharose-4B were used to purify the active polysaccharide from the crude polysaccharide fraction of G. filicina. The purified sulfated polysaccharide (0.42 sulfate/total sugar) showed ${\sim}1,357kDa$ molecular mass and was comprised mainly of galactose(98%) and 1-2% of glucose. The sample showed potential anticoagulant activity on activated partial thromboplastin time (APTT) thrombin time (TT) assays. The purified G. filicina anticoagulant (GFA) inhibited the coagulation factor X (92%), factor II (82%), and factor VII (68%) of the coagulation cascade, and the molecular interaction (protein-polysaccharide) was highly enhanced in the presence of ATIII (antithrombin III). The dissociation constant of polysaccharide towards serine proteins decreased in the order of FXa (58.9 nM) >FIIa (74.6 nM) >FVII (109.3 nM). The low/less cytotoxicity of the polysaccharide benefits its use in the pharmaceutical industry; however, further studies that would help us to elucidate the mechanism of its activity are needed.

The Effect of Nutrients on the Production of Anti-Complementary Polysaccharide by Flammulina velutipes (Flammulina velutipes 항보체 활성다당의 생산에 미치는 영양원의 효과)

  • Lee, Hyun-Kyung;Shin, Kwang-Soon;Song, Chi-Hyeun;Sung, Ha-Chin;Yang, Han-Chul
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.360-367
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    • 1994
  • The effect of various nutrients for the production of the anti-complementary polysaccha- ride by Flammulina velutipes, was examined and the partial purification of the polysaccharide was carried out. The culture conditions and medium compositions considerably influenced the anti-complementary activity of the polysaccharides. When a culture was carried out at 23$\circ$C and pH 5.5 for 6 days in the synthetic medium supplemented with galactose and NaNO$_{3}$, the production of the anti-complementary polysaccharide was maximized. The crude polysaccharide, FV-1 was isolated from the culture broth and partially purified into two fractions, FV-1-II and FV-1-III by gel filtration using Sephadex G-100. FV-1-II was mainly consisted of xylose, mannose, galactose and glucose, and rhamnose, mannose and galactose, for FV-1-III. Also, the anti-complementary activity of FV-1-III was reduced partially in the absence of the Ca ion. When crossed immunoelect- rophoresis using anti-human C3 serum was carried out after incubation of normal human serum with the FV-1-III in the Ca ion free condition, a cleavage of C3 precipitin line was observed. These results indicate that the mode of complement activation by polysaccharide from Flammulina velutipes is via not only the classical pathway but also the alternative pathway.

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