• Journal of Ginseng Research
• /
• v.47 no.3
• /
• pp.366-375
• /
• 2023

Background: Ginseng contains three active components: ginsenosides, gintonin, and polysaccharides. After the separation of 1 of the 3 ingredient fractions, other fractions are usually discarded as waste. In this study, we developed a simple and effective method, called the ginpolin protocol, to separate gintonin-enriched fraction (GEF), ginseng polysaccharide fraction (GPF), and crude ginseng saponin fraction (cGSF). Methods: Dried ginseng (1 kg) was extracted using 70% ethanol (EtOH). The extract was water fractionated to obtain a water-insoluble precipitate (GEF). The upper layer after GEF separation was precipitated with 80% EtOH for GPF preparation, and the remaining upper layer was vacuum dried to obtain cGSF. Results: The yields of GEF, GPF, and cGSF were 14.8, 54.2, and 185.3 g, respectively, from 333 g EtOH extract. We quantified the active ingredients of 3 fractions: L-arginine, galacturonic acid, ginsenosides, glucuronic acid, lysophosphatidic acid (LPA), phosphatidic acid (PA), and polyphenols. The order of the LPA, PA, and polyphenol content was GEF > cGSF > GPF. The order of L-arginine and galacturonic acid was GPF >> GEF = cGSF. Interestingly, GEF contained a high amount of ginsenoside Rb1, whereas cGSF contained more ginsenoside Rg1. GEF and cGSF, but not GPF, induced intracellular [Ca²⁺]_i transient with antiplatelet activity. The order of antioxidant activity was GPF > GEF = cGSF. Immunological activities (related to nitric oxide production, phagocytosis, and IL-6 and TNF-α release) were, in order, GPF > GEF = cGSF. The neuroprotective ability (against reactive oxygen species) order was GEF > cGSP > GPF. Conclusion: We developed a novel ginpolin protocol to isolate 3 fractions in batches and determined that each fraction has distinct biological effects.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.3
• /
• pp.503-511
• /
• 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.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.44 no.5
• /
• pp.673-680
• /
• 2015

In this study, the immune-enhancing effects of purified polysaccharides from ascidian (Halocynthia roretzi) tunic were investigated. Crude polysaccharides (AP) were isolated by enzyme extraction (neutrase, $60^{\circ}C$, 15h), ethanol precipitation, and lyophilization. In addition, crude polysaccharides were further fractionated into unabsorbed fractions (APF-I, fraction No. 11~17) and absorbed fractions (APF-II, fraction No. 22~37) by DEAE-sepharose CL-6B column chromatography in order to isolate immune regulating polysaccharide. The major constituents in APF-I and APF-II were total sugar (66.62% and 27.03%), uronic acid (47.53% and 15.87%), hexosamine (16.62% and 46.79%), and protein (2.43% and 4.94%), respectively. APF-I increased production of nitric oxide (NO) and cytokines, such as tumor necrosis factor-alpha (TNF-${\alpha}$) and interleukin (IL)-6 in a dose-dependent manner. The mRNA expression levels of inducible NO synthetase, cyclooxygenase-2, TNF-${\alpha}$, and IL-6 were markedly increased as determined by polymerase chain reaction analysis. The above data led us to conclude that macrophage activation of purified polysaccharides was higher than that of crude polysaccharides. The polysaccharides isolated from ascidian tunic investigated herein are useful as natural immune enhancing agents.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.41 no.6
• /
• pp.409-413
• /
• 2008

A simple method for the purification of porphyran from laver Porphyra yezoensis was developed to obtain information for the development of food materials with biological functionality. Crude porphyran (CP) was extracted from dried laver in boiling water for 3 h, and then fractionated using cetylpyridinium chloride into an acidic fraction (CP-F1) and a neutral fraction (CP-F2). CP-F1 was fractionated further by fractional ethanol precipitation. Fraction CP-F1-70, precipitated at an ethanol concentration of 61-70% was the major fraction containing 68.1% of the yield from the initial fraction CP-F1. The CP-F1-70 fraction displayed a single band on Sepharose CL-4B with a molecular mass of 550 kDa, indicating a homogeneous polysaccharide. The molar ratio of galactose, 3,6-anhydro-L-galactose, 6-0-methyl-D-galactose and ester sulfate of CP-F1-70 was 1:0.32:0.07:0.53. This method is very useful for rapid and large-scale preparation of purified porphyran because it is compatible with mass production.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.40 no.1
• /
• pp.14-19
• /
• 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.

• The Korean Journal of Food And Nutrition
• /
• v.28 no.5
• /
• pp.821-828
• /
• 2015

ASW0 is a polysaccharide derived from the perennial herb Aster scaber Thunberg. We isolated ASW0, a fraction of crude polysaccharide, by means of ethanol precipitation and dialysis after hot water extraction to investigate its physicochemical properties and immunostimulatory effects. ASW0 contains neutral sugar (45.7%), acidic sugar (51.6%), protein (2.3%), and 2-keto-3-deoxy-D-manno-octonate (KDO) (0.4%). The neutral sugar in ASW0 (in mole percentage) was mainly composed of arabinose (34.5 mol%), glucose (31.1 mol%), galactose (14.9 mol%), and rhamnose (8.1 mol%), which are characteristic of pectic polysaccharides. ASW0 also contained small amounts of xylose, mannose, and fucose. The anti-complementary activity of ASW-0 was similar to that of polysaccharide K (used as positive control). ASW0 exhibited no cytotoxicity in RAW 264.7 macrophages and dramatically increased nitric oxide (NO) production in a dose dependent manner ($0.3{\sim}30{\mu}g/mL$). Also, macrophages stimulated with ASW0 showed enhanced production of immunostimulatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor alpha ($TNF-{\alpha}$) in a dose dependent manner. These results suggest that the ASW0 have a potent immunostimulatory effect and can be used as a natural immune health ingredient.

• Microbiology and Biotechnology Letters
• /
• v.43 no.4
• /
• pp.391-395
• /
• 2015

We examined the anti-inflammatory effect and anti-tumor activity of Tetragonia tetragonioides crude extracts and fractions. The anti-inflammatory activity of T. tetragonioides was exuded through the inhibition of lipopolysaccharide (LPS, $1{\mu}g/ml$), induced nitric oxide (NO) and interleukin (IL)-$1{\beta}$ production. The production of IL-6 and tumor necrosis factor $(TNF)-{\alpha}$ also decreased in LPS induced RAW264.7 cells after treatment with polysaccharide (PS) fraction. Furthermore, the hexane (HX) fraction strongly inhibited the granulocytes macrophage-colony stimulating factor (GM-CSF) production. In ICR mice previously inoculated with Sarcoma 180, the life prolongation effects were 16.67% with an intraperitoneal injection of methanol (MeOH) extract and polysaccharide fraction at a dose of 100 mg/kg/day. The results are an important preliminary step toward the development of effective anti-inflammatory and anti-tumor agents using T. tetragonioides.

• Applied Biological Chemistry
• /
• v.41 no.1
• /
• pp.60-66
• /
• 1998

During the screening for anti-complementary activity from 10 kinds of edible mushrooms, an alkali extract of Agaricus bisporus showed the highest activity through the complement fixation test. The crude anti-complementary material(AB-0) from Agaricus bisporus was obtained by the alkali extraction using 1 N NaOH containing 5% urea$(65^{\circ}C)$, followed by methanol reflux, dialysis and lyophilization. The fraction AB-O showed potent anit-complementary and anti-tumor activity against sarcoma-180 injected mice. The fraction AB-0 was divided into 5 fractions(AB-20, AB-40, AB-60, AB-80, AB-A) by gradual acetone precipitation. Among them fraction AB-20 having the highest activity and yield was found to contain 39% carbohydrate and 46% protein. The anti-complementary protein-bound polysaccharide AB-20 consisted of glucose, arabinose, xylose, galactose and mannose in a molar ratio of 6.49 : 1.98 : 1.24 : 1.00 : 0.71, respectively and its main component amino acids were alanine(20.59%), isoleucine(16.85%), glutamine+glutamic acid(14.12%) and leucine (13.83%). The anti-complementary activity of AB-20 was decreased greatly by periodate oxidation, but decreased slightly by pronase digestion. This indicates that polysaccharide moiety is corelated with the anti-complementary activity and that protein is also involved in the activity.

• The Korean Journal of Food And Nutrition
• /
• v.10 no.3
• /
• pp.375-381
• /
• 1997

The anticoagulant polysaccharide was screened from the ediable mushrooms. Among them, alkali extract of Ganoderma lucidum showed the highest activity in aPTT. A crude polysaccharide fraction (GL-I) was prepared from the 1N NaOH solution extract of Ganoderma lucidum followed by methanol-reflux, precipitation with ethanol, dialysis and lyophilization. GL-I inhibited the intrinsic pathway in blood coagulation pathway and exhibited concentration dependent anticoagulation effects. The anticoagulant activity of GL-I was decreased greatly by periodate oxidation, but was not changed by pronase digestion. These suggest that carbohydrate moiety may be related to the anticoagulant activity. GL-I consisted of glucose, galactose, fucose, xylose, mannose, arabinose in a molar ratio of 19.3:3.0:2.3:1.3:1.0:0.3. GL-I was partially purified on the DEAE-Toyopearl 650C(GL-IalongrightarrowGL-If) and on the Sephadex G-100(GL-Ic-ilongrightarrowGL-Ic-II).

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.33 no.10
• /
• pp.1589-1593
• /
• 2004

The aims of this study were to investigate the antioxidative and antitumor effects of crude polysaccharide fraction from Pleurtus eryngii (CPPE). CPPE inhibited autoxidation of linoleic acid at 1,000 $\mu$g/mL concentration, and the inhibitory rates of lipid peroxidation in rat liver microsome were 5.28, 9.40, and 32.5% at 10, 100, and 1,000 $\mu$g/mL concentrations, respectively. After treatment with CPPE for 72 hours, the inhibitory rates against MCF-7, A549 and AGS cell lines showed 42.3, 33.4 and 26.7% at concentration of 1,000 $\mu$g/mL, respectively. Results of CPPE treatment at 100 and 300 mg/kg/day for 7 days in sarcoma-180 bearing-mice showed survival rates of 70 and 90%, respectively. Body weights of mice treated with CPPE were significantly decreased when compared with the control.