Sepharose CL-6B Chromatography Research Articles

Screening of sweet potatoes varieties, physicochemical and biological activity studies of glycoproteins of selected varieties

In order to meet the nutritional and health needs of different consumers, the main nutrients of eight new sweet potatoes (SPs) varieties were to characterize by principal component analysis. The watermelon red (WR) variety is proving to be the preferred SPs variety because of its high nutritional value and unique sensory features. Three homogeneous fractions SPFG-1, SPFG -2 and SPFG-3 with yield of 53.02% ± 3.20%, 10.50% ± 0.61% and 5.20% ± 0.35% were obtained from the WR by multifrequency ultrasonic extraction (MUE), isolated and purified with DEAE Sepharose CL-6B and Sephadex G-100 gel chromatography. Physiochemical and structural characteristics of three fractions were studied with chemical components analysis, HPLC, HPGPC, and β-elimination, UV, FT-IR and CD spectrum analysis. SPFG-1, SPFG-2 and SPFG-3 were all conjugates with O-glycosylation with the molecular weight of 30.42 ± 2.12, 391.15 ± 25.12 and 6.85 ± 0.78 kDa, including seven monosaccharides and 17 amino acids with quite different mole percentages, that they were predominantly α-helix, β-folding and irregularly curled with a relatively small β-turning content. Meanwhile, the three conjugates exhibited excellent antioxidant and hypoglycemic activities by radical scavenging, α-glycosidase and α-amylase inhibition assays. These findings are helpful to promote the breeding, cultivation and promotion of new SPs varieties with higher nutritional value, as well as the development and application of glycoproteins.

Chondroitin sulfate is not digested at all in the mouse small intestine but may suppress interleukin 6 expression induced by tumor necrosis factor-α

Salmon nasal cartilage proteoglycan (PG) was orally administered to mice. The PG digest was recovered from the small intestine, and its sugar chain size and unsaturated disaccharide content were examined. The elution position of the PG digest following Sepharose CL-4B chromatography was consistent with that of actinase-digested PG prior to administration. The PG digest was incubated with chondroitinase ABC, which resulted in the elution pattern of the unsaturated disaccharides being identical to that of the degraded product of actinase-digested PG. The core protein of PG was digested in the mouse small intestine, but chondroitin sulfate, which is the sugar chain of PG, was not degraded at all. Then, the effects of chondroitin 4- and 6-sulfates on human colon cancer cells were examined. These chondroitin sulfates were found to suppress the expression of interleukin-6 induced by TNF-α. Overall, the chondroitin sulfate chain may act on the intestinal epithelium and suppress inflammation of the intestinal tract.

Effects of an Armillaria mellea Polysaccharide on Learning and Memory of D-Galactose-Induced Aging Mice.

Arm illaria mellea has been known and used in traditional medicine in East Asia for hundreds of years. It has already been reported that A. mellea extracts have various pharmacological effects, and the polysaccharides of A. mellea exhibit antioxidant and anti-apoptotic activities. In this study, a water-soluble polysaccharide (AMP-N-a-1), with an average molecular weight of 17 kD, was isolated and purified from the water extract of A. mellea using DEAE-52, Sepharose CL-4B, and Sephadex G-100 column chromatography. AMP-N-a-1 was mainly composed of Man (1.65%), Glca (1.64%), Rha (1.82%), Gala (2.49%), Glc (90.48%), Gal (0.89%), Xyl (0.42%), and Ara (0.61%). AMP-N-a-1 was used to study the effect on the learning and memory of mice and its underlying mechanisms. The results showed that AMP-N-a-1 could significantly increase the activities of catalase (CAT) and superoxide dismutase (SOD) and reduce the content of nitric oxide (NO) in mouse brain tissue. Meanwhile, AMP-N-a-1 could reduce the contents of norepinephrine (NE) and dopamine (DA) but could increase the content of 5-hydroxytryptamine (5-HT) in mouse brain tissue. In addition, the immunofluorescence experiment showed that AMP-N-a-1 could promote the proliferation of hippocampal dentate gyrus neurons. The above results indicate that AMP-N-a-1 can significantly improve the learning and memory of mice, and the mechanism may be that AMP-N-a-1 can participate in the regulation of learning and memory through a variety of ways.

Immunomodulatory effects of four polysaccharides purified from Erythronium sibiricum bulb on macrophages.

Four neutral polysaccharides (ESBP1-1, ESBP1-2, ESBP2-1 and ESBP3-1) were successfully purified from the water extracted crude polysaccharides of Erythronium sibiricum bulbs through the combination of DEAE Sepharose CL-6B and Sephadex G-100 chromatography; their average molecular weights were 1.3 × 104, 1.7 × 104, 9.4 × 105 and 4.1 × 105 Da, respectively. Monosaccharide component analysis indicated that ESBP1-1 and ESBP1-2 were mainly composed of glucose (Glc). ESBP2-1 was composed of Glc, galactose (Gal) and arabinose, with a molar ratio of 24.3:1.1:1, whereas ESBP3-1 comprised Glc and Gal at a molar ratio of 14.8:1. In-vitro study showed that all of the four polysaccharides were able to considerably promote the proliferation and neutral red phagocytosis of RAW 264.7 macrophage cell. They could also stimulate the production of the cell lines' secretory molecules [nitric oxide, tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β)] in a dose-dependent manner. However, ESBP1-2 was not included in IL-1β. Overall, these results suggested that polysaccharides from E. sibiricum bulbs can be developed as immunomodulatory ingredients for complementary medicines or functional foods. However, further animal or clinical studies are required.

Purification and characterization of an esterase from larval Diplolepis fructuum (Rübsaamen, 1895) (Hymenoptera: Cynipidae)

Diplolepis fructuum (Rübsaamen, 1895) (Hymenoptera: Cynipidae) is one of the important insect species that causes damages on Rosaceae species. With this study commenced in 2018 at the laboratory of Department of Biochemistry, Faculty of Science, Cumhuriyet University to get a biochemical data, an esterase (EC 3.1.1.X) from the larvae of D. fructuum was purified using Q Sepharose anion exchange, phenyl Sepharose CL-4B and Sephacryl S100 HR gel filtration chromatography, respectively. The enzyme had 6.94 U/mg protein specific activity, about 29-fold purity, and 8.8% yield. Only one activity band was observed in native-PAGE studies. The molecular weight of the esterase was estimated as 60 kDa using native-PAGE and SDS-PAGE techniques. By the kinetic data, optimum temperature and pH for the enzyme was determined as 40ºC and 9.0, respectively. The enzyme was stable for 4 h at 40ºC and pH 8.0. K m and V max values were found to be 0.035 mM and 1.41 µmol/mL.min., using 4-nitrophenyl butyrate (p-NPB) as substrate. The enzyme exhibited its highest activities on p-NPB (100%) and 4-nitrophenyl acetate (52%). All of these data indicate that the enzyme might be a typical esterase with different kinetic properties and molecular weight than esterolytic enzymes reported from other insect species.

Characterization and Antioxidant Activity of the Exopolysaccharide Produced by Bacillus amyloliquefaciens GSBa-1.

The exopolysaccharide (EPS) produced by Bacillus amyloliquefaciens GSBa-1 was isolated and purified by ethanol precipitation, and DEAE-cellulose and Sepharose CL-6B chromatographies. The molecular mass of the purified EPS was determined to be 54 kDa. Monosaccharide analysis showed that the EPS was composed of predominantly glucose, and it was further confirmed by NMR spectroscopy to be α-glucan that consisted of a trisaccharide repeating unit with possible presence of two α-(1→3) and one α-(1→6) glucosidic linkages. Microstructural analysis showed that the EPS appeared as ellipsoid or globose with a smooth surface. The EPS had a degradation temperature at 240°C. Furthermore, the EPS had strong DPPH and hydroxyl radical scavenging activities, and moderate superoxidant anion scavenging and metal ion-chelating activities. This is the first characterization of a glucan produced by B. amyloliquefaciens with strong antioxidant activity. The results of this study suggest the potential of the EPS from B. amyloliquefaciens GSBa-1 to serve as a natural antioxidant for application in functional products.

Identification, characterization and immune response of prophenoloxidase from the blue swimmer crab Portunus pelagicus and its antibiofilm activity

Prophenoloxidase is a conserved Cu-containing enzyme acting as a major defense molecule in the immune response of crustaceans. In the present research, we purified prophenoloxidase from the haemolymph of Portunus pelagicus (Pp-proPO) by Blue Sepharose CL-6B chromatography. Pp-proPO exhibited only one band with molecular weight of 75kDa on SDS-PAGE. The purified Pp-proPO was characterized through X-ray diffraction (XRD) and high-performance liquid chromatography (HPLC). Pp-proPO showed phagocytic activity on the yeast Saccharomyces cerevisiae as well as encapsulation on sepharose CL-6B beads associated with CM sepharose and beads of sodium alginate. Pp-proPO also led to strong agglutination on human erythrocytes. Furthermore, Pp-proPO showed magnified PO activity when altered with activated particles acting as pathogen combined molecular patterns (PAMPs), metal ions or other chemicals. Pp-proPO showed relevant antibiofilm activity on Gram negative bacteria Pseudomonas aeruginosa and Escherichia coli. Overall, the above results allowed us to claim that Pp-proPO play a key role in immune defense mechanisms of P. pelagicus crabs, in particular towards microbial pathogens; notably we added basic information to the functional characterization of Pp-proPO, as well as to understand its immunological role in crustaceans defense systems.

The adolescent idiopathic scoliotic IVD displays advanced aggrecanolysis and a glycosaminoglycan composition similar to that of aged human and ovine IVDs.

The present study was designed to ascertain how altered biomechanics in adolescent idiopathic scoliotic (AIS) intervertebral discs (IVDs) affected tissue compositions and aggrecan processing compared to age matched and aged human IVDs. Newborn, 2- and 10-year-old ovine IVDs were also examined. Aggrecan populations were separated by Sepharose CL2B chromatography, composite agarose polyacrylamide gel electrophoresis (CAPAGE) and identified by immunoblotting. The KS and CS content of IVD tissue extracts from AIS IVDs were compared with age-matched normal adolescent IVDs and with old human IVDs. Extracts from newborn, 2- and 10-year-old ovine IVDs were also examined in a similar manner. Adolescent idiopathic scoliotic IVD Aggrecan populations shared similar levels of polydispersity and aggregatability with hyaluronan as old IVD proteoglycans. CAPAGE demonstrated three aggrecan populations in AIS, aged human and ovine IVDs increased polydispersity and mobility in CAPAGE. AIS IVDs had GAG compositions similar to aged human and ovine IVDs. Sulphated KS (5-D-4) and chondroitin-6-sulphate, 3-B-3(+) were markers of tissue maturation, and chondroitin-4-sulphate, 2-B-6(+) was prominent in immature IVDs but its levels were lower in mature IVDs. Sulphated KS and 3-B-3(+) CS were prominently associated with IVD maturation and AIS IVDs, while the 2-B-6(+) CS isomer was associated with immature IVD tissues. The polydispersity of aggrecan in AIS IVDs, which was similar to in old human and ovine IVDs, reflected altered processing in the AIS IVDs in response to the biomechanical microenvironments the disc cells were exposed to in AIS IVDs. These slides can be retrieved under Electronic Supplementary Material.

Isolation, purification, characterization and immunostimulatory activity of polysaccharides derived from American ginseng

In this study, crude American ginseng polysaccharide (AGPS) was extracted with hot water and preliminarily purified by using resin S-8 and Polyamide columns. Then, it was further purified and separated by DEAE-Sepharose CL-6B and Sepharose CL-6B chromatography, respectively. Five main fractions were obtained, named WPS-1, WPS-2, SPS-1, SPS-2 and SPS-3. Their homogeneities and structural characteristics were elucidated based on UV–vis spectroscopy, High Performance Gel Filtration Chromatography (HPGFC), Gas Chromatography (GC), Scanning Electron Microscopy (SEM), Infrared Spectrum (IR), and NMR Spectroscopy methods. Furthermore, the immunostimulatory effects of these fractions upon splenic lymphocyte proliferation, macrophage phagocytosis and nitric oxide (NO) production, were investigated in vitro. The results indicated that their stimulations could be ordered as SPS-3>SPS-1>CPS (crude polysaccharides)>WPS-1>WPS-2>SPS-2. Among them, SPS-3 showed more potent immunomodulatory activity and could be explored as a potential immunopotentiating agent for use in functional food or medicine.

Structure, physical property and antioxidant activity of catechin grafted Tremella fuciformis polysaccharide

In this study, structural characterization, physical property and antioxidant activity of catechin grafted Tremella fuciformis polysaccharide (catechin-g-TPS) were investigated. Crude polysaccharides were isolated from the fruit bodies of T. fuciformis and further purified on DEAE-52 and Sepharose CL-4B chromatography to afford a main purified fraction (named TPS). The molecular weight of TPS was determined as 5.82×105Da by HPLC. Then, the free radical mediated grafting of catechin onto TPS was achieved by using a redox system. As compared with the unmodified TPS, catechin-g-TPS showed new bands within the range of 1300–1600cm−1 in FT-IR spectrum, and exhibited new signals at around δ 6.00 and 6.80ppm in 1H NMR spectrum. Thermogravimetric analysis indicated the thermal stability of catechin-g-TPS was higher than TPS. X-ray diffraction spectrum of catechin-g-TPS exhibited two sharp narrow diffraction peaks at 14.2 and 32.1°, corresponding to the crystalline peaks of catechin. Scanning electron microscopy observation revealed the surface of TPS was smooth, whereas the surface of catechin-g-TPS was much rough. These results all confirmed the successful grafting of catechin onto TPS. Moreover, catechin-g-TPS had higher 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power as compared to TPS.

English

Phenoloxidase (PO) is a key factor in insect immunity. On invasion of microorganisms and pathogens, prophenoloxidase (PPO) changes to its active form, PO. The present study has been conducted to purify and characterize the PO from the haemolymph of desert locust, Schistocerca gregaria (Forskal) following activation of immune system by invasion of bacteria, Bacillus thuringiensis kurstaki (Bt). PO is purified by a combination of ammonium sulfate precipitation, blue sepharose CL-6B and phenyl sepharose CL-4B chromatography yielded a 209.97-fold purity and 54.75% recovery of activity. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) reveals that the molecular weight of the purified PO is 70.154 kDa. The purified PO is characterized in terms of its biochemical and enzymatic properties by using L-DOPA as a specific substrate. Ca2+ and Cu2+ significantly stimulated PO activity when compared with other metals. The PO reaction was strongly inhibited by phenylthiourea and thiourea, moderately inhibited by ethylene diamine tetractic acid (EDTA) and poorly inhibited by ethylene glycol tetraacetic acid (EGTA) and diethyl dithiocarbamate (DTC). Inhibition of PO showed excellent recovery ability by addition of Ca2+ on EGTA-inhibited enzyme. Therefore, PO is most probably a kind of tyrosinase-type Ca2+-containing metalloenzyme. The content of Ca2+ is higher than other trace metal elements. The reactive intermediates yielded by PO with its specific substrate L-DOPA had a broad-spectrum bactericidal activity against Gram +ve bacteria (Bacillus cereus and Staphylococcus aureus) with a greater degree more than Gram-ve bacteria (Escherichia coli and Pseudomonas aeruginosa). From the present study, PO from S. gregaria is most probably a tyrosinase-type calcium-containing mono-phenoloxidase, which functions not only as a catalytic enzyme in melanin production in locusts, but perhaps also as a humoral factor in host defense via melaninization as in other insects. Key words: Schistocerca gregaria, phenoloxidase, purification.

Therapeutic efficacy of a polysaccharide isolated from Cordyceps sinensis on hypertensive rats

This study was aimed to investigate the antihypertensive effect of a polysaccharide fraction from Cordyceps sinensis on spontaneously hypertensive rats (SHR). The CSP1, one component of Cordyceps sinensis polysaccharides (CSP), was obtained after water extraction, deproteinization, de-colorization and purification with DEAE-cellulose 52. And a more homogeneous component CSP1-2 was obtained using Sepharose CL-6B chromatography. CSP1-2 mainly consisted of mannose, glucose and galactose in a molar ratio of about 2:2:1 and its average molecular weight was approximately 2.70×104Da. Pharmacological tests showed that CSP1, in which the CSP1-2 was its main component, had antihypertensive effect by stimulating the secretion of vasodilator NO, decreasing the level of ET-1, epinephrine, noradrenaline and angiotensin II, inhibiting the increase of transforming growth factor β1 (TGF-β1) and lowering the level of inflammatory mediator of C-reactive protein (CRP). These results suggested that CSP1 may possess high potential in treating hypertension.

Purification, characterization and biological activity of a novel polysaccharide from Inonotus obliquus

A novel water-soluble polysaccharide IP3a was successfully isolated and purified from I. obliquus by DEAE-cellulose, Sepharose CL-6B and Sephadex G-200 column chromatography. Chemical characterization and antitumor and immunoregulatory activity of IP3a were investigated. IP3a consisted of rhamnose, arabinose, glucose and galactose in a molar ratio of 2.5:4.6:1.0:2.6 with an average molecular weight of 48,820Da. IP3a exhibited no significant antitumor activities in vitro. However, IP3a could not only inhibit the growth of transplantable Jurkat tumor in mice significantly, but also could enhance the splenocyte proliferation and lymphocyte proliferation induced by ConA and LPS in a dose-dependent manner. At the same time, IP3a could promote cytokine secretion (IL-2, IL-6, IL-12 and TNF-α) and macrophage phagocytosis in mice. In addition, IP3a could increase Bax expression and inhibit Bcl-2 expression significantly. These results suggested that antitumor mechanisms of IP3a might be associated with improving immune response in vivo and inducing apoptosis of tumor cells in vitro. IP3a might be utilized as a potential therapeutic agent against lymphoma cancer with immunomodulatory activity.

Extraction, characterization and in vitro antioxidant activity of polysaccharides from black soybean

Optimization of extraction conditions, preliminary characterization and in vitro antioxidant activity of polysaccharides from black soybean (BSPS) were investigated. The results of Box-Behnken design showed that the optimal extraction conditions for BSPS were as follows: ratio of water to material of 20ml/g, extraction time of 6.4h and extraction temperature of 92°C, with a corresponding yield of 2.56%. The crude BSPS were further fractionated on DEAE-52 and Sepharose CL-4B chromatography to afford three purified fractions (BSPS-1, BSPS-2 and BSPS-3). Chemical analysis showed that the three purified fractions were mainly composed of carbohydrate and uronic acid. In addition, BSPS-1 was composed of arabinose, rhamnose, galactose, glucose and mannose in the molar ratio of 1.79:1.00:2.59:26.54:1.01. BSPS-2 was composed of arabinose, rhamnose, xylose, galactose and mannose in the molar ratio of 8.10:4.80:9.15:13.38:1.00. BSPS-3 was composed of arabinose, rhamnose, galactose and mannose in the molar ratio of 16.80:3.60:33.66:1.00. The results of Fourier transform-infrared spectroscopy further confirmed the characteristic polysaccharide structures of the three purified fractions. Moreover, antioxidant assays showed crude BSPS and its purified fractions had potential superoxide anion and DPPH radical scavenging activities, and their antioxidant activity decreased in the order of crude BSPS>BSPS-3>BSPS-2>BSPS-1.

Improved production, purification and bioactivity of a polysaccharide from submerged cultured Ganoderma lucidum.

Polysaccharides from Ganoderma lucidum showed multiple biological activities, such as immuno-modulating, antitumor, antioxidant, and hepatoprotective activity, etc. Adlay oil was added into the media to enhance polysaccharide production by submerged culture of G. lucidum in this work. The results revealed the optimal concentration of adlay oil was 1.5 % for polysaccharide production of G. lucidum. Analysis of the polysaccharide components confirmed that no novel components were biosynthesized by the addition of adlay oil. The main fraction of extracellular polysaccharide, GLEP-2, was isolated from the submerged culture broth of G. lucidum by ethanol precipitation, filtration, DEAE cellulose-52 and Sepharose CL-6B chromatography. GLEP-2, which was composed of glucose, galactose, mannose, arabinose, and rhamnose in a ratio of 332:55:32:13:3 respectively, had an average molecular weight of ~2.08 × 10(4) Da. The bioactivity tests demonstrated that GLEP-2 enhanced the T lymphocyte proliferation significantly at a concentration of 200 μg/mL and B lymphocyte proliferation at lower concentrations of 50 μg/mL. The results suggest polysaccharides from the submerged culture of G. lucidum are potential candidates for further development and possible commercial applications, especially in the pharmaceutical and functional foods industries.

Purification and Characterization of Polyphenol Oxidase From Agaricus bisporus

A polyphenol oxidase was isolated from Agaricus bisporus using ammonium sulfate precipitation, combined with DEAE–Sepharose Fast Flow chromatography and Phenyl Sepharose CL-4B chromatography. The molecular weight of the purified polyphenol oxidase was determined to be about 43 kDa with N-terminal sequence Ala-Thr-Asn-Ser-Gly-Thr-Leu-Ile-Ile-Phe by Edman degradation. The optimum temperature and pH for the polyphenol oxidase activity was 20°C and a pH of 6.5–7.0. Moreover, it was stable at 30 and 40°C over a 60 min pre-incubation time period. The Km and Vmax values of this polyphenol oxidase for catechol were 0.67 mM and 3333 U/ml min−1, respectively. In addition, following the cross linking with polyphenol oxidase, the emulsifying activity index, foam capacity, and foam stability of the whey proteins were evaluated to be modified. This work was useful to better understand polyphenol oxidase from A. bisporus, and may lead to its practical use in the future.

Glyceraldehyde-3-phosphate Dehydrogenase from Chironomidae Showed Differential Activity Towards Metals

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is known to interact with different biomolecules and was implicated in many novel cellular activities including programmed cell death, nuclear RNA transport unrelated to the commonly known carbohydrate metabolism. We reported here the purification of GAPDH from Chironomidae larvae (Insecta, Diptera) that showed different biologic activity towards heavy metals. It was inhibited by copper, cobalt nickel, iron and lead but was activated by zinc. The GAPDH was purified by ammonium sulphate fractionation and Chelating Sepharose CL-6B chromatography followed by Blue Sepharose CL-6B chromatography. The 150-kDa tetrameric GAPDH showed optimal activity at pH 8.5 and 37°C. The multiple alignment of sequence of the Chironomidae GAPDH with other known species showed 78 - 88% identity to the conserved regions of the GADPH. Bioinformatic analysis unveils substantial N-terminal sequence similarity of GAPDH of Chironomidae larvae to mammalian GADPHs. However, the GADPH of Chironomidae larvae showed different biologic activities and cytotoxicity towards heavy metals. The GAPDH enzyme would undergo adaptive molecular changes through binding at the active site leading to higher tolerance to heavy metals.

Purification and Partial Characterization of an Acidic α-Glucan–Protein Complex from the Fruiting Body ofPleurotus sajor-cajuand Its Effect on Macrophage Activation

The aim of this study was to purify an acidic α-glucan-protein complex from the fruiting bodies of Pleurotus sajor-caju by using the cell wall-degrading enzymes, xylanase and cellulase. The acidic glucan-protein complex was separated from a polysaccharide extract by using DEAE Toyopearl 650M anion-exchange and Sepharose CL-6B chromatography. Its homogeneity was ensured by high-performance size-exclusion chromatography and agarose gel electrophoresis. The acidic glucan-protein complex had a molecular weight of approximately 182 kDa. Fourier transform infrared spectroscopy of the acidic glucan-protein complex revealed an α-glycosidic bond and the typical characteristics of polysaccharides and proteins. The amino acid composition of the protein moiety was dominated by proline, glycine, glutamic acid and aspartic acid, indicating that the protein was highly flexible and had a negative charge. Atomic force microscopy proved that the acidic α-glucan-protein complex existed in a spherical conformation. The acidic α-glucan-protein complex stimulated the activation of macrophages, including the production of nitric oxide and tumor necrosis factor-α.

Analysis of the neutral polysaccharide fraction of MCP and its inhibitory activity on galectin-3

The pH-modified citrus pectin (MCP) has been demonstrated to inhibit galectin-3 in cancer progression. The components and structures of MCP related to this inhibition remained unknown. In this paper, we fractionated MCP on DEAE-cellulose column into a homogenous neutral fraction MCP-N (about 20kDa) and a pectin mixture fraction MCP-A (wide molecular distribution on Sepharose CL-6B chromatography). Both MCP-N and MCP-A inhibited hemagglutination mediated by galectin-3 with minimum inhibition concentration (MIC) 625 and 0.5μg/ml, respectively. MCP-N was identified to be a type I arabinogalactan (AG-I) with a main chain of β-1→4-galactan. MCP-N was digested by α-L-arabinofuranosidase to give its main chain structure fraction (M-galactan, around 18kDa), which was more active than the original molecule, MIC 50μg/ml. The acidic degradation of M-galactan increased the inhibitory activity, MIC about 5 times lower than M-galactan. These results above showed that the functional motif of the β-1→4-galactan fragment might lie in the terminal residues rather than in the internal region of the chain. Therefore, MCP-N and its degraded products might be developed to new potential galectin-3 inhibitors. This is the first report concerning the fractionation of MCP and its components on galectin-3 inhibition. The information provided in this paper is valuable for screening more active galectin-3 inhibitors from natural polysaccharides.

Isolation, chemical characterization and in vitro antioxidant activities of polysaccharides from Aconitum coreanum

The water-soluble crude polysaccharide, Aconitum coreanum polysaccharides (ACPS), was extracted from A. coreanum and purified by Diethylaminoethyl cellulose (DEAE-C) and Sepharose CL-6B chromatography, giving three polysaccharide fractions coded as ACPSA-1, ACPSB-2 and ACPSB-3. Their chemical and physical characteristics of polysaccharide fractions, and antioxidant capacity, including scavenging activity against DPPH radicals, superoxide and hydroxyl radicals and chelating ability were valuated in this paper. Experimental results showed that ACPS-2 and ACPS-3 exhibited significantly antioxidant activity at a concentration-dependent manner. The polysaccharide fractions can be developed as new antioxidant agents. Key words: Aconitum coreanum, polysaccharide, purification, characterization, antioxidant activity.