Polysaccharides of algae 68. Sulfated polysaccharides from the Kamchatka brown alga Laminaria bongardiana

Several fucoidan fractions were isolated from the biomass of the Kamchatka brown alga Laminaria bongardiana by hot water extraction followed by anion-exchange chromatography. Fucoidans were found to be composed of l-fucose, d-galactose, and sulfate as the major components, whereas d-xylose, d-mannose, d-glucuronic acid, and acetate were detected as the minor constituents. Highly sulfated fucoidan fractions F-2 and F-3 were solvolytically desulfated by heating in dimethyl sulfoxide in the presence of pyridine. The structures of native and desulfated polysaccharides were investigated by the methylation analysis and NMR spectroscopy. It was shown that F-2 contains fucan sulfate, the backbone of which is made of 1→3-linked α-l-fucopyranose residues with single α-l-Fucp branches at positions 2 and sulfate groups predominantly at positions 4. Sulfated fucoglucuronomannan, fucoglucuronan, and fucogalactan were detected in F-2 as concomitant polysaccharides. Fucan sulfate and sulfated fucogalactan were the major components of the fraction F-3. The anticoagulant properties of fucoidan fractions were assessed. It was demonstrated that the activity of the fraction F-3 is comparable with that of low-molecular-weight heparin (enoxaparin), whereas the activity of total fucoidan F and the fraction F-2 is ~2/3 and ~1/2, respectively, of the activity of F-3, which is in accordance with the lower sulfate content in these samples. Desulfated preparations F-2deS and F-3deS were completely devoid of anticoagulant activity.

     

Effects of Holdfast of Laminaria japonica on Listeria Invasion on Enterocyte-Like Caco-2 Cells and NO Production of Macrophage RAW 264.7 Cells

Listeria monocytogenes (Lm) causes food poisoning in humans mainly through consumption of ready-to-eat foods. Immunocompromised persons are at the highest risk for infection. We investigated effects of crude soluble polysaccharides (SPS) and ethanolic extract (EE) fractions of frond (kombu) and holdfast (ganiashi) parts of Laminaria japonica on Lm invasion into human enterocyte-like Caco-2 cells and immune and/or inflammatory reactions of murine macrophage RAW 264.7 cells. Recovery and viscosity were high in kombu SPS. Total phenolic content and antioxidant activities (2,2-diphenyl-1-picrylhydrazyl radical scavenging capacity and Fe-reducing power) were higher in ganiashi EE. EE of ganiashi, rather than kombu, suppressed the Lm invasion into the differentiated Caco-2 cells, though the inhibitory effect of SPS was not significant. Ganiashi SPS increased the nitric oxide (NO) production of intact RAW 264.7 cells. On the other hand, the NO production from Escherichia coli O111 lipopolysaccharide-activated cells was suppressed by kombu SPS and ganiashi EE. These results suggest that L. japonica, particularly ganiashi, might suppress the invasion and infection of Lm and also the inflammation.     

Chemical structure characterization of polysaccharide from Osmunda japonica Thunb and its inhibitory activity on uterine fibroids

The active component polysaccharide of Osmunda japonica Thunb was extracted and its chemical structure was detected by FT-IR, 1D-NMR and 2D-NMR spectra, in order to have a positive preventive and therapeutic effect on uterine fibroids. The physicochemical properties of Osmunda japonica Thunb polysaccharides (OPP) were analyzed, and the results showed that OPP was heteropolysaccharides composed of six monosaccharides. Ara and Glc were the main monosaccharides of the polysaccharide, with a molecular weight of 252.54 ± 10.84 kDa. The main chain was connected by 1,3-linked Araf, which had a high degree of branching, and was connected to the branch at the O-2 and O-3 positions. The mice model of uterine fibroids was established by the estrogen load method. The coefficient of the uterus in the mice model increased significantly, irregular nodular processes were observed in the uterus, focal hyperplasia appeared in the smooth muscle layer, cell enlargement, disordered arrangement of smooth muscle cells, and endometrial hyperplasia was obvious. Furthermore, this work found that the serum estradiol (E2) and progesterone (P) concentrations in the model group were significantly higher than those in the normal group (P < 0.01), indicating successful modeling. OPP decreased serum E2 and P levels in a dose-dependent manner. Collectively, OPP had an obvious inhibitory effect on uterine fibroids in estrogen-loaded mice.     

鱿鱼墨多糖的硫酸酯化及抗凝血活性

本文采用三氧化硫吡啶复合物二甲亚砜法首次对北太平洋鱿鱼墨多糖SIP进行硫酸酯化。对硫酸酯化后多糖样品的基本化学组成进行测定,分析了糖组成,硫酸基含量和分子量。并结合红外光谱和一维核磁分析其结构,结果表明硫酸酯化主要发生在GalNAc的4,6位上。进一步的凝血活性分析表明有较好的延长APTT和PT时间效果。对凝血因子的抑制实验则表明,硫酸化后的鱿鱼墨多糖TBA-1对FIIa和FXa 均有显著的抑制作用。     

Engineered bio-active polysaccharides from heparin

[Image: see text] Heparin, the well-known anticoagulant polysaccharide, is also active in many other biological systems owing to its structural similarity to HS, but usually lacks selectivity because it is more highly sulfated. A series of straightforward chemical reactions (de-O-sulfation, de-N-sulfation and re-N-acetylation), carried out to partial or complete extent, were combined, resulting in a number of modified heparin polysaccharide derivatives with altered properties. These exhibited a range of abilities to promote cell signalling through the FGF/FGFR tyrosine kinase signalling system, in an in vitro cell assay with combinations of FGF-1, -2, -3 and FGFR 1 and 3. One polysaccharide (N-acetylated, 6-O- and 2-O-sulfated heparin), with only a fraction (<10(-3)) of the anticoagulant activity of heparin (200 U . mg(-1)), promoted FGF-2-mediated angiogenesis (10-fold) and therefore had an improved ratio of pro-angiogenic activity to anticoagulant activity in excess of 10(4) compared to heparin. These results demonstrate that heparin-derived polysaccharides can be engineered for selected activities and have potential in a wide range of medical, biotechnological and tissue-engineering applications. Effect of selected engineered heparin polysaccharides on angiogenesis.     

Combinatorial enzymatic synthesis of heparan sulfate

Escherichia coli K5 heparosan was enzymatically modified by Chen and colleagues to construct a library of heparan sulfate polysaccharides for evaluation, leading to the discovery that a 2-O-sulfoiduronic acid residue is not essential for antithrombin-mediated anticoagulant activity in larger oligosaccharide and polysaccharide structures.     

Structural Characterization and In-Vitro Antioxidant and Immunomodulatory Activities of Polysaccharide Fractions Isolated from Artemisia annua L.

Arimisia annua L. is an important anticancer herb used in traditional Chinese medicine. The molecular basis underpinning the anticancer activity is complex and not fully understood, but the herbal polysaccharides, broadly recognised as having immunomodulatory, antioxidant and anticancer activities, are potential key active agents. To examine the functions of polysaccharides from A. annua, their immunomodulatory and antioxidant potentials were evaluated, as well as their structural characterization. The water-soluble polysaccharides (AAPs) were fractionated using size-exclusion chromatography to obtain three dominant fractions, AAP-1, AAP-2 and AAP-3, having molecular masses centered around 1684, 455 and 5.8kDa, respectively. The antioxidant potentials of the isolated polysaccharides were evaluated by measuring radical scavenging activities against DPPH^● (2,2-diphenyl-1-picrylhydrazyl radical), ABTS^●+ (2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid radical ion), and the OH^● (hydroxyl radical). AAP-1 displayed high antioxidant activities against these radicals, which were 68%, 73% and 78%, respectively. AAP-2 displayed lower scavenging activities than the other two fractions. Immunostimulatory activities of AAPs were measured using mouse macrophages. The three polysaccharide fractions displayed significant antioxidant activities and stimulated the production of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). AAP-1 showed significant immunostimulatory activity (16-fold increase in the production of IL-6 compared to the control and 13-fold increase in the production of TNF-α) with low toxicity (>60% cell viability at 125 μg/mL concentration). Preliminary structural characterization of the AAPs was carried out using gas chromatography (GC) and FTIR techniques. The results indicate that AAP-1 and AAP-2 are pyranose-containing polysaccharides with β-linkages, and AAP-3 is a β-fructofuranoside. The results suggest that these polysaccharides are potential candidates for immunotherapy and cancer treatment.     

Isolation,Characterization, and Medicinal Potential of Polysaccharides of Morchella esculenta

Mushroom polysaccharides are active medicinal compounds that possess immune-modulatory and anticancer properties. Currently, the mushroom polysaccharides krestin, lentinan, and polysaccharopeptides are used as anticancer drugs. They are an unexplored source of natural products with huge potential in both the medicinal and nutraceutical industries. The northern parts of Pakistan have a rich biodiversity of mushrooms that grow during different seasons of the year. Here we selected an edible Morchella esculenta (true morels) of the Ascomycota group for polysaccharide isolation and characterization. Polysaccharopeptides and polysaccharides from this mushroom were isolated using the green chemistry, hot water treatment method. Fourier transform infrared spectroscopy revealed the sugar nature and possible beta-glucan type structure of these polysaccharides. Antioxidant assays showed that the deproteinized polysaccharides have moderate free radical scavenging activity. These isolated polysaccharides exhibited good acetylcholinesterase (AChE) and butyryl cholinesterase (BChE) inhibition activities. Therefore, these polysaccharides may be valuable for the treatment of Alzheimer’s and Parkinson’s diseases. Further bioassays are needed to discover the true potential of M. esculenta polysaccharides for medicinal purposes.     

Chemical evaluation of <Emphasis Type="Italic">Fallopia</Emphasis> species leaves and antioxidant properties of their non-cellulosic polysaccharides

Utilization of biomass from forest or agricultural crops for the production of energy or chemical products provides environmental advantages. Leaves of the knotweeds Fallopia japonica, Fallopia sachalinensis, and Fallopia × bohemica are rich sources of phenolics and polysaccharides. In view of their potential utilization before the disposal of these invasive plants, their gross composition was investigated. The content of extractives was the highest in F. sachalinensis. Extractive-free leaves of all species showed similar chemical composition. The content of ash, protein, lignin, uronic acids, and α-cellulose in the leaves of F. sachalinensis, F. × bohemica, and F. japonica was 63.3 %, 64.1 %, and 63.4 %, respectively. The rest comprised hemicelluloses and neutral carbohydrate components of pectic polysaccharides. Sequential extraction of F. sachalinensis with water, EDTA, DMSO, 1 % NaOH, and 5 % NaOH yielded fractions accounting together for 27.6 % of polysaccharides. Pectic polysaccharides predominated in the first three fractions, whereas the hemicellulose components — xylan and xyloglucan, prevailed in the two alkaline fractions. The polysaccharides displayed significant radical scavenging activities in the 1,1-diphenyl-2-picrylhydrazyl free radical assay thus indicating their potential application as novel natural antioxidants.     

Hepatoprotective Polysaccharides from Geranium wilfordii: Purification,Structural Characterization,and Their Mechanism

Traditional Chinese Medicine is generally used as a decoction to guard health. Many active ingredients in the decoction are chemical ingredients that are not usually paid attention to in phytochemical research, such as polysaccharides, etc. Based on research interest in Chinese herbal decoction, crude polysaccharides from G. wilfordii (GCP) were purified to obtain two relatively homogeneous polysaccharides, a neutral polysaccharide (GNP), and an acid polysaccharide (GAP) by various chromatographic separation methods, which were initially characterized by GC-MS, NMR, IR, and methylation analysis. Studies on the hepatoprotective activity of GCP in vivo showed that GCP might be a potential agent for the prevention and treatment of acute liver injury by inhibiting the secretion levels of ALT, AST, IL-6, IL-1β, TNF-α, and MDA expression levels, increasing SOD, and the GSH-Px activity value. Further, in vitro assays, GNP and GAP, decrease the inflammatory response by inhibiting the secretion of IL-6 and TNF-α, involved in the STAT1/T-bet signaling pathway.     

Synthesis of Sulfated Polysaccharides and Oligosaccharide Derivatives with Potent Anti-Aids Virus Activity

Abstract

Sulfated synthetic polysaccharides (with both high anti-AIDS virus activity and high anticoagulant activity) were prepared by sulfating such synthetic polysaccharides as ribopyranan, ribofuranans, and dextrans. Sulfated natural polysaccharides with high anti-AIDS virus activity but low anticoagulant activity were synthesized from lentinan and curdlan. It is assumed that curdlan sulfate will be helpful as an AIDS drug. In addition, sulfated alkyl oligosaccharides with high anti-AIDS virus activity were prepared.     

Structural properties and antioxidant activities of polysaccharides isolated from sunflower meal after oil extraction

Making full use of sunflower seeds, including oil and the polysaccharides extracted from the meals which oil has been extracted, is one way to enhance their industrial value. Such meals contain abundant polysaccharides; however, the application of polysaccharides isolated from sunflower remaining meals after oil extraction has not been investigated. In this study, polysaccharides were isolated by alkali from sunflower meals after different oil extraction processes, and their structural properties and antioxidant activities were compared. The results indicated that these polysaccharides displayed significant variability in monosaccharide composition and molecular weight. Differences in structural properties could result in differences in functional antioxidant properties. The polysaccharide (SPHE-1) obtained from the meals after traditional hexane extraction exhibited the best antioxidant activities, including DPPH free radical-scavenging assay and hydroxyl radical scavenging activity among all the polysaccharide fractions. The research provides valuable information for making efficient use of sunflower seeds in the food industry.     

水溶性高支化多糖及硫酸酯衍生物抗癌活性研究

从虎奶菇菌核中提取出一种水溶性高支化葡聚糖,并用氯磺酸和吡啶在无水二甲亚砜中进行硫酸酯衍生化,制备出硫酸酯衍生物.用巴比赛小鼠动物实验研究该多糖及其硫酸酯衍生物的体内抗肉瘤S-180活性,以及用肝癌细胞株HepG2研究体外活性,同时讨论多糖分子尺寸与抗癌活性的关系.通过形态学、组织切片、以及酶联免疫法研究硫酸酯化对多糖抗肿瘤活性的影响.结果表明,支链多糖的均方根旋转半径在42.5~113 nm之间,显示出较高的体内和体外抗肿瘤活性,硫酸酯衍生物的均方根旋转半径在17.8~34.9 nm之间,抗癌活性随其分子尺寸的增大而逐渐提高.多糖及其硫酸酯衍生物能诱导人体肝癌HepG2细胞凋亡,且具有时间依赖性.它们对免疫反应的调节作用可通过肿瘤坏死因子TNF-α来介导.     

Catalytic Properties of Mechanically Activated Humic Substances in Electroreduction of Oxygen

Effect of mechanical treatment of peat on the yield of the major fractions (polysaccharides, poly- phenols, water-soluble compounds, and humic acids) and catalytic activity of humic substances in oxidation processes is studied. Conditions of formation of substances with the maximal antioxidant and initiating activities are determined. The antioxidant and initiating activities of the polysaccharide and humic fractions is studied, in relation to the structure and iron content.     

Characterization and Antioxidant Activity of Mannans from Saccharomyces cerevisiae with Different Molecular Weight

Polysaccharides were extracted from natural sources with various biological activities, which are strongly influenced by their chemical structure and molecular weight. In this research, mannans polysaccharides were obtained from Saccharomyces cerevisiae by ethanol precipitation. The molecular weight of YM50, YM70, and YM90 mannans was 172.90 kDa, 87.09 kDa, and 54.05 kDa, respectively. Scanning electron microscopy of YM 90 mannans showed a rough surface with numerous cavities, while the surfaces of YM50 and YM70 were relatively smooth. Sepharose CL-6B and FTIR indicated that mannans had the characteristic bands of polysaccharides. The antioxidant activities of polysaccharides were evaluated in vitro using various assays. Mannans showed a good scavenging activity of DPPH radicals which depend on the molecular weight and concentration, and a higher scavenging activity of hydroxyl radical than ferric-reducing power activities. For the three types of mannans, cytotoxicity and hemolytic activity were rarely detected in mice erythrocytes and Caco-2 cells. Those results could contribute to the further application of mannans from Saccharomyces cerevisiae in the food and medicine industry.     

Isolation and Characterization of Low Molecular Weight Glycosaminoglycans from Marine Mollusc Amussium pleuronectus (Linne) using Chromatography

The glycosaminoglycan (GAG) heparin is a polyanionic sulfated polysaccharide most recognized for its anticoagulant activity. In the present study, the GAGs were extracted from bivalve mollusc Amussium pleuronectus. The crude GAGs were fractionated by ion-exchange (DEAE-cellulose and Amberlite IRA-900 & 120) chromatography. The recovered active fractions (as determined by metachromatic assay) were confirmed by agarose gel electrophoresis and the active fractions were purified in Sephadex G-100 column. Fractionated and purified GAG molecular weight was determined through gradient polyacrylamide gel electrophoresis. The structural characterization of low molecular weight GAG was analyzed by Fourier transform infrared spectroscopy. The activated partial thromboplastin time of purified GAG is 95 IU/mg and has molecular weight 6,500–7,500 Da. The disaccharide compositional analysis on the GAG sample was sulfated like porcine intestinal mucosal heparan sulfate, and it contains equivalent amount of uronic acid and hexosamine. The results of this study suggest that the GAG from A. pleuronectus could be an alternative source of heparin.     

Effects of a High-Molecular-Weight Polysaccharides Isolated from Korean Persimmon on the Antioxidant,Anti-Inflammatory,and Antiwrinkle Activity

Persimmon (Diospyros kaki), a familiar and widespread fruit worldwide, is known to exhibit several physiological effects because of the presence of pharmacologically active compounds called phytochemicals. However, its high-molecular-weight compounds, particularly polysaccharides, have not been extensively studied. In this study, D. kaki extract (DK) was fractionated into low- and high-molecular-weight fractions (DK-L and DK-H, respectively) through ethanol fractionation, and their effects on antioxidant, anti-inflammatory, and antiwrinkle activities were investigated by an in vitro system. DK-H contained significantly higher contents of neutral sugar, uronic acid, and polyphenols compared to DK and DK-L. Furthermore, DK-H exhibited significantly improved pharmacological activities, such as antioxidant, anti-inflammatory, and antiwrinkle properties, compared to those of DK and DK-L, demonstrating that DK-H may play an important role in mediating the beneficial effects of persimmon. Sugar composition analysis and molecular characterization indicated that DK-H consisted of a galacturonic acid (GalA)-rich polysaccharide with a molecular weight of >345 kDa that mainly comprised GalA and small amounts of neutral sugar and polyphenol residues. These results suggest that the bioactive fraction DK-H is likely to be a GalA-rich pectic polysaccharide containing a small number of polyphenol residues, which may be a novel candidate in the pharmaceutical and cosmeceutical industries.     

Antiviral activity of polysaccharides against infection of tobacco mosaic virus

In the course of searching for antiviral substances to tobacco mosaic virus (TMV), it was found that polysaccharides have a high inhibitory activity against TMV infection. The leaves of Xanthi NN tobacco were rubbed with the mixtures of TMV and polysaccharides such as chondroitin sulfate C- and A- types. The addition of polysaccharides to the inoculum solution greatly reduced the number of local lesions formed on the inoculated leaves. Here the polysaccharide did not completely prevent virus entry into the leaves and the virus particles may penetrate and multiply in leaves without forming lesions. Although the electron micrograph showed that the virus suspension was almost monodisperse, the addition of polysaccharides caused TMV to form large raft-like aggregates. The TMV solution became turbid after the addition of a large amount of polysaccharides. A threshold concentration of polysaccharides exists for virus precipitation, which is independent of the virus concentration. The size of polysaccharide at the threshold concentration agreed well with that obtained by light scattering method. The strength of the interaction between TMV and polysaccharides was found to be related to the degree of inhibitory activity of polysaccharides.     

杜氏盐藻多糖的高效体积排阻色谱的保留特性及其分析方法的研究

通过对从杜氏盐藻中提取出的不同多糖级分在高效体积排阻色谱柱(Waters Ultrahydragel Linear,7.8 mm i.d.×300 mm,2根串联)上的保留特性的考察及其分离分析条件的优化,建立了高效体积排阻色谱分析盐藻多糖平均相对分子质量及其分布的方法。结果表明:流动相中盐的种类及其浓度、pH值对3种酸性多糖级分(特别是硫酸化多糖级分PD4a)的保留行为有显著影响;在柱温为45 ℃,流速为0.9 mL/min条件下,使用0.1 mol/L的NaAc水溶液作流动相基本上能消除非特异性吸附作用及分子间缔合等因素的干扰,使各多糖级分基本以非缔合状态按立体排除机制保留和分离。在优化的色谱条件下,测得的盐藻多糖5个级分的重均相对分子质量(Mw)分别为1548000,33000,67000,424000,10000;测得的硫酸化多糖级分PD4a的Mw和峰面积的相对标准偏差分别为1.7%和 0.88%(n=5)。