Water-soluble polysaccharides from Angelica sinensis (Oliv.) Diels: Preparation, characterization and bioactivity

Crude water-soluble polysaccharides (ASP) were separated from Angelica sinensis (Oliv.) Diels by hot water extraction. They were fractionated into neutral and acidic polysaccharides by anion-exchange chromatography. The neutral polysaccharide (ASP1) was rich in glucose, galactose, and arabinose suggesting a mixture of glucan and arabinogalactan. The acidic polysaccharide (ASP2, ASP3) consisted mainly of galacturonic acid along with rhamnose, arabinose, and galactose indicating a pectic polysaccharide. The degree of esterification of ASP and ASP3 were 54.06% and 47.14% for the crude and purified sample, respectively. ASP3, with a molecular weight of 3.4 × 10⁴ Da determined by high-performance size-exclusion chromatography (HPSEC), was the major constituent for the crude extracts. The radioprotective effect of the pectic polysaccharide ASP3 was studied in murine models. ASP3 pretreated mice exhibited a significant decrease of apoptosis (P < 0.05, dosage of 200 mg/kg d body weight) in peripheral lymphocytes compared to the irradiated control. The results showed that ASP3 can protect leucocytes and lymphocytes of mice against radiation induced damage, which has potential radioprotective effect on acute radiation injured mice.     

In vitro anti-herpetic activity of sulfated polysaccharide fractions from Caulerpa racemosa

A sulfated polysaccharide fraction was isolated from the hot water extract of the green alga Caulerpa racemosa and designated HWE. This polymer, which contained galactose, glucose, arabinose and xylose as the major component sugars, had [alpha](D)(30) + 46.2 degrees in water and contained 9% sulfate hemiester groups. Sugar linkage analysis indicates that HWE was branched and mainly contained 1,3- and 1,3,6-linked galactose, 1,3,4-linked arabinose, 1,4-linked glucose and terminal- and 1,4-linked xylose residues. Sulfation was deduced from infrared spectroscopy and methylation analysis to occur on O-6 of galactose and O-3 of arabinose. The native polysaccharide could be fractionated by size exclusion chromatography into two overlapping fractions and the major fraction has a hydrodynamic volume similar to that of 70 kDa dextran. HWE was a selective inhibitor of reference strains and TK(-) acyclovir-resistant strains of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in Vero cells, with antiviral effective concentration 50% (EC(50)) values in the range of 2.2-4.2 microg/ml and lacking cytotoxic effects. Furthermore, HWE did not exhibit anticoagulant properties at concentrations near the EC(50).     

Structure of Hemicellulose Isolated from Rice Endosperm Cell Wall : Mode of Linkages and Sequences in Xyloglucan, β-Glucan and Arabinoxylan

A hemicellulosic polysaccharide, which was homogeneous on sedimentation analysis and also on electrophoresis, was isolated from the rice endosperm cell walls by the combination of alkaline extraction, ion exchange chromatography and iodine complex formation. It is composed of arabinose, xylose and glucose (molar ratio, 1.0: 2.0: 5.7) together with a small amount of galactose and rhamnose. Methylation analysis, Smith degradation and fragmentation with cellulase showed that this polysaccharide is composed of three distinct polysaccharide moieties i.e., xyloglucan, β-glucan and arabinoxylan. The xyloglucan consists of β-(1→4)-linked glucan back bone and short side chains of single xylose units or galactosylxylose both attached to C-6 of the glucose residues. The β-glucan contains both (1 →3)-and (1→4)-linkages similarly to the other cereal β-glucans, but differ from them in containing the blocks of (1→3)-linked glucose residues in the chain. The arabinoxylan has a highly branched structure, in which 78% of (1→4)-linked xylose residues have short side chains of arabinose at C-3 position.

On the basis of these findings, the interconnection of these polysaccharide moieties is discussed.     

Studies on the Carbohydrates of Horse Radish,Cochlearis Armoracia L

In the root of horse radish, sucrose, fructose, glucose and two unknown fructose-oligosac-charides were found in the form of free sugars, in the decreasing order just given, and large amounts of starch, hot 50% methanol-soluble polysaccharide, water-soluble polysaccharide, pectin, hemicellulose, cellulose and a small amount of lignin were found as polysaccharides. As the constructive sugar of these polysaccharides, the following, i.e., glucose, fructose, galactose, arabinose, galacturonic acid and xylose were detected.     

Characterization and antioxidant activity of Ginkgo biloba exocarp polysaccharides

Ginkgo biloba exocarp polysaccharide (GBEP) was obtained by hot water extraction, the crude polysaccharide was deproteinized by Sevag method and fractionized by a DEAE Sepharose fast flow anion-exchange column. Five fragments were obtained, including neutral polysaccharide (GBEP-N) and four acidic polysaccharides (GBEP-A1, GBEP-A2, GBEP-A3 and GBEP-A4). GBEP-N and GBEP-A3 were further purified by Superdex 200 gel column chromatography. The resulted two fractions GBEP-NN, and GBEP-AA were characterized by FT-IR, and HPGFC (high pressure gel filtration chromatography). Monosaccharide composition was determined by RP-HPLC method of precolumn derivatization with 1-phenyl-3-5-pyrazolone. GBEP-NN was mainly composed of rhamnose, arabinose, mannose, glucose and galactose, while GBEP-AA was mainly made up of mannose, rhamnose, glucuronic acid, galacturonic acid, galactosamine, glucose, galactose, xylose, arabinose, and fucose. The crude GBEP exhibited certain antioxidant activity. At the concentration of 5 mg/mL, the hydroxyl radical scavenging effect of GBEP was 90.52%, greater than 77.37% for the positive control ascorbic acid.     

Extraction, partial characterization and bioactivity of polysaccharides from boat-fruited sterculia seeds

Three polysaccharides (water-soluble (WSP), alkali-soluble (ASP) and insoluble (IMP)) from boat-fruited sterculia seeds were obtained using different extraction methods. Moisture, ash, protein and total carbohydrate content of WSP, ASP and IMP were analyzed. WSP was rich in glucose, rhamnose, arabinose and galactose while small amount of xylose was also detected. The monosaccharide composition as well its relative content for WSP and ASP were similar. The intrinsic viscosity results demonstrated that ASP had much lower intrinsic viscosity than WSP, indicating partial polysaccharides were degraded into low molecular weight polymers during alkaline extraction. The acute anti-inflammatory bioactive results of polysaccharides indicated that WSP demonstrated an inhibitive effect toward acute inflammation.     

Studies of the polysaccharides from sunflower heads

Extraction of sunflower heads with ammonium oxalate afforded water-soluble pectin material and water-insoluble glycoprotein material, the carbohydrate portion of which consisted of galacturonic acid and xylose residues; the pectin material defied fractionation with cetylpyridinium chloride. Extraction with hydrochloric acid (pH 1.5) afforded water-soluble and water-insoluble polysaccharide materials. The former, when fractionated with cetylpyridinium chloride, gave a glycoprotein, the carbohydrate moiety of which was composed of galacturonic acid, galactose (major), glucose, arabinose, and xylose, and also a rhamnan. The latter was a glycoprotein, the carbohydrate portion of which consisted of galactose (major), glucose, xylose, and rhamnose residues. Extraction of the sunflower heads with water also gave glycoprotein material, which was fractionated by paper electrophoresis into a glyco-protein, the carbohydrate moiety ofwhich was composed of galacturonic acid (minor), galactose, glucose, xylose, arabinose, and rhamnose (major) residues, and a heteropolysaccharide composed of galactose (major), glucose, xylose, and arabinose residues.     

Stimulation of polysaccharide formation by 2,4-dichlorophenoxyacetic acid in callus tissues of Arabidopsis thaliana

A relatively high concentration of 2,4-dichlorophenoxyacetic acid (45 μ M ) in solid culture medium stimulated the formation and secretion of mucilage polysaccharides by callus tissues of Arabidopsis thaliana L. Heynh. (line Estland). The mucilage was composed of at least two polysaccharides as revealed by gel chromatography on Sepharose 4B: the major component (87%) eluted in the void volume (molecular weight 2 × 10⁶ or greater) and the minor component (13%) eluted in the molecular weight range from 2 × 10⁴ to 4 × 10⁵. Both polysaccharide components contained small amounts of uronic acids. The major polysaccharide consisted mostly of galactose (49%), arabinose (28%) and fucose (10%), whereas the minor one consisted of galactose (44%), xylose (18%), arabinose (14%) and rhamnose (14%). One of the components of the secreted mucilage seems to be an arabinogalactan.     

An arabinogalactan from the skin of Opuntia ficus-indica prickly pear fruits

The cold-water extract from the skin of Opuntia ficus-indica fruits was fractionated by anion-exchange chromatography. The major fraction, which was purified by size exclusion chromatography, consisted of a polysaccharide composed of galactose and arabinose residues in the ratio 6.3:3.3, with traces of rhamnose, xylose and glucose, but no uronic acid. The results of methylation analysis, supported by (13)C NMR spectroscopy, indicated that this polysaccharide corresponded to an arabinogalactan having a backbone of (1-->4)-linked beta-D-galactopyranosyl residues with 39.5% of these units branched at O-3. The side-groups consisted either of single L-arabinofuranosyl units or L-arabinofuranosyl alpha-(1-->5)-linked disaccharides. This polysaccharide is thus an arabinogalactan that can be classified in the type I of the arabinogalactan family.     

Patterns of polysaccharide biosynthesis in differentiating cells of maize root-tips

1. The patterns of incorporation of radioactivity from d-[6-(3)H]-, d-[1-(14)C]-, d-[U-(14)C]- and d-[6-(14)C]-glucose and [U-(14)C]myoinositol into the neutral sugars and uronic acids of the polysaccharides synthesized in different regions of the root-tip of maize were determined. 2. The root-cap tissue synthesized a slime in which a polysaccharide that contained a high proportion of fucose (32%) and galactose (21%) was found. This polysaccharide is synthesized only by the root-cap cells, and very little polysaccharide containing fucose is synthesized in adjacent tissues. Part of the meristematic tissue of the root is surrounded by the cap cells. A section of the root that contains both these tissues can be analysed, and the polysaccharide synthesized by the meristematic region can be obtained since the contribution of the root-cap cells can be found by the amount of fucose formed. 3. It was shown that there is very little difference in the polysaccharide synthesis of the meristematic region from that of the cells immediately behind it. In the more mature cells, however, the amount of xylose synthesized relative to that of arabinose is increased, and the proportion of xylose and arabinose formed in the matrix polysaccharides is increased whereas that of galactose is decreased. 4. The effect of 2,4-dichlorophenoxyacetic acid (2,4-D) on polysaccharide synthesis was to bring about a decrease in the relative amount of galactose synthesized in the matrix polysaccharides of cells immediately adjacent to the meristematic region and also in the more mature tissue. The growth factor also increased the amount of xylose synthesized relative to that of arabinose in the more mature tissue. These metabolic effects were related to a very obvious change in the morphological appearance of the root-tips. 5. Radioactivity from [U-(14)C]myoinositol was incorporated mainly into xylose, arabinose and galacturonic acid rather than into the hexoses, although small amounts of these sugars were formed.     

Composition analysis and antioxidant activity of polysaccharide from Dendrobium denneanum

Three polysaccharide fractions (DDP1-1, DDP2-1 and DDP3-1) were successfully purified from the crude polysaccharide of Dendrobium denneanum by DEAE-Cellulose and Sephadex G-200 column chromatography. The average molecular weights (Mws) of these fractions were 51.5, 26.1 and 6.95 kDa, respectively. Monosaccharide components analysis indicated that DDP1-1 and DDP2-1 were composed of arabinose, xylose, mannose, glucose and galactose in a molar ratio of 1.00:2.82:57.11:140.82:7.76 and 1.00:1.62:1.18:77.5:7.79. DDP3-1 was composed of arabinose, mannose, glucose and galactose in a molar ratio of 1.00:1.03:8.84:2.00. On the basis of antioxidant test in vitro, DDP2-1 exhibited the highest antioxidant ability among these samples.     

Isolation and characterization of polysaccharides from Tansy

Using extraction with 0.75% aqueous ammonium oxalate, the following polysaccharide fractions were isolated: tanacetans TVF, TVS, and TVR from floscules, sprouts, and roots, respectively, of Tanacetum vulgare L., spread throughout the European North of Russia. The sugar chain of tanacetan TVF consists of D-galacturonic acid (61.4%), arabinose (14.7%), galactose (10.2%), and rhamnose (3.7%) as the main constituents as well as xylose, glucose, mannose, apiose, and 2-O-methylxylose in trace amounts. Tanacetans TVS and TVR were shown to differ in the sugar quantitative composition. They contain 67 and 28% galacturonic acid, respectively. A partial acid hydrolysis of the tanacetan TVF gave a polysaccharide fragment TVF1, alpha-1,4-D-galacturonan (GalA 98.2%). Digestion with pectinase (alpha-1,4-D-polygalacturonase) resulted in fragment TVF3, containing residues of arabinose (27.1%) and galactose (17.3%). NMR spectroscopy allowed detection of the terminal residues of alpha-Araf and beta-Galp as well as of the residues of alpha-Araf substituted in 3,5- and 5-positions. Thus, tanacetan TVF was proved to be a pectic polysaccharide.     

Changes in cell-wall polysaccharides in relation to seedling development and the mobilisation of reserves in the cotyledons of Lupinus angustifolius cv. Unicrop

Some 22% of the dry weight of the cotyledons of resting seeds of Lupinus angustifolius cv. Unicrop has been shown to be non-starch polysaccharide material comprising the massively thickened walls of the storage mesophyll cells. On hydrolysis this material released galactose (76%), arabinose (13%), xylose (4%), uronic acid (7%): only traces of glucose were detected indicating the virtual absence of cellulose from the walls. Changes in the amount and composition of this material following germination have been studied in relation to parameters of seedling development and the mobilisation of protein, lipid and oligosaccharide reserves. Starch, which was not present in the resting seed, appeared transitorily following germination: under conditions of continuous darkness starch levels were reduced. During the period of bulk-reserve mobilisation, 92% of the non-starch polysaccharide material disappeared from the cotyledons. The residual cell-wall material released galactose (14%), arabinose (19%), xylose (24%) and uronic acid (43%). The galactose and arabinose residues of the cotyledonary cell walls clearly constitute a major storage material, quantitatively as important as protein. The overall role of the wall polysaccharides in seedling development is discussed.     

猴头菌子实体和菌丝体多糖的分离纯化与理化特征的比较*

人工栽培的猴头菌子实体和固体培养的菌丝体分别经热水提取,浓缩、醇析后得子实体粗多糖（HFP）和菌丝体粗多糖（HMP）,HFP的得率和多糖含量均高于HMP;两者再经透析、Sevag法脱蛋白、乙醇分级沉淀、SephadexG-100柱层析纯化,得子实体纯多糖hfp-1和菌丝体纯多糖 hmp-2,经HPLC检测hfp-1和hmp-2为单一均匀多糖。气相色谱分析表明hfp-1的单糖组成为阿拉伯糖、甘露糖、半乳糖和葡萄糖,摩尔比为：0.12：0.04：1.00：0.71;hmp-2的单糖组成为阿拉伯糖、木糖、甘露糖、半乳糖和葡萄糖,摩尔比为：0.25：0.41：0.31：1.00：0.29。紫外光谱分析表明组成中不含核酸和蛋白质,红外光谱分析二者均有多糖特征吸收峰,hfp-1有β-型连接的吸收峰,hmp-2无明显的β-型连接的吸收峰;经HPLC进行分子量测定,hfp-1的分子量为54000,hmp-2的分子量为68000。猴头菌子实体多糖和菌丝体多糖在化学组成上有一定的差异。     

Degradation of Perennial Ryegrass Leaf and Stem Cell Walls by the Anaerobic Fungus Neocallimastix sp. Strain CS3b

The degradation of cell walls isolated from stems and leaves of perennial ryegrass by the anaerobic fungus Neocallimastix sp. strain CS3b was studied in a defined medium. The combined cellulose and hemicellulose fraction represented 53.1 (wt/wt) and 63.3% (wt/wt) of the dry weight of control grass leaf and stem cell walls, respectively. In both leaf and stem cell walls, glucose was the major neutral monosaccharide, followed by xylose, arabinose, and galactose. After 2 days of fermentation with Neocallimastix sp. strain CS3b, treated cell walls contained smaller amounts of neutral sugars compared with those of undigested cell walls. These results were more evident for glucose, xylose, and arabinose than for galactose. Furthermore, the sugar content of leaf cell walls decreased before a decline in the sugar content of stem cell walls was observed. Data from formate and hydrogen production indicated that the growth of Neocallimastix sp. strain CS3b was completed in 4 days in the culture system used. During this period, the fungus liberated about 95% of the fermentable sugars in untreated material. On a percentage basis, no significant differences were found in final extent of degradation of glucose, xylose, and arabinose. Galactose, however, was degraded to a lesser extent.     

Anti-tumor and immunomodulating activities of a polysaccharide from the root of Sanguisorba officinalis L

A water-soluble polysaccharide, named as SOWP, was extracted and fractioned from the roots of Sanguisorba officinalis L. by DEAE-Sepharose anion exchange and Sepharose 6 Fast Flow column chromatography. The monosaccharide composition of SOWP determined by gas chromatography (GC) identified it was composed of glucose (68.5%), arabinose (13.2%), rhamnose (8.9), xylose (6.2) and galactose (3.0%). At the dose of 50, 100, and 200mg/kg, SOWP not only significantly inhibited the growth of mouse transplantable tumor, but also remarkably increased the spleen index and promoted splenocytes proliferation, macrophage phagocytosis and the production of serum cytokines IL-2, TNF-α and IFN-γ in Sarcoma 180-bearing mice. However, no direct cytotoxic activity against Sarcoma 180 cells was observed. The anti-tumor activity of the polysaccharide from S. officinalis maybe achieved by not directly cytotoxicity but rather immunopotentiation.     

Enzymic analysis of cell wall structure in apple fruit cortical tissue

Galactanase from Phytophthora infestans and an arabinosidase isoenzyme from Sclerotinia fructigena attacked the cortical cell walls of apple fruits liberating galactose and arabinose residues, respectively. Other arabinosidase isoenzymes from S. fructigena attacked cell walls very slowly. A S. fructigena polygalacturonase isoenzyme liberated half of the uronic acid residues with few associated neutral residues, while a second polygalacturonase isoenzyme released more uronic acid with a substantial proportion of arabinose and galactose and lesser amounts of xylose, rhamnose and glucose; reaction products of this enzyme could be further degraded by the first isoenzyme to give high MW fragments, rich in arabinose with most of the xylose, rhamnose and glucose, and low MW fragments rich in galactose and uronic acid. Endoglucanase from Trichoderma viride released a small proportion of the glucose residues from cell walls together with uronic acid, arabinose, xylose and galactose; more extensive degradation occurred if walls were pre-treated with the second polygalacturonase isoenzyme. Endoglucanase reaction products were separated into a high MW fraction, rich in arabinose, and lower MW fractions rich in galactose and glucose residues. The high MW polygalacturonase and endoglucanase products could be degraded with an arabinosidase isoenzyme to release about 75% of their arabinose. Cell walls from ripe fruit showed similar susceptibility to arabinosidase and galactanase to those from unripe apples. Cell walls from fruit, ripened detached from the tree were more susceptible to degradation by polygalacturonase than walls from unripe fruit or fruit ripened on the tree. Endoglucanase released less carbohydrate from ripe fruit cell walls than from unripe fruit cell walls.     

Purification and characterization of acidic proteo-heteroglycan from the fruiting body of Phellinus linteus (Berk. & M.A. Curtis) Teng

Phellinus linteus is a well-known oriental medicinal fungus that has various biological activities such as stimulation of humoral and cell mediated immunity, anti-mutagenicity activity, and anti-cancer activity. The process of isolating and purifying a water-soluble glycan from P. linteus was achieved by hot water extraction, filtration, solvent precipitation, dialysis, and freeze-drying. Acidic fractions of the polysaccharide were separated from crude polysaccharides by DEAE-cellulose anion exchange chromatography at 0.4 M NaCl. The molecular weight of the proteo-heteroglycan after Sepharose CL-4B gel filtration chromatography was about 150,000. The acidic proteo-heteroglycan consisted of 72.2% polysaccharide and 22.3% protein. The sugar of the proteo-heteroglycan was composed of mannose, galactose, glucose, arabinose, and xylose. The amino acid pattern showed that the fractions contained large amounts of aspartic acid, glutamic acid, alanine, glycine, and serine. The fractions for both alpha-glycan at 860 cm(-1) and beta-glycan at 910 cm(-1) had the characteristics of IR spectrum absorption as compared to those for beta-glucan derived from Lentinus edodes. A 13C and 1H NMR spectroscopy showed that the acidic proteo-heteroglycan was a noble biomolecule mixed both alpha- and beta-linkages, and a (1,6) branched type (1,3) glycan.     

Digestibility and energy value of non-starch polysaccharides in young chickens, ducks and geese, fed diets containing high amounts of barley

The purpose of the present investigation is to compare the ability of chickens, ducks and geese to digest and utilise a diet containing a relatively large amount of barley (40%) rich in beta-glucan (18 g kg(-1)) and NSP (137 g kg(-1)) of which 35 g kg(-1) were soluble non-starch polysaccharides (NSP). The diets were offered to the birds (50 chickens, 40 ducks and 30 geese) in the period from hatching to 42 days of age. The digestibility of NSP was measured during the last week of the growth period using chromic oxide as an indigestible marker. Emphasis was on total NSP, soluble and insoluble NSP and their constituent sugar residues (rhamnose, fucose, arabinose, xylose, mannose, galactose, glucose and glucuronic acid). The degradation of NSP to short chain fatty acids (SCFA) was determined in the small intestine, caeca and large intestine. Although significant differences were found between species to the extent of degradation of individual soluble, insoluble and total NSP residues in the small intestine and caeca, the overall apparent digestibility of total NSP was similar (39-42%). On the basis of the digestibility of the NSP sugar residues and the formation of SCFA in the gut, the energy value of NSP was estimated on 2.8, 3.2 and 2.7 kJ g(-1) NSP ingested (P>0.05) in chickens, ducks and geese, respectively. On average, NSP contributed approximately 3.5% of metabolisable energy (ME) in the three poultry species.     

A simple procedure for the synthesis of alpha-32P-nucleoside triphosphates.

Chemical analysis of grapefruit (Citrus paradisi) pectic polysaccharides demonstrated that galacturonic acid constitutes 78% by weight of the total carbohydrates found. The remaining 22% was accounted for by a number of sugars which include galactose, glucose, arabinose, xylose, and mannose and, by weight, galactose accounted for almost 50% of the total neutral sugar components found in these pectic polysaccharides. Treatment of pectic polysaccharides with galactose oxidase followed by reduction of oxidized galactose residues with tritiated potassium borohydride resulted in the labeling of pectic polysaccharides. Analysis of the labeled polysaccharides demonstrated that of the total radioactivity incorporated more than 90% was recovered in the galactose residues. These results clearly demonstrate the successful utilization of the galactose oxidase/tritiated potassium borohydride method in labeling plant pectic polysaccharide.