Homogeneous Polysaccharide Research Articles

Structural characterization of a polysaccharide from the flower buds of Tussilago farfara, and its effect on proliferation and apoptosis of A549 human non-small lung cancer cell line

In present study, we purified a polysaccharide, TFPB1, from the flower buds of Tussilago farfara using DEAE-cellulose 52 anion-exchange and Sephacryl S-300 HR gel filtration chromatography. TFPB1 was a homogeneous polysaccharide with a molecular weight of 37.8kDa and composed of rhamnose, galacturonic acid, glucose, galactose, and arabinose, in a ratio of 13:13:1:7:12. Methylation and NMR results demonstrated that TFPB1 contained a rhamnogalacturonan I backbone consisting of a repeat disaccharide unit →4)-α-D-GalAp-(1→2)-α-L-Rhap-(1→, substituted by various type II arabinogalactan branches including terminal galactose, (1→3)-β-D-galactan and (1→5)-α-L-arabinan, attached to the O-4 of (1→2)-α-L-Rhap. TFPB1 was found to inhibit cell proliferation of A549 cells and induce cell apoptosis in vitro. Furthermore, TFPB1 downregulated the phosphorylation of Akt, and upregulated caspase-3, Fas, FasL, and Bax expression, but downregulated Bcl-2 expression. Therefore, TFPB1 exhibited anti-proliferative and anti-apoptotic effect partly depending on the suppression of Akt signaling pathway. These findings provided us a potential chemotherapeutic strategy for the treatment of human non-small cell lung cancer.

Antioxidant Activity and Oxidative Injury Rehabilitation of Chemically Modified Polysaccharide (TAPA1) from Tremella aurantialba

In our previous work, an acidic homogenous polysaccharide TAPA1 purified from Tremella aurantialba and its acetylated derivative TAPA1-ac, deacetylated derivative TAPA1-deac and sulfated fractions TAPA1-s were obtained by chemical modification and their partially chemical characterization were investigated. In present work, the antioxidant activities of various polysaccharides, including TAPA1, and its three derivative fractions, were determined in terms of scavenging of superoxide anion radicals and hydrogen peroxide (H2O2) by using chemiluminescence measurements, and their activities of oxidative injury rehabilitation were assessed by using PC12 oxidative injury model induced with H2O2. In general, compared with TAPA1, TAPA1-ac showed significant antioxidant activities and higher oxidative injury protection effects, whereas TAPA1-deac and TAPA1-s showed lower effects. These findings seemed to suggest that antioxidant effects and rehabilitation effect of oxidative injury, relate to not only different substituted group, but also the content and the degree of substitution (DS) of acetyl group, indicating that acetylation of TAPA1 was an effective means of enhancing its antioxidant activity and rehabilitation effect.

Fucogalactan from the giant mushroom Macrocybe titans inhibits melanoma cells migration

An aqueous extract containing polysaccharides was obtained from the giant mushroom Macrocybe titans, and it was purified by amylase treatment, freeze-thawing process and dialysis. The purified fraction (ESP) was analyzed by HPSEC and GC–MS which showed a homogenous polysaccharide with Mw 14.2 × 103 g/mol composed by galactose and fucose. NMR and methylation analysis of ESP confirmed the presence of a fucogalactan with a (1 → 6)-linked α-d-Galp main chain partially substituted at O-2 by non reducing end units of α-l-Fucp residues in the side chain. Its biological activity was evaluated against murine melanoma cells B16-F10. The fucogalactan did not alter the viability, proliferative capacity and morphology of cells. However, this polysaccharide was able to reduce the cell migration in vitro at 40% (100 μg/mL) and 33% (250 μg/mL). The results obtained showed that Macrocybe titans fucogalactan is a promising agent capable of altering melanoma cell migration without decrease the cell viability.

The isolation, structural characterization and anti-osteosarcoma activity of a water soluble polysaccharide from Agrimonia pilosa

A homogenous polysaccharide (APP), with a molecular weight of 120 kDa, was isolated from the dried aerial parts of Agrimonia pilosa. Gas chromatography (GC) and GC–MS analysis revealed that APP has a backbone of 1,3-linked Glcp and 1,3, 6-linked Glcp, and branched with 1-linked Glcp terminal along the main chain in a relative ratio of 2:1:1. We investigated the response of human osteosarcoma U-2 OS cells to APP treatment. MTT result showed that APP significantly inhibited cell viability in a concentration dependent manner via induction of apoptotic death in U-2 OS cells, as determined by annexin V/propidium iodide (PI) staining. Western blot analysis also indicated that APP CRA increased in Bax/Bcl-2 ratios by up-regulating Bax expression and triggered the release of cytochrome c from mitochondria into the cytoplasm. Moreover, APP supplement induced the activation of caspase-3, and −9, but not caspase-8 in U-2 OS cells. Likewise, APP administration significantly suppressed tumor growth in BALB/C nude mice bearing U-2 OS xenograft tumors. All these results indicate that APP-induced apoptosis is associated with the activation of a caspase-3-mediated mitochondrial pathway.

Structural Variation and Microrheological Properties of a Homogeneous Polysaccharide from Wheat Germ.

A novel polysaccharide (WGP) was purified from crude wheat germ polysaccharide by Sephacryl S-500HRgel filtration. The molecular weight of WGP was determined as 4.89 × 106 Da and consisted of arabinose, xylose, glucose, and galactose. Methylation analysis and 1D/2D nuclear magnetic resonance was used to analyze the structural characterization of WGP. WGP was mainly a backbone composed of (1 → 4)-linked-β-d-Xylp (19.01%) and (1 → 3, 4)-linked-β-d-Xylp (26.27%) residues, which was branched of (1 → 5)-linked α-l-Araf (28.09%) and (1 → 3,6)-linked β-d-Galp (12.11%) with β-d-Glcp (14.52%) as terminal unit. The calculated values of Turbiscan stability indexes suggested that WGP (0.1-0.5 mg/mL) is a stable system. Microrheology results showed that WGP can form gel behavior when the concentration of WGP ranges from 0.1 to 3 mg/mL. Results of in vitro assays showed that WGP could cause the proliferation of RAW264.7 macrophages, upregulating the release of TNF-α and IL-8 in the lymphocytes.

Preliminary Characterization of a Homogeneous Polysaccharide with Anticomplement Activity from Sijunzi Decoction

Sijunzi decoction (SJZD) is a classical herbal prescription in traditional Chinese medicine (TCM) used for enhancing the function of immune system. In previous studies, a polysaccharide fraction S-3 was screened from SJZD by assessment of immune system regulation, intestinal microbiota, and SCFA in order to explore the immune active ingredients in SJZD. In the present study, S-3 was further purified, and a homogeneous polysaccharide S-3-1 with a molecular mass of 13.5 × 104 Da was obtained after further fractionation by Sephadex G-150 size-exclusion chromatography. The immunological activities of S-3-1 were assayed in vitro for the first time. The determination of the anticomplement activity showed that S-3-1 displayed inhibitory effects on classical pathway of the complement system, with CH50 values of 530 μg/mL. The FT-IR analysis showed that S-3-1 had absorptive peaks characteristic of polysaccharides. The methylation and GC-MS analysis showed that it is comprised of Rha, Ara, Xyl, Man, Gal, and Glc in a relative molar ratio of 0.35 : 0.37 : 1.4 : 0.31 : 3 : 0.8 and that it mainly contained 1,4-linked-Glc and 1,6-linked-Gal glycosidic bonds. The morphology of S-3-1 was observed by atomic force microscope (AFM). These results provided evidences for tracking the material basis of SJZD immune activity.

Progress of the components and biological activities of Morinda officinalis How

The Morinda officinalis How is “one of the top four south authentic traditional Chinese medicines”, widely distributed in South District of China, such as Fujian, Guangdong, Guangxi, Hainan, etc. Its roots are widely used for the treatment of sexual impotence, spermatorrhea, irregular menstruation, and female infertility in clinical. Many kinds of compounds (iridoid glycosides, anthraquinones, saccharides, organic acids, volatile oils and homogeneous polysaccharides) have been isolated from its roots and the relevant biological activities (pain-killing, antioxidant, antibacterial, anticancer, anti-inflammatory, anti-tubercular and cardiovascular action) were also studied. This review briefly describes the botanical description, plant taxonomy, history of medicinal development and the progress of the chemical components and biological activities of M. officinalis to provide a reference for the researchers.

The chemical structure and anti-aging bioactivity of an acid polysaccharide obtained from rose buds.

An acid polysaccharide, named R-PL, was extracted from rose buds by hot water (80 °C) extraction and purified by Sephadex G-200. R-PL, extracted with a total yield of 1.90%, is a highly pure polysaccharide with a total sugar content of 92.51%. A single and symmetrically sharp peak in its high-performance gel-permeation chromatography (HPGPC) spectrum indicates that R-PL is a homogeneous polysaccharide with a molecular weight of 7.727 × 105 Da. Monosaccharide composition analysis shows that it consists of d-arabinose, d-xylose, d-glucose, d-galactose, d-galacturonic acid, and d-glucuronic acid with a molar ratio of 4.6 : 1.4 : 5.22 : 4.81 : 1 : 1.86. Structure analysis, such as FTIR spectroscopy, periodic acid oxidation, Smith degradation, methylation, and NMR analysis, reveals that R-PL has a backbone of →1)-α-d-Glcp-(6→ with side chains of →1)-β-d-Galp-(4,6→, →1)-β-d-Araf-(5→, →1)-α-d-Xylp-(4→, →1)-α-d-GlcpA-(2→ and →1)-α-d-GalpA-(4→. Its morphological characteristics were identified by Congo red experiments and scanning electron microscopy (SEM) analysis. The results show that R-PL does not have a triple-helical conformation in solution; its shape resembles twisted ribbons. Anti-oxidative and anti-aging analyses show that R-PL has significant antioxidant and anti-aging abilities in vivo.

Preliminary characterization and anti-hyperglycemic activity of a pectic polysaccharide from okra (Abelmoschus esculentus (L.) Moench)

Okra is widely used as vegetable, food ingredient, and traditional medicine in China for various applications. The main healthy properties are accepted to originate from the high polysaccharide content of okra. In this study, a homogenous pectic polysaccharide WOP-2 was extracted, isolated, and purified from okra by DEAE-cellulose and Sepharose CL-6B column, and its molecular weight was estimated to be 580 kDa. Monosaccharide composition was analyzed to be Rha (21.4%), GalA (34.9%), Gal (29.6%), GlcA (4.5%), Glc (5.9%), and Ara (3.7%) by HPLC with PMP derivatization. Combined with NMR analysis, WOP-2 was elucidated to be a rhamnogalacturonan I backbone with type-II arabinogalactan side-chains substituted partly at O-4 of Rhap. WOP-2 exhibits significant anti-hyperglycemic activity on STZ-induced diabetic mice by inhibiting the lipid peroxidation chain reaction probably. Overall, these results provide new insight into structure–activity relationships of polysaccharide from okra and provide impetus towards the development of polysaccharide-based therapeutics on diabetes.

Sulfated modification of the polysaccharides from blackcurrant and their antioxidant and α-amylase inhibitory activities

Sulfated modification was conducted to modify a homogenous polysaccharide from blackcurrant (BCP). The sulfated polysaccharides (SBCPs) with different degree of substitution (DS) were synthesized using the aminosulfonic acid (ASA)/4-dimethylaminopyridine method by varying reaction conditions such as the mass ratio of ASA to BCP, temperature, and time. Three sulfated derivatives were chosen for high-performance gel-permeation chromatography, gas chromatography, fourier-transform infrared (FT-IR) spectroscopy, and nuclear magnetic resonance (NMR), and activity studies, designated as SBCP-1, SBCP-2, and SBCP-3 with DS of 1.28, 0.95, and 0.53, respectively. Results showed that the sulfated modification was successful, and SBCPs had an increase in molecular weight compared to BCP. Both SBCPs and BCP were composed of rhamnose, arabinose, xylose, mannose, galactose, and glucose, with different molar ratios. Sulfate substitution was further confirmed by FT-IR and 13C NMR analysis. SBCPs exhibited excellent antioxidant capacities (DPPH, hydroxyl, and superoxide radical scavenging, reducing power, and ferrous metal-chelating capacities) and α-amylase inhibitory activity in vitro, and the activities of SBCPs were significantly improved in positive correlation with the DS value. This study suggested that SBCPs could serve as potential antioxidant agents to be used as alternative supplements or functional foods.

Purification of polysaccharide from artificially cultivated Anoectochilus roxburghii (wall.) Lindl. by high-speed counter current chromatography and its antitumor activity.

To establish a systematic method for the extraction, purification, characterization and antitumor activity study of polysaccharide from artificially cultivated Anoectochilus roxburghii (wall.) Lindl. (AC-ARPS). High-speed counter current chromatography with two-phase aqueous systems was successfully applied to purify AC-ARPS after one-step separation. The purity of the AC-ARPS obtained by phenol/sulfuric acid method was 95.01%. The chemical structures of AC-ARPS were identified by a series of analytical methods including high-performance liquid chromatography and liquid chromatography with mass spectrometry. High-performance liquid chromatography and liquid chromatography with mass spectrometry indicated that AC-ARPS was mainly composed of mannose, ribose, glucose, galactose and arabinose with a molar ratio of 1.00:8.47:47.30:1.17:1.19. AC-ARPS is a homogeneous polysaccharide with a molecular weight of 25 681Da. The antitumor effect of AC-ARPS was evaluated on lung cancer A549, osteosarcoma 143B, rat adrenal pheochromocytoma PC 12, breast cancer MCF-7, acute leukemia HL 60, chronic leukemia K562, colon cancer SW620, esophageal cancer OE 19, liver cancer HepG2, and neuroglioma U251 cells in vitro. AC-ARPS showed the best inhibitory effect on OE 19 cells, and the IC50 value was 5.67 ± 0.831 μmol/L. Fluorescence analysis and flow cytometry results showed that AC-ARPS induced apoptosis and G2/M phase arrest in OE 19 cells.

Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds

Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC–MS), as well as nuclear magnetic resonance (NMR) spectra. The weight-average molecular weight for CP-40-M was determined to be 0.29×105Da, and the molar ratio of xylose to glucuronic acid was 4.62. The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. It was the first time to obtain this type of xylan from the water extract of Cassia obtusifolia seeds. The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine.

Structural elucidation and antiaging activity of polysaccharide from Paris polyphylla leaves

To optimize the use of Paris polyphylla resources, a homogenous polysaccharide (PPLP) was obtained from P. polyphylla leaves. Its average molecule weight was 2.95×104Da, and the analysis of monosaccharide composition shown that PPLP consisted of l-arabinose and d-galactose with a molar ratio of 4.2:5.8. Methylation and nuclear magnetic resonance (NMR) spectroscopy data revealed that the backbone of PPLP was comprised of (1→6)-β-d-galactan, and the branched chains mainly consisted of arabinosyl residues which was linked to backbone via (1→3)-linkages. In addition, the antiaging effect of PPLP was investigated in a d-galactose induced mouse aging model. Compared with model group, the formations of malondialdehyde (MDA) were significantly prevented, and the levels of antioxidant enzymes and total antioxidant capacity (TAOC) were significantly improved in serum and liver in PPLP dose groups. These results demonstrated that PPLP possessed potent antiaging capacity.

Anti-complementary activity of two homogeneous polysaccharides from Eclipta prostrata

Complement-mediated inflammation and tissue damage is an important drive to pathology in autoimmune diseases, targeting inhibit the complement activation is promising treatment strategy for these diseases. We performed anticomplement activity-guided fractionation of the water extract of Eclipta prostrata by ion-exchange and size-exclusion chromatography, yielding two bioactive polysaccharides EAP20-1 and EAP20-2. The molecular weights of EAP20-1 and EAP20-2 were respectively calculated to be 5.2 kDa and 6.3 kDa by HPGPC, both polysaccharides was composed by d-Gal, l-Glc, and Ara at different ratios of 7.3:2.7:1 and 7.6:3.1:1. In addition, the main linkage types of EAP20-1 and EAP20-2 were β-1,4-Gal, β-1,6-Gal and α-1,4,6-Glc according to methylation analyses. EAP20-1 and EAP20-2 exhibited significant inhibitory effect on the complement activation through both classical and alternative pathways and with no influence on the coagulation system. Preliminary mechanism study indicated that both EAP20-1 and EAP20-2 inhibited the activation of the complement system by interacting with C1q, C1r, C1s, C2, C4, C5, C7, and C9 components.

Purification, characterization and immunomodulatory activity of water extractable polysaccharides from the swollen culms of Zizania latifolia

The swollen culms of Zizania latifolia is a popular vegetable and traditional herbal medicine mainly consumed in some Asian countries. In our previous study, water extractable polysaccharide (ZLPs-W) and alkali extractable polysaccharide (ZLPs-A) was sequentially prepared from the swollen culms of Zizania latifolia. In vitro cell assay revealed that ZLPs-W without cytotoxicity had higher immunomodulatory activity than ZLPs-A. Therefore, in this study, ZLPs-W was further separated to three purified polysaccharides of ZLPs-W1, ZLPs-W2, and ZLPs-W3 by anion exchange chromatography and size exclusive chromatography in sequence. They were found to be homogeneous polysaccharides in β-type glycosidic linkage. ZLPs-W1 and ZLPs-W2 without triple helix conformation mainly contained Glc in molar percentage of 96.47% and 85.1%, respectively. ZLPs-W3 with triple helix conformation mainly consisted of Man, Rha, GalA, Glc, Gal and Ara in the molar ratio of 10.23:14.14:37.44:17.14:16.54:4.51. In addition, all the three purified polysaccharides could effectively enhanced the proliferation, phagocytosis, and nitric oxide production of murine macrophage RAW 264.7, suggesting that they possessed a potent immunostimulatory activity and could be developed as immunomodulator in medicine or functional food areas.

Digestive behavior of Dendrobium huoshanense polysaccharides in the gastrointestinal tracts of mice

With fluorescent end-labeling method, a homogenous Dendrobium huoshanense polysaccharide (GXG) that was orally administrated to mice was found to be degraded into a stable fragment (dGXG) in the stomach, where dGXG was propelled into the small intestine and absorbed into the systemic circulation. The residual GXG in the stomach could pass through the gastrointestinal tract and was excreted into faeces with a very slow fermentation in the large intestine. The simulated digestion of GXG indicated that the acidic pH condition in the gastric fluid was responsible for GXG degradation, which was supported by the oral digestion of dGXG. Chemical analysis not only showed a change in the molar ratio of monosaccharide compositions and glycosidic linkage types, but also found the loss of the sugar residue (1→4)-linked Galp after the acid treatment of GXG, suggesting that the cleavage of glycosidic linkage, especially (1→4)-linked Galp linkage, resulted in GXG degradation in the gastrointestinal tract.

Characterization and immunomodulatory activity of polysaccharides from the stems and leaves of Abelmoschus manihot and a sulfated derivative

BackgroundAbelmoschus manihot (Linn.) Medicus is a traditional herbal medicine whose flowers, stems and leaves exhibit widely pharmacological activities. However, only the flowers have long been used as medicine while the stems and leaves were mainly discarded and burned, which undoubtedly caused enormous waste of these resources and serious environment pollution. Many researches have indicated that bioactivities of polysaccharides were significantly improved after sulfation. The aim of this study was to investigate the characterization and immunomodulatory activity of polysaccharides from stems and leaves of A. manihot and a sulfated derivative. ResultsA mixed neutral polysaccharide (SLAMP-a) and two acidic polysaccharides (SLAMP-c and SLAMP-d) were obtained from stems and leaves of A. manihot by DEAE-cellulose chromatography. SLAMP-a was a water-insoluble mixture while its sulfated derivative (S-SLAMP-a3), prepared with aminosulfonic acid, was a homogeneous polysaccharide with excellent solubility. The average molecular weights of S-SLAMP-a3, SLAMP-c and SLAMP-d were 1044.2kDa, 477.8kDa and 264.2kDa respectively. SLAMP-a and its sulfate mainly contained glucose, and SLAMP-c and SLAMP-d were both composed of mannose, rhamnose, glucuronic acid, glucose, galactose, and arabinose. In vitro study indicated that S-SLAMP-a3, SLAMP-c and SLAMP-d exhibited significant immunomodulatory activity, while SLAMP-a showed little effects. ConclusionSLAMP-c and SLAMP-d from A. manihot stems and leaves could be explored as immunomodulatory agents, which would provide a way to utilize these enormously discarded resources and avoid massive waste. Additionally, the neutral polysaccharide, SLAMP-a, could also be developed after sulfation, suggesting that these disused resources would be further used effectively.

Purification, characterization and biological effect of reversing the kidney-yang deficiency of polysaccharides from semen cuscutae

Semen cuscutae is a well-known Chinese medicine which has been used to nourish kidney. It is the first study to demonstrate that the polysaccharides from semen cuscutae showed significant activity of nourishing kidney-yang by increasing the levels of testosterone and estradiol, decreasing the level of blood urea nitrogen, improving immune function, possessing antioxidant effect. Three homogeneous polysaccharides were obtained by DEAE-cellulose and Sephacryl S-400 which were named as C-7WR1, C-7WR2 and C-7WR3 with average molecular weight of 7.59×104, 3.23×104 and 2.25×104 respectively. C-7WR1 was composed of fructose: mannose=0.02:1. C-7WR2 was composed of fructose: mannose: xylose: arabinose=0.01:1:0.14:0.33. C-7WR3 was composed of fructose: mannose: xylose: arabinose=0.01:1:0.10:0.47. They mainly contained mannose. Their fourier transform infrared features were similar. They all had no nucleic acid and protein.

Structural analysis and immunomodulatory effect of polysaccharide from Atractylodis macrocephalae Koidz. on bovine lymphocytes

RAMP60, RAMP70, RAMP80 and RAMPtp extracted from the rhizome of Atractylodis macrocephalae Koidz. were chemically analyzed and tested for their lymphocyte stimulation. Among them, only RAMPtp was found to induce proliferation of lymphocytes from bovine supramammary lymph node (SMLN). It was found that RAMPtp was homogeneous polysaccharide with Mn of 1.867×103 by GPC analysis, composed of glucose, mannose, rhamnose, arabinose and galactose, with mass percentages of 60.67%, 14.99%, 10.61%, 8.83% and 4.90%, respectively by HPLC analysis, connected by 1, 3-linked β-D Galp and 1, 6-linked β-D Galp residues by NMR analysis and was amorphous in structure by SEM analysis. When SMLN lymphocytes were cultured with RAMPtp in vitro, increased [Ca2+]i, more cells in S and G2/M phases and upregulated IFN-γ and IL-17A while downregulated IL-4 mRNA expressions as well as the binding between SMLN lymphocytes and RAMPtp were observed. These findings might explain the lymphocyte-stimulating property of RAMPtp.

Structural characterisation of a water-soluble polysaccharide from tissue-cultured Dendrobium huoshanense C.Z. Tang et S.J. Cheng

A water-soluble polysaccharide TC-DHPA4 with a molecular weight of 8.0 × 105 Da was isolated from tissue-cultured Dendrobium huoshanense by anion exchange and gel permeation chromatography. Monosaccharide analysis revealed that the homogeneous polysaccharide was made up of rhamnose, arabinose, mannose, glucose, galactose and glucuronic acid with a molar ratio of 1.28:1:1.67:4.71:10.43:1.42. The sugar residue sequence analysis based on the GC-MS files and NMR spectra indicated that the backbone of TC-DHPA4 consisted of the repeated units:→6)-β-Galp-(1→6)-β-Galp-(1→4)-β-GlcpA-(1→6)-β-Glcp-(1→6)-β-Glcp-(→. The sugar residue sequences β-Glcp-(1→)-α-Rhap-(1→3)-β-Galp-(1→, β-Glcp-(1→4)-α-Rhap-(1→3)-β-Galp-(1→, β-Galp-(1→6)-β-Manp-(1→3)-β-Galp-(1→, and α-l-Araf-(1→2)-β-Manp-(1→3)-β-Galp-(1→ were identified as the branches attached to the C-3 position of (1→6)-linked galactose in the backbone.