Optimization of the fermentation media, mathematical modeling, and enhancement of paclitaxel production by Alternaria alternata after elicitation with pectin.

Alternaria alternata fungus is a potent paclitaxel producer isolated from Corylus avellana. The major challenge is the lack of optimized media for endophytic fungi productivity. In the effort to maximize the production of taxoids by A. alternata, several fermentation conditions, including pH (pH 4.0-7.0), different types and concentrations of carbon (fructose, glucose, sucrose, mannitol, sorbitol, and malt extract), and nitrogen (urea, ammonium nitrate, potassium nitrate, ammonium phosphate, and ammonium sulfate) were applied step by step. Based on the results, A. alternata in a medium containing sucrose 5% (w/v) and ammonium phosphate 2.5 mM at pH 6.0 showed a rapid and sustainable growth rate, the highest paclitaxel yield (94.8 µg gFW-1 vs 2.8 µg gFW-1 in controls), and the maximum content of amino acids. Additionally, the effect of pectin was evaluated on fungus, and mycelia harvested. Pectin significantly enhanced the growth and taxoid yield on day 21 (respectively 171% and 116% of their corresponding on day 7). The results were checked out by mathematical modeling as well. Accordingly, these findings suggest a low-cost, eco-friendly, and easy-to-produce approach with excellent biotechnological potential for the industrial manufacture of taxoids.

Grafting of sinapic acid onto glucosamine nanoparticle as a potential therapeutic drug with enhanced anti-inflammatory activities in osteoarthritis treatment.

Glucosamine (Glu) is a cartilage and joint fluid matrix precursor that modulates osteoarthritic joint changes. To improve the enzymatic stability, glucosamine was developed into nanoglucosamine by the ionic gelation method through sodium tripolyphosphate (TPP) as cross-linking agent. The optimized mass ratio of Glu:TPP was (3:1) with the particle size 163 ± 25 nm and surface charge -5 mV. Then Sinapic acid (SA) as a natural phenolic acid with strong antioxidant and antimicrobial activities has been grafted onto glucosamine nanoparticles (GluNPs) with grafting efficiency (73 ± 6 %). The covalent insertion of SA was confirmed by UV-Vis, FTIR, 1HNMR, XRD, and FESEM analyses and the other physicochemical properties were also characterized. SA-g-GluNPs showed spherical shape with a mean diameter of 255 ± 20 nm and zeta potential +16 mV. The in vitro release profile of SA-g-GluNPs exhibited the sustained and pH-dependent drug release property. SA-g-GluNPs had a more pronounced effect on reducing the elevated levels of LPS-induced oxidative stress and pro-inflammatory cytokines than free SA in the human chondrocyte C28/I2 cell line. Furthermore, the antibacterial properties against E. coli and S. aureus were also improved by SA-g-GluNPs. This study demonstrated the potential of phenolic acid grafted GluNPs in therapeutic drug applications for chondroprotection and food industries.

Structure-activity relationship of fucoidans and alginates obtained from Cystoseira indica in a biorefinery concept.

A sequential extraction process was employed to isolate fucoidan and alginate from brown seaweed Cystoseira indica. Extraction process was designed to evaluate the effects of acid concentrations (0.025, 0.05, 0.1 and 0.2 M HCl) and temperatures (room temperature, 60 °C and 80 °C) on sensory, structural and immunostimulatory properties of fucoidans and following results on Na+-alginates. The amounts of isolated fucoidans (0.193-0.658 g/5 g powder) and Na+-alginates (2.877-3.383 g/ 5 g powder) greatly varied among different extractions. Fucoidans were composed of neutral sugars, mainly fucose (15.74-47.64 %) and galactose (18.66-26.88 %) units, with varying amounts of sulfates (8.76-12.40 %) and uronic acids (0.46-8.90 %). The weight average molecular weights (Mw) of fucoidans (234.6-1990.0 × 103 g/mol) and Na+-alginates (358.4-2318.3 × 103 g/mol) were closely controlled by extraction condition. Both fucoidan and Na+-alginate molecules noticeably induced RAW264.7 murine macrophage cells to exert proinflammatory response, producing considerable levels of NO, IL-1ß, TNF-α and IL-6 through NF-κB and MAPKs signaling pathways. Altogether, extraction process of fucoidan not only exerted determining effect on its structure and cell activation capacity, but also influenced the quality of Na+-alginate obtained in the next step.

Ultrasonic-assisted enzymatic extraction of sulfated polysaccharide from Skipjack tuna by-products.

The effect of ultrasound pretreatment on extraction efficiency of sulfate polysaccharides (SPs) using alcalase from different by-products of Skipjack tuna including head, bone and skin was evaluated. Structural, functional, antioxidant and antibacterial properties of the recovered SPs using the ultrasound-enzyme and enzymatic method were also investigated. Ultrasound pretreatment significantly increased the extraction yield of SPs from all the three by-products compared with the conventional enzymatic method. All extracted SPs showed high antioxidant potential in terms of ABTS, DPPH and ferrous chelating activities where the ultrasound treatment enhanced antioxidant activities of the SPs. The SPs exerted strong inhibiting activity against various Gram-positive and Gram-negative bacteria. The ultrasound treatment remarkably increased antibacterial activity of the SPs against L. monocytogenes but its effect on other bacteria was dependent on the source of the SPs. Altogether, the results suggest that ultrasound pretreatment during enzymatic extraction of SPs from tuna by-products can be a promising approach to improve extraction yield but also bioactivity of the extracted polysaccharides.

Fish Protein Hydrolysate from Sulfated Polysaccharides Extraction Residue of Tuna Processing By-Products with Bioactive and Functional Properties.

The ethanol-induced precipitation after enzymatic hydrolysis commonly used for sulfated polysaccharide extraction from marine resources wastes a large amount of proteins. Here, possible extraction of fish protein hydrolysates (FPH) from the ethanol residue of sulfated polysaccharide precipitation from head, bone, and skin of skipjack tuna is investigated. Antioxidant, antibacterial, angiotensin I-converting enzyme (ACE) inhibitory activities and functional properties of the recovered FPHs are also evaluated. A degree of hydrolysis of 40.93, 38.13, and 37.23 is achieved for FPH from head, bone, and skin, respectively. FPH from the head presents the highest antioxidant and ACE inhibitory activity as well as foam/emulsion capacity among all the FPHs. The FPHs are all able to inhibit three Gram-positive bacteria and three Gram-negative bacteria to varying degrees and have a water solubility >65%. Altogether, the results demonstrate great potential for recovery of bioactive/functional peptides from the residue of sulfated polysaccharide extraction process enabling efficient biorefining of aquatic resources.

Immunostimulatory effects of a polysaccharide from Pimpinella anisum seeds on RAW264.7 and NK-92 cells.

Polysaccharides from Pimpinella anisum were isolated using water at elevated temperature and DEAE Sepharose FF chromatography to examine their chemical structure and activation capacity on immune cells. P. anisum fractions (PAF1, PAF2 and PAF3) were mainly composed of neutral sugars (84.0-98.2%) and uronic acids (2.1-11.8%) with weight average molecular weight (Mw) ranging from 186.6 to 5474.5 × 103 g/mol. Polysaccharides induced a significant inflammatory response in RAW264.7 murine macrophage cells releasing nitric oxide and expressing TNF-α, IL-1ß, IL-6 and IL-10 cytokines. The induction of NK-92 natural killer cells resulted in TNF-α and IFN-γ production and activation of GrB/perforin-, NKG2D- and FasL-mediated cytotoxicity. Polysaccharides triggered the phosphorylation of NF-κB, ERK, JNK and p38 proteins in RAW264.7 and NK-92 cells indicating the involvement of NF-κB and MAPKs signaling pathways. The most active polysaccharide was a galactoarabinan with complex structure.

Purification, characterization and immunostimulatory effects of polysaccharides from Anemarrhena asphodeloides rhizomes.

The crude polysaccharide was extracted from A. asphodeloides rhizomes and further purified to produce two fractions F1 (50.0%) and F2 (19.6%). The chemical constitutions of the polysaccharides were neutral sugars (51.4%-89.7%), uronic acids (1.0%-30.2%) and sulfate esters (3.4%-8.1%), with various ratios of monosaccharides including rhamnose (1.4%-6.1%), arabinose (7.1%-21.2%), xylose (0.2%-4.8%), mannose (39.9%-79.0%), glucose (6.0%-11.1%) and galactose (2.6%-22.0%). The molecular properties of the polysaccharides were investigated by the HPSEC-UV-MALLS-RI system, revealing the Mw 130.0 × 103-576.5 × 103 g/moL, Rg 87.6-382.6 nm and SVg 0.3-54.3 cm3/g. The polysaccharides stimulated RAW264.7 cells to produce considerable amounts of NO and up-regulate the expression of TNF-α, IL-1 and COX-2 genes. Polysaccharides exhibited the growth inhibitory effects on cancer cells lines of AGS, MKN-28 and MKN-45, in which F2 fraction exhibited prominent bioactivities. The AGS cells treated with F2 experienced condensed cytoplasm, shrinkage of nucleus and chromatin marginalization with the highest number of cells at early-stage apoptosis reaching 54.6%. The inhibitory effect of F2 polysaccharide on AGS cells was through MAPKs and STAT3 signaling pathways. The backbone of the F2 was mainly linked by (1 â†’ 4)-linked mannopyranosyl and (1 â†’ 3)-linked galactopyranosyl. Taken together, the polysaccharide from A. asphodeloides rhizomes could be utilized as medicinal, pharmacological and functional food ingredients.

Bioengineered gold nanoparticles from marine seaweed Acanthophora spicifera for pharmaceutical uses: antioxidant, antibacterial, and anticancer activities.

In this study, we report a green synthesis of pharmaceutically active gold nanoparticles from marine red alga Acanthophora spicifera by the reduction of chloroauric acid. The formation of A. spicifera-mediated gold nanoparticles (As-AuNPs) was characterized by several analytical techniques. The crystalline and face-centered cubic (fcc) structure were confirmed by X-ray diffraction (XRD) analysis. Electron microscopy results confirmed that As-AuNPs were spherical and the average size of particles was < 20 nm. As-AuNPs hold a significant level of antioxidant activities than A. spicifera extract. As-AuNPs exhibited the highest antibacterial activity against Vibrio harveyi than Staphylococcus aureus at 100 µg/ml. Furthermore, As-AuNPs exhibited the utmost cytotoxicity against human colon adenocarcinoma (HT-29) cells and registered the half-maximal inhibitory concentration (IC50) at 21.86 µg/ml. These findings authenticated that the synthesized As-AuNPs possess a broad spectrum of biological activities, and it can be effectively applied in the field of aquaculture and biomedical application.

Extraction, structural elucidation and immunostimulating properties of water-soluble polysaccharides from wheat bran.

A water-soluble polysaccharide was extracted from wheat bran (WBP) and investigate their structural characteristics and immunostimulatory activities. The chemical composition of WBP and purified fraction (WBP-F) mainly consists of neutral sugars (91.2 ± 1.2 and 98.7 ± 1.2%), proteins (8.6 ± 0.3 and 0.2 ± 0.1%) and uronic acids (0.7 ± 0.1 and 0.6 ± 0.1%). The molecular weight (Mw ) of WBP and WBP-F was calculated as 911.7 and 510.2 × 103  g/mol, respectively. The WBP-F stimulates the RAW264.7 cells through the production of nitric oxide and various cytokines. The treatment of WBP-F facilitated the phosphorylation of P38, JNK, ERK, and NF-ƘB in RAW264.7 cells suggesting that they might stimulate RAW264.7 cells through the activation of NF-ƙB and MAPKs pathways. Furthermore, the structural details of WBP-F were studied by GC-MS and NMR spectrum, which confirms that the main backbone consists of 4-α-D-linked glucopyranosyl residues with branching points at C-6. PRACTICAL APPLICATIONS: Wheat bran is a potential source of health-promoting compounds. It has been reported that polysaccharides of wheat bran containing numerous beneficial activities. In this study, the wheat bran polysaccharide was extracted, fractionated and investigated their immunostimulatory activities. The results found in this study revealed that the purified polysaccharide from wheat bran potentially enhanced the RAW264.7 cells activation. Hence, these polysaccharides could be utilized as a potent immunity-enhancing agent in food and pharmaceutical industries.

Inducing inflammatory response in RAW264.7 and NK-92 cells by an arabinogalactan isolated from Ferula gummosa via NF-κB and MAPK signaling pathways.

The polysaccharide isolated from F. gummosa (FGP) was found homogenous with a weight average molecular weight (Mw) of 50.0 × 103 g/mol and radius of gyration (Rg) of 105.3 nm. The FGP was an arabinogalactan with a backbone formed of →6)-ß-Galp-1→ residues having random branching points at C-3 extended with either ß-Galp-(1→3)-ß-Galp-(1→ or α-Araf-(1→ side chain residues. FGP exhibited proliferative effect on RAW264.7 cells and induced macrophages to exert proinflammatory response releasing NO and up-regulating the transcription of cytokines including TNF-α, IL-1ß, IL-6 and IL-12. The FGP induced NK-92 cells to up-regulate the expressions of TNF-α, IFN-γ, granzyme-B, perforin, NKG2D and FasL. The presence of p-NF- κB, p-ERK, p-JNK and p-p38 in RAW264.7 and NK-92 cells indicated their activation through NF-κB and MAPKs signaling pathways. These findings suggested that polysaccharides from F. gummosa are potent in boosting immune system and thus may be considered for further studies of biomedical applications.

Sulfated galactan from Halymenia dilatata enhance the antioxidant properties and prevents Aeromonas hydrophila infection in tilapia fish: In vitro and in vivo study.

The structural characterization and pharmaceutical perspective of sulfated galactan from Halymenia dilatata (Hd-SG) were reported in this study. The Hd-SG consists of carbohydrate (58.5 ± 0.9%), sulfate (28.7 ± 0.9%) and protein (2.7 ± 0.2%). The existence of carbon (28.14%), hydrogen (5.50%), nitrogen (0.51%) and sulfur (8.26%) was confirmed in CHNS analysis. The Hd-SG was mainly comprising of galactose and mannose connected by (1 â†’ 4)-glycosidic linkages, and it shows the molecular weight of 900.9 × 103 g/mol in high-performance size exclusion chromatography (HPSEC). The Hd-SG exhibited the dose depended on antioxidant activities. The in vitro and in vivo studies proved the antibacterial efficacy of Hd-SG against Aeromonas hydrophila. The pre-treated Oreochromis fish with Hd-SG (2.0 g/0.1 kg of feed) showed the highest survival, antioxidant, and improved histological changes than the fish infected with A. hydrophila alone. These results concluded that the isolated Hd-SG has extensive therapeutic properties, and it can be used as preventive medicine.

The activation of NF-κB and MAPKs signaling pathways of RAW264.7 murine macrophages and natural killer cells by fucoidan from Nizamuddinia zanardinii.

Polysaccharides from Nizamuddinia zanardinii were extracted using water at elevated temperature and fractionated by a DEAE Sepharose FF column yielding four fractions (F1-F4). Crude and fractions were composed of neutral sugars (50.8-57.4%), proteins (10.8-18.1%), sulfates (7.5-17.3%) and uronic acids (3.5-7.7%). Various levels of galactose (13.4-44.4%), fucose (34.1-40.1%), mannose (14.1-33.2%) and xylose (7.4-15.2%) formed the building blocks of the polysaccharide structures. The weight average molecular weights (Mw) of polysaccharides varied between 40.3 and 1254.4 × 103 g/mol. F3 polysaccharide was the most active fraction stimulating RAW264.7 murine macrophage cells to secrete NO, TNF-α, IL-1ß and IL-6, and activating NK cells to release TNF-α, INF-γ, granzyme-B, perforin, NKG2D and FasL through NF-κB and MAPKs signaling pathways. Highly-branched F3 polysaccharide mainly consisted of (1 â†’ 2)-Fucp, (1 â†’ 2,3)-Manp, (1 â†’ 3)-Galp, (1 â†’ 2)-Manp, (1 â†’ 3)-Manp, (1 â†’ 2,3,4)-Manp and (1 â†’ 2,3,6)-Manp residues with great amount of (→1)-Fucp and (→1)-Xylp. Sulfates substituted at C-2 of fucose and galactose residues. Overall, fucoidan from N. zanardinii showed immense potency in boosting immune system through macrophages and NK cells activations and therefore suitable for further exploration in immune-mediated biomedical applications.

Isolation, structural elucidation and immuno-stimulatory properties of polysaccharides from Cuminum cyminum.

The aim of the present study was to evaluate the effect of water-soluble polysaccharides from Cuminum cyminum to induce inflammatory response in immune cells and understand their underlying mechanisms. Weight average molecular weight (Mw) of polysaccharides varied between 191.4-512.2 × 103 g/mol. Polysaccharides induced RAW264.7 cells to release nitric oxide and express TNF-α, IL-1ß, IL-6 and IL-12 inflammatory cytokines. Polysaccharides activated NK-92 cells to produce TNF-α, IFN-γ, perforin, granzyme B, NKG2D and FasL. Activations of RAW264.7 and NK-92 cells were through NF-κB and MAPKs signal pathways indicated by the presence of phosphorylated NF-κB, ERK, JNK and p38 proteins. The polysaccharide structure was mainly constituted of →4)-Galp-(1→, →3)-Galp-(1→, →2)-Arap-(1→ and →2)-Arap-(1→ glycosidic linkages. Overall results suggested that polysaccharides from C. cyminum possessing lower MW and greater expanded conformation more effectively stimulate RAW264.7 and NK-92 cells and thus could be considered for further studies on their biomedical applications.

Structural characteristics, molecular properties and immunostimulatory effects of sulfated polysaccharide from freshwater Myriophyllum spicatum L.

Water-soluble polysaccharides were isolated from a freshwater plant, Myriophyllum spicatum L., at elevated temperature and purified using DEAE Sepharose FF column to elucidate their structural properties and immunopotentiating effects. Polysaccharides were composed of neutral sugars (73.9-78.2%), sulfate esters (12.8-16.8%) and uronic acids (3.1-4.9%). Weight average molecular weight (Mw) of polysaccharides varied between 217.4 and 529.0 × 103 g/mol. Polysaccharide structures were predominantly consisted of glucose (31.9-59.1%) and galactose (16.6-34.5%) with lower amounts of mannose, xylose, arabinose and rhamnose. As the most active fraction, MSF1 induced RAW264.7 murine macrophage cells to release nitric oxide and initiate the expression of TNF-α, IL-1ß, IL-6 and IL-12 cytokines. Besides, MSF1 activated NK-92 cells to produce TNF-α and INF-γ cytokines and initiate cell-mediated cytotoxicity via perforin/granzyme B, NKG2D receptor and FasL pathways. Stimulation of RAW264.7 and NK-92 cells with MSF1 resulted in high expressions of phosphorylated NF-κB, ERK, JNK and p38 proteins revealing the involvement of NF-κB and MAPKs signaling pathways. MSF1 possessed a highly-branched structure consisting a combination of (1 → 4)-Glcp, (1 → 6)-Glcp, (1 → 4,6)-Glcp, (1 → 3,4)-Galp and (1 → 2,3)-Galp residues. Altogether, the present findings suggested the potential application of sulfated-polysaccharides from M. spicatum L. for immunostimulation and thus the necessity for further biomedical research.

Isolation and structural characterization of sulfated polysaccharide from Spirulina platensis and its bioactive potential: In vitro antioxidant, antibacterial activity and Zebrafish growth and reproductive performance.

In this study, the sulfated polysaccharide (SPs) was isolated from Spirulina platensis. The isolated SPs contains carbohydrate, sulfate, protein and uronic acid at 38.7 ±â€¯0.30%, 21.3 ±â€¯0.87%, 7.1 ±â€¯0.15% and 7.9 ±â€¯0.4% respectively. The elemental analysis confirmed the presence of carbon (18.01 ±â€¯0.10%), hydrogen (1.83 ±â€¯0.02%) and nitrogen (3.43 ±â€¯0.01%). The monosaccharide composition and molecular weight of SPs were analyzed by high-performance liquid chromatography and size exclusion chromatography respectively. The monosaccharide composition analysis showed the existence of glucose, rhamnose, xylose, fucose, mannose, galactose and the molecular weight of SPs was 1016 kDa. Further, the characterization of SPs was done by UV-visible spectroscopy, X-ray diffraction, FT-IR, 1H NMR and 13C NMR analysis. The obtained SPs exhibited potent antioxidant activity in DPPH (76.45 ±â€¯0.49%), reducing power (absorbance: 1.3 ±â€¯0.02), hydrogen peroxide scavenging (66.3 ±â€¯1.16%), hydroxyl scavenging (68.6 ±â€¯3.2%), nitric oxide (81.36 ±â€¯1.85%) and total antioxidant (absorbance:1.66 ±â€¯0.02) activities at 5 mg/ml. In addition, SPs revealed the highest antibacterial efficacy against the pathogenic bacteria Vibrio vulnificus in disc diffusion, agar bioassay and protein leakage assays at 100 µg/ml. Furthermore, the supplementation of 2% SPs through a feed to the Danio rerio fish enhances the growth and reproductive performances. This finding confirmed that the isolated SPs from S. platensis possess pharmaceutical as well as nutritional properties.

Addition of seaweed powder and sulphated polysaccharide on shelf_life extension of functional fish surimi restructured product.

This study aimed to investigate the performance of equal amounts of edible green seaweed, Ulva intestinalis powder (2.77 g kg-1), and its sulphated polysaccharide ([USP], 0.5 g kg-1, based on the extraction yield from U. intestinalis powder) on the proximate compositions, lipid oxidation, pH, colour, textural properties, cooking yield and sensory attributes of fish-surimi restructured products during storage at - 18 °C as compared with the control. Results showed incorporation of two functional components resulted in lower TBARS values compared with the control over 6 months (P ≤ 0.05). The USP incorporated fingers showed the least moisture loss over 6 months (P < 0.05). Textural properties for two functional fingers remained relatively stable from month 0 to month 6, while the hardness increased significantly (P < 0.05) in the control fingers (67 to 80 N). Additionally, the sensory attributes of all formulated fingers were judged acceptable; however, the USP containing fingers were preferred by the sensory panelists, due to their juicy texture as a result of less cooking loss comparing with others. In conclusion, this study suggests the potential use of such natural marine ingredients to maintain the quality and to extend the shelf life of surimi-based products with beneficial health effects.

The activation of RAW264.7 murine macrophage and natural killer cells by glucomannogalactan polysaccharides from Tornabea scutellifera.

A water-soluble polysaccharide was isolated from Tornabea scutellifera and fractionated using a DAEA Sepharose FF column to evaluate its capacity to stimulate natural killer (NK) cells and macrophages. Neutral sugars (71.8-93.5%) constituted the major part of crude polysaccharides and fractions (TSF1 and TSF2) with relatively lower levels of proteins (0.4-20.3%) and uronic acids (0.8-4.9%). The weight average molecular weights (Mw) of 152.7-537.3 × 103 g/mol were measured for isolated polysaccharides. The polysaccharides were composed of glucose (14.4-44.0%), galactose (23.2-43.2%), mannose (28.5-34.2%) and rhamnose (2.6-13.9%) units connected through (1→2)-Galp, (1→2,6)-Galp, (1→4)-Glcp, (1→6)-Glcp, (1→3)-Rhap, (1→2)-Rhap and (1→4)-Manp residues. TSF2 polysaccharide effectively induced RAW264.7 murine macrophages to release nitric oxide, TNF-α, IL-1ß and IL-6, and activated NK cells to produce TNF-α, INF-γ, granzyme-B, perforin, NKG2D and FasL through NF-κB and MAPKs signaling pathways. Overall results suggested that polysaccharides from T. scutellifera could be potent immunostimulatory compounds inducing both macrophages and NK cells.

Bioactivities of Nizamuddinia zanardinii sulfated polysaccharides extracted by enzyme, ultrasound and enzyme-ultrasound methods.

Sulfated polysaccharide (fucoidan) was isolated from Nizamuddinia zanardinii by enzyme (alcalase), ultrasonic and enzyme-ultrasonic methods. The extracted fucoidans were assessed for their chemical compositions, molecular characteristics, anticancer and immunomodulatory activities. Enzyme-ultrasonic isolated fucoidan showed the maximum extraction yield (7.87%) while that obtained by ultrasonic had the minimum value (3.6%). fucoidans were composed of different levels of carbohydrates (52.78-58.65%), proteins (6.98-8.91%), sulfates (21.78-29.6%) and uronic acids (0.42-1.08%). The weight mean average molecular weight of fucoidans varied between 443.7 and 1020.85 kDa. The polysaccharide chains were consisted of fucose, galactose, glucose, mannose and xylose. All the recovered fucoidans showed strong growth inhibition against HeLa and Hep-G2 cancer cells. The isolated fucoidans were non-toxic and considerably stimulated the macrophage cells to release nitric oxide. Enzyme extraction produced fucoidan with the most macrophage stimulation capacity (> 42 µmol). These results suggested that enzyme treatment preserved more sulfate groups in fucoidan structure influencing its anticancer and immunostimulatory activities.

Subcritical water extraction as an efficient technique to isolate biologically-active fucoidans from Nizamuddinia zanardinii.

A sulfated polysaccharide (fucoidan) has been isolated from Nizamuddinia zanardinii using subcritical water extraction method (SCWE), and extraction conditions were optimised using the response surface methodology. The optimum extraction conditions were found to be: extraction time of 29 min, extraction temperature of 150 °C, and raw material-to-water ratio of 21 g/mL. The fucoidan yield under these optimum conditions was 25.98%, which was considerably higher than that of conventional solvent extraction (5.2%). Extraction time and temperature were the extraction variables that most significantly affected fucoidan yield. Chemical and monosaccharide composition, molecular weight, and the antioxidant, anticancer and immunomodulatory activities of the extract have also been investigated. The monosaccharide composition of fucoidan included fucose (34.13%), mannose (30.70%), galactose (23.19%), xylose (9.35%) and glucose (2.65%). The average molecular weight of the extracted fucoidan was 694 kDa. Antioxidant results revealed that SCWE-extracted fucoidan had appreciable ABTS radical scavenging (70.35%) and reducing power (0.182 Abs). The anticancer activity of fucoidan ranged from 24.60 to 49.46% for HeLa cells and from 23.95 to 46.78% for HepG2 cells. The NO production of RAW264.7 cells was observed to be dose-dependent, while maximum NO production was found to be 34.82 µmol at a 50 µg/mL fucoidan concentration.

Purification, structural analysis and mechanism of murine macrophage cell activation by sulfated polysaccharides from Cystoseira indica.

Sulfated polysaccharides were isolated and purified from the water extract of Cystoseira indica using DEAE Sepharose Fast Flow column to evaluate their structure and macrophage stimulating capacity. Crude and fractionated polysaccharides, CIF1 and CIF2, were mostly composed of neutral sugars (73.1%-78.6%) with relatively lower amounts of acidic sugars (1.3%-9.0%) and sulfate esters (6.9%-9.7%). The polymer chains of polysaccharides were mainly built of different levels of glucose (2.1%-30.8%), fucose (17.2%-24.4%), mannose (17.8%-20.6%) and galactose (16.7%-17.3%). The weight average molecular weight (Mw) of polysaccharides varied between 573.1 × 103 g/mol to 1146.6 × 103 g/mol. The CIF2 polysaccharide, as the most immunostimulating polysaccharide, remarkably induced the release of nitric oxide and inflammatory cytokines including TNF-α, IL-1ß, IL-6 and IL-10 from RAW264.7 murine macrophage cells through NF-κB and PAMKs transduction signaling pathways via cell surface TLR4. The interconnections of sugars in CIF2 polysaccharide were complex with (1→3)-fucopyranose, (1→2,3,4)-glucopyranose, (→1)-galactopyranose, (→1)-xylopyranose, (1→2)-rhamnopyranose and (1→2,3)-mannopyranose units being the most predominant residues.