Fucose-containing Polysaccharides Research Articles

Abundance and composition of particles and their attached microbiomes along an Atlantic Meridional Transect

Particulate organic matter plays a significant role in the marine carbon cycle. Its sinking exports organic carbon from the surface to deep oceans. Using fractionated filtration, we analysed particles of 3 – 10 µm and >10 µm and their microbiomes in thirty-five stations along a latitudinal transect of the Atlantic Ocean and provide new insights into the composition, community dynamics, and catabolic potential of particle-attached bacteria. Samples were taken during an Atlantic Meridional Transect (AMT22), which traversed six distinctive ocean provinces. Using 16S rRNA amplicon sequencing and fluorescence in situ hybridisation, we could show a strong variation between particle-attached and free-living bacterial communities at each station and across the biogeographical provinces – a dynamic likely driven by chlorophyll a concentrations, temperature, and the oxygen content of the respective biogeographical provinces. Whereas the <3 µm fraction was primarily composed of SAR11, SAR86, Prochlorococcus and Bacteroidetes of the NS9 and NS5 clades, particle-attached communities were dominated by other Bacteroidetes (Polaribacter spp.), diverse Gammaproteobacteria including members of the genera Alteromonas and Vibrio, Alphaproteobacteria, Planctomycetes, OM27 and Verrucomicrobia. In three provinces, we quantified particle abundance and analysed their glycan composition using four lectins targeting fucose, galactose, N-acetylgalactosamine and mannose. Particles were mainly composed of fucose glycans with only a minor abundance of the other glycans, and particle abundance was directly correlated with the chlorophyll a concentrations. Functional analysis of 54 metagenome-assembled genomes retrieved from bacterial communities attached to small particles showed that particle-attached Bacteroidetes, Planctomycetes and Verrucomicrobia displayed key roles in the degradation of sulfated fucose-containing polysaccharides. We also identified gene clusters potentially encoding the utilisation of mannan and laminarin, suggesting an adaptation to the glycan composition of the particles, potentially resulting in niche diversification. Together, our results provide insights into particle-attached bacteria and their ecological strategies in the Atlantic.

Characterization ofl-fucose isomerase fromPaenibacillus rhizosphaeraeto producel-fuculose from hydrolyzed fucoidan and commercial fucose

l-Fuculose is a valuable rare sugar that is used to treat a variety of ailments, including HIV, cancer, Hepatitis B, human lysosomal disease (fucosidosis), and cardio-protective medications. The enzymatic approach for the production of l-fuculose using l-fucose as a substrate would be an advantageous method with a wide range of industrial applications. The objective of this study is the characterization of recombinant l-fucose isomerase from Paenibacillus rhizosphaerae (Pa-LFI) for the production of l-fuculose from an inexpensive and natural source (fucoidan) as well as its comparison with commercial l-fucose (Sigma-Aldrich). Fucoidan, a fucose-containing polysaccharide (FPs), was isolated from Undaria pinnatifida, subsequently hydrolyzed, and characterized before the enzymatic production of l-fuculose. The results elaborate that FPs contain 35.9% of fucose along with other kinds of monosaccharides. The purified Pa-LFI exhibited a single band at 65 kDa and showed it as a hexamer with a native molecular mass of 396 kDa. The highest activity of 104.5 U mg-1 of Pa-LFI was perceived at a temperature of 50°C and pH 6.5 in the presence of 1 mM of Mn2+. The Pa-LFI revealed a melting temperature (Tm) of 75°C and a half-life of 12.6 h at 50°C. It exhibited that Pa-LFI with aldose substrate (l-fucose), has a stronger isomerizing activity, disclosing Km,kcat, and kcat/Km 86.2 mM, 32 831 min-1, and 335 min-1 mM-1, respectively. After reaching equilibrium, Pa-LFI efficiently catalyzed the reaction to convert l-fucose into l-fuculose and the conversion ratios of l-fuculose from 100 g L-1 of FPs and commercial fucose were around 6% (5.6 g L-1) and 30% (30.2 g L-1), respectively. According to the findings of the current study, the Pa-LFI will be useful in the manufacturing of l-fuculose using an effective and easy approach that produces no by-products.

Effects of Fucoidan Powder Combined with Mineral Trioxide Aggregate as a Direct Pulp-Capping Material.

The development of direct pulp-capping materials with favorable biological and structural properties is an important goal in restorative dentistry. Fucoidan is a sulfated, fucose-containing polysaccharide obtained from brown seaweed, with a wide range of applications; however, its use as a direct pulp-capping material has not been examined. This study aimed to evaluate the mechanical, physical, and biological effects of fucoidan combined with conventional mineral trioxide aggregate (MTA) for direct pulp capping. The capping materials were created using Portland cement (80 wt%) and zirconium oxide (20 wt%) as base components, compared with base components plus 5 wt% fucoidan (PZF5) and base components plus 10 wt% fucoidan (PZF10). The initial and final setting time, compressive strength, chemical components, cell viability, adhesion, migration, osteogenesis, and gene expression were analyzed. Fucoidan significantly reduced the initial and final setting time, regardless of quantity. However, the compressive strength was lower for PZF5. Sulfur levels increased with fucoidan. The biological activity improved, especially in the PZF5 group. Cell migration, Alizarin Red S staining, and alkaline phosphatase activity were upregulated in the PZF5 group. Fucoidan is a useful regenerative additive for conventional pulp-capping materials because it reduces the setting time and improves cell migration and osteogenic ability.

Fucose-containing bacterial exopolysaccharides: Sources, biological activities, and food applications

Bacterial exopolysaccharides are high molecular weight polysaccharides that are secreted by a wide range of bacteria, with diverse structures and easy preparation. Fucose, fucose-containing oligosaccharides (FCOs), and fucose-containing polysaccharides (FCPs) have important applications in the food and medicine fields, including applications in products for removing Helicobacter pylori and infant formula powder. Fucose-containing bacterial exopolysaccharide (FcEPS) is a prospective source of fucose, FCOs, and FCPs. This review systematically summarizes the common sources and applications of FCPs and FCOs and the bacterial strains capable of producing FcEPS reported in recent years. The repeated-unit structures, synthesis pathways, and factors affecting the production of FcEPS are reviewed, as well as the degradation methods of FcEPS for preparing FCOs. Finally, the bioactivities of FcEPS, including anti-oxidant, prebiotic, anti-cancer, anti-inflammatory, anti-viral, and anti-microbial activities, are discussed and may serve as a reference strategy for further applications of FcEPS in the functional food and medicine industries.

Development of a Cryoprotective Formula Based on the Fucose-Containing Polysaccharide FucoPol

This study reports the performance of the polysaccharide FucoPol as an enhancer of cryoprotective formulations. FucoPol at a concentration of 0.25% (w/v) was added to several normothermic Dulbecco-derived solutions and hypothermic Euro-Collins, Custodiol-HTK, and Unisol-CV media, substituting some constituents in the latter class to develop FucoPol-based formulations that were tested for their ability to cryopreserve Vero cells. Supplementation yielded post-thaw cell recovery enhancements of at least 70% and averaged at 82%. The FucoPol-supplemented formulations Dulbecco(+)+FP and Unisol-CVS3 achieved cell viabilities capable of competing with the commercial cryogenic formula CryoStor CS5. Particularly in Unisol-CVS3, mannitol, glucose, gluconate, and dextran were all substituted by 0.25% FucoPol, and still, a similar viability was achieved. Multiparametric correlation clustering showed that FucoPol cryoprotection synergizes best with K+, Ca2+, and Cl- in its microenvironment. Component substitution analysis demonstrated that FucoPol acts consistently as a cryoprotectant, an impermeant, and a colloidal stabilizer, providing a combined alternative to glucose, mannitol, gluconate, and dextran, thus highlighting its potential in the development of high-performing cryoprotective formulas.

Photoprotective effect of the fucose-containing polysaccharide FucoPol

This study reports the performance of FucoPol, a fucose-containing bacterial polysaccharide, as a photostable agent with high absorption yield at concentrations as low as 0.02 % (w/v). FucoPol is non-cytotoxic, efficiently protects from UVA and UVB at concentrations of 0.02–2 % (w/v) and 0.2–2 % (w/v), respectively, has over 94 % overall photostability up to 1.5 h of irradiation time. The lowest concentration studied (0.02 %, w/v) shows a sun protection factor (SPF) of 2.61 ± 0.08. The SPF-to-concentration unit ratio showed that FucoPol is about 60-fold more photoprotective than combinations of common organic and inorganic UV filters. In vitro radiation exposure experiments of adhered Vero epithelial and PM1 keratinocytic cells in the presence of 0.25 % (w/v) FucoPol further showed that cell viability was preserved, and delayed radiation-induced cell death was prevented. Overall, FucoPol outperforms common cosmetic biopolymers like xanthan and fucogel. These results are very promising for the development of bio-based sunscreen formulations.

Silver nanocomposites based on the bacterial fucose-rich polysaccharide secreted by Enterobacter A47 for wound dressing applications: Synthesis, characterization and in vitro bioactivity

This study demonstrates the potential of a high molecular weight fucose-containing polysaccharide secreted by the bacterium Enterobacter A47, named FucoPol, and its silver nanocomposite as potential bioactive materials for wound dressings applications. A green, simple, light-assisted method was used for the synthesis of silver nanoparticles (AgNP) using FucoPol, as stabilizing and reducing agent. The synthesized nanoparticles were spherical, and the main population had a particle size in number ranging between 13 and 30 nm for percentiles 50 and 90, respectively. FucoPol, as well as the functionalized material, besides having no cytotoxicity towards human skin keratinocytes and mouse fibroblasts, also promoted in vitro keratinocytes migration. These observations not only show the safety of FucoPol and FucoPol/AgNP biocomposite, but also their wound healing ability. Moreover, the biocomposite had a strong antimicrobial activity against Staphylococcus aureus ATCC 6538 and Klebsiella pneumoniae CECT 8453, two representative strains of known skin commensal pathogens. These findings demonstrate for the first time the potential of FucoPol for the development of wound healing formulations. Additionally, the FucoPol/AgNP biocomposite might find use in antimicrobial biomaterials, including antibacterial wound healing formulations, which further strengthens the establishment of FucoPol as a bioactive biopolymer.

EXTRACTION AND STRUCTURE ELUCIDATION OF FUCOIDAN FROM MARINE SEAWEED PADINA TETRASTROMATICA HAUCK (PHAEOPHYCEAE)

Seaweeds or the marine macroalgae are one of God’s greatest gift to us through Mother Ocean. The members of Phaeophyceae are exclusively marine and are known for their wide range of diversity and they are rich in cell wall polysaccharide fucoidan and alginate. Fucoidan is a general term for all the fucose-containing polysaccharides from brown seaweeds, while ‘fucan’ will be reserved for the polysaccharide built up with 95% fucose it’s a sulphated polysaccharide. Its highly economical and has abundance applications in the field of pharmaceuticals. Padina tetrastromatica a seaweed abundant in Indian shores was used for the extraction of Fucoidan. Later the structure was elucidated and compared with that of dextran sulphate. This was compared to that of dextran sulphate a standard chemical sulphated polysaccharide while this sulphated polysaccharide is from a natural source.

Demonstration of the cryoprotective properties of the fucose-containing polysaccharide FucoPol

We report the cryoprotective potential of FucoPol, a fucose-containing bacterial exopolysaccharide produced by Enterobacter A47. In vitro cryopreservation assays of Vero, Saos-2, HFFF2 and C2C12 cell lines exposed to a validated non-cytotoxic 2.5 mg/mL FucoPol concentration demonstrated a consistent post-thaw metabolic viability increase. Calorimetric analysis showed a non-colligative, FucoPol concentration-dependent increase of the freezing point (Tf), with minimal change in melting point (Tm). Freezing point variation was corroborated by Polarized Optical Microscopy studies, also showing a reduction of ice crystal dimensions. Its proven shear-thinning behaviour and polyanionicity favour interactivity between the polysaccharide and the water-ice interface, resulting in ice growth inhibition. These findings demonstrate FucoPol’s high promise as a bio-based, biodegradable approach to be implemented into cryopreservation formulations.

Cell wall chemistry and tissue structure underlie shifts in material properties of a perennial kelp

ABSTRACTLaminaria is an abundant kelp genus in temperate nearshore ecosystems that grows with a circannual ‘stop-start’ pattern. Species of Laminaria play important ecological roles in kelp forests worldwide and are harvested commercially as a source of food and valuable extracts. In order to evaluate seasonal differences in tissue properties and composition, we compared the material properties, histology and cell-wall composition of overwintering blades with newly synthesized, actively growing blades from Laminaria setchellii. We found that overwintering blades were fortified with a thicker cortex and increased cell wall investment, leading to increased material strength. Overwintering tissues were composed of higher proportions of cellulose and fucose-containing polysaccharides (i.e. FCSPs, fucoidans) than newly formed blades and were found to possess thicker cell walls, likely to withstand the waves of winter storms. Chemical cell wall profiling revealed that significant proportions of fucose were associated with cellulose, especially in overwintering tissues, confirming the association between cellulose and some fucose-containing polysaccharides. Changes in material properties during the resting phase may allow these kelps to retain their non-growing blades through several months of winter storms. The results of this study demonstrate how one species might regulate its material properties seasonally, and at the same time shed light on the mechanisms that might control the material properties of kelps in general.

Characteristics of fucose-containing polysaccharides from submerged fermentation of Agaricus blazei Murill

Fucose is one of important residues of recognition pattern for many immune cells. In this study, we characterized bioactive fucose-containing acidic polysaccharides from submerged fermentation of Agaricus blazei Murill. We obtained the polysaccharides through a cell-based activity-guided strategy, and used carbohydrate recognition monoclonal antibodies based Enzyme-Linked Immuno Sorbent Assay (ELISA) along with methylation and NMR analyses to investigate the structural characteristics of the polysaccharides. The polysaccharides had Mw of 3.5 × 105 Da. The major sugars were l-fucose, l-arabinose, d-galactose, d-xylose, and d-galacturonic acid in the molar ratio of 6.4, 15.5, 28.5, 14.7, and 25.0% with a small amount of d-glucose, d-mannose, l-rhamnose, and d-glucuronic acid. Results indicated that the bioactive polysaccharides consisted of a (1,4)-Galp and (1,4)-GalAp back bone; (1,2)-Xyl and (1,2)-Rha might also comprise backbone or constitute side chain; linkage (1,5)-Ara and terminal fucosyl residues were also involved in the polysaccharides. Regarding bioactivity, removal of the terminal l-fucosyl residues reduced the TNF-α cytokine stimulating activity of the polysaccharides in a RAW 264.7 macrophage cell-line test, whereas NF-κB and TLR4 affected the polysaccharide-induced TNF-α production.

Fucoidans as a platform for new anticoagulant drugs discovery

AbstractAnionic fucose-containing polysaccharides (fucoidans of brown seaweeds, sulfated fucans and fucosylated chondroitin sulfates of invertebrates) are attracting a rapidly growing research interest due to different types of their biological activity discovered in recent years. In particular, algal fucoidans are characterized by large structural variations depending on the species used for their isolation and by the lack of structural regularity due to random distribution of both carbohydrate and non-carbohydrate substituents along the polymer chains. These features make it difficult to find distinct correlations between structural elements and biological properties of polysaccharides. Nevertheless, there is expectation that systematic structural and biochemical studies of fucoidans will form a basis for the development of new drugs. Herewith we summarize our recent results on the influence of fucoidan structure on blood coagulation.

Immunization of fucose-containing polysaccharides from Reishi mushroom induces antibodies to tumor-associated Globo H-series epitopes

Carbohydrate-based vaccines have shown therapeutic efficacy for infectious disease and cancer. The mushroom Ganoderma lucidum (Reishi) containing complex polysaccharides has been used as antitumor supplement, but the mechanism of immune response has rarely been studied. Here, we show that the mice immunized with a l-fucose (Fuc)-enriched Reishi polysaccharide fraction (designated as FMS) induce antibodies against murine Lewis lung carcinoma cells, with increased antibody-mediated cytotoxicity and reduced production of tumor-associated inflammatory mediators (in particular, monocyte chemoattractant protein-1). The mice showed a significant increase in the peritoneal B1 B-cell population, suggesting FMS-mediated anti-glycan IgM production. Furthermore, the glycan microarray analysis of FMS-induced antisera displayed a high specificity toward tumor-associated glycans, with the antigenic structure located in the nonreducing termini (i.e., Fucα1-2Galβ1-3GalNAc-R, where Gal, GalNAc, and R represent, respectively, D-galactose, D-N-acetyl galactosamine, and reducing end), typically found in Globo H and related tumor antigens. The composition of FMS contains mainly the backbone of 1,4-mannan and 1,6-α-galactan and through the Fucα1-2Gal, Fucα1-3/4Man, Fucα1-4Xyl, and Fucα1-2Fuc linkages (where Man and Xyl represent d-mannose and d-xylose, respectively), underlying the molecular basis of the FMS-induced IgM antibodies against tumor-specific glycans.

Ganoderma lucidum Polysaccharides Induce Macrophage‐Like Differentiation in Human Leukemia THP‐1 Cells via Caspase and p53 Activation

Differentiation therapy by induction of tumor cells is an important method in the treatment of hematological cancers such as leukemia. Tumor cell differentiation ends cancer cells' immortality, thus stopping cell growth and proliferation. In our previous study, we found that fucose-containing polysaccharide fraction F3 extracted from Ganoderma lucidum can bring about cytokine secretion and cell death in human leukemia THP-1 cells. This prompted us to further investigate on how F3 induces the differentiation in human leukemia cells. We integrated time-course microarray analysis and network modeling to study the F3-induced effects on THP-1 cells. In addition, we determined the differentiation effect using Liu's staining, nitroblue tetrazolium (NBT) reduction assay, flow cytometer, western blotting and Q-PCR. We also examined the modulation and regulation by F3 during the differentiation process. Dynamic gene expression profiles showed that cell differentiation was induced in F3-treated THP-1 cells. Furthermore, F3-treated THP-1 cells exhibited enhanced macrophage differentiation, as demonstrated by changes in cell adherence, cell cycle arrest, NBT reduction and expression of differentiation markers including CD11b, CD14, CD68, matrix metalloproteinase-9 and myeloperoxidase. In addition, caspase cleavage and p53 activation were found to be significantly enhanced in F3-treated THP-1 cells. We unraveled the role of caspases and p53 in F3-induced THP-1 cells differentiation into macrophages. Our results provide a molecular explanation for the differentiation effect of F3 on human leukemia THP-1 cells and offer a prospect for a potential leukemia differentiation therapy.

New Microbial Fucose-Containing Polysaccharide from Glycerol byproduct: Rheological Studies

New Microbial Fucose-Containing Polysaccharide from Glycerol byproduct: Rheological Studies

Exploring the potential of biopharmaceutical production by Rigidoporus ulmarius: Cultivation, chemistry, and bioactivity studies

Rigidoporus ulmarius is used as a medicinal fungus in Asia. Three isolates (denoted #61, #62, and #63) of R. ulmarius were collected, and their biological activities were evaluated. Extracted polysaccharides from isolate #63 showed greater inhibition activity compared to isolates #61 and #62 in an in vitro endothelial cell tube formation assay, a standard evaluation of angiogenesis. The polysaccharides and ethanolic extract of isolate #63 dose-dependently suppressed the production of the interferon (IFN)-γ-induced inflammation marker, IP-10. Chemical analyses of the polysaccharides revealed that isolate #63 contained the highest value of fucose at a concentration of 59.1 ± 1.2 μmol/g polysaccharide. These results suggest that fucose-containing polysaccharides may play a role in the inhibitory effect. Isolate #63 showed the highest values of ADP among the three isolates in the ethanolic extract. These results suggest that different isolates from R. ulmarius exhibit different abilities to regulate antiangiogenic and anti-inflammatory processes.

Biological activities of fucose-containing polysaccharide ascophyllan isolated from the brown alga Ascophyllum nodosum.

A fucose-containing, sulfated polysaccharide ascophyllan was isolated from the brown alga Ascophyllum nodosum. Composition analysis demonstrated that ascophyllan mainly contains uronic acid, xylose, fucose, and sulfate half ester in approximately equimolecular proportions, which are evidently distinct from those of alginate and fucoidan. Ascophyllan inhibited the proliferation of U937 cells in a concentration-dependent manner, and DNA-fragmentation and typical apoptotic nuclear morphological changes were observed in the ascophyllan-treated cells. Furthermore, ascophyllan induced the secretion of tumor necrosis factor-alpha (TNF-alpha) and granulocyte colony-stimulating factor (G-CSF) from mouse macrophage cell line RAW264.7.

Inhibitory Effects of Fucoidan in 3T3-L1 Adipocyte Differentiation

Fucoidan is a group of sulfated fucose-containing polysaccharides that derived from non-mammalian origin such as marine brown algae, the jelly coat from sea urchin eggs, and the sea cucumber body wall. However, potential biological activities against obesity from fucoidan were not reported in the literature. The objective of this study was to evaluate protective effect of fucoidan in 3T3-L1 adipocyte differentiation. Preadipocyte 3T3-L1 was treated with 100 and 200 microg/ml fucoidan during adipogenesis. Adipogenesis was determined through Oil Red O staining method and the expression of adipogenic genes aP2, ACC, and PPARgamma. Adipogenesis of 3T3-L1 treated with 100 and 200 microg/ml fucoidan were significantly inhibited at 32.8% and 39.7% using Oil Red O staining method, respectively (P < 0.05). Treating the 3T3-L1 cells with 100 and 200 microg/ml fucoidan significantly decreased the expression of aP2 gene by 6.2% and 27.2%, respectively, of ACC gene by 22.2% and 38.2%, respectively, and of PPARgamma gene by 44.2% and 69.4%, respectively, compared to adipocyte controls (P < 0.05). The results suggest that fucoidan could be used for inhibiting fat accumulation, which is mediated by decreasing aP2, ACC, and PPARgamma gene expression.

A marine strain of flavobacteriaceae utilizes brown seaweed fucoidan.

Fucoidan, a mixture of sulfated fucose-containing polysaccharides, was prepared from Kjellmaniella crassifolia (class Phaeophyceae, order Laminariales, family Laminariaceae) with a yield of about 3.8% dry weight. To isolate enzymes that degrade fucoidan, we first screened marine bacteria for their ability to utilize fucoidan, and isolated one strain of Flavobacteriaceae from seawater that could do this. Phylogenetic analysis of the 16S ribosomal DNA sequence suggested that this strain appeared to belong to a new genus, and was tentatively named Fucobacter marina. The strain utilized L-fucose (17%), D-mannose (91%), D-galactose (46%), and D-glucuronic acid (66%) in the fucoidan from K. crassifolia. The strain partially utilized fucoidan from 2 other seaweeds that belong to the order Laminariales, Undaria pinnatifida (10%) and Lessonia nigrescens (48%).

Dietary fibre and physicochemical properties of edible Spanish seaweeds

Total dietary fibre content of five edible marine Spanish seaweeds: Fucus vesiculosus, Laminaria digitata (Kombu), Undaria pinnatifida (Wakame), Chondrus crispus (Irish moss), and Porphyra tenera (Nori) ranged from 33.6 to 50% of which 19.6 to 64.9% was soluble. For brown algae, the soluble fibre obtained by the AOAC method followed by dialysis, contained uronic acids from alginates and neutral sugars from sulphated fucoidan and laminarin. For red seaweeds, the main neutral sugars corresponded to sulphated galactans: carrageenan (Chondrus) or agar (Nori), respectively. Insoluble fibres (12–40%) were essentially made of cellulose, plus residual fucose-containing polysaccharides, except for the red seaweed Nori, which contained an insoluble mannan and xylan. Protein content in powdered algae was higher in red (20.9–29.8%) than in brown seaweeds (6.9–16%), although protein digestibility was apparently low as inferred from preliminary in vitro results with Fucus and Laminaria. Ashes (21–39.8%) and sulphate content (2.8–10.5%) were high in all seaweeds. Minor components were lipids (0.2–2.5%) and extractable polyphenols (0.4%). Regarding the physicochemical properties, oil retention was low, while swelling, water retention, and cation exchange capacity were higher in brown algae, related to their higher uronic acid plus sulphate content.