Sulfated Galactans Research Articles

Non-cytotoxic sulfated agaran from red seaweed Gracilaria cornea induces antitumor phenotype on macrophages in vitro and inhibits tumor growth in vivo

Marine seaweeds are a rich source of sulfated polysaccharides with several biological effects, including antitumor. Some polysaccharides activate macrophages (Mfs) to an antitumor phenotype. In this study, we evaluate whether sulfated galactans (SGs), isolated from red seaweeds Gracilaria cornea (SG-Gc) and Solieria filiformis (SG-Sf), could activate a murine Mfs (RAW 264.7) to an antitumor phenotype. Additionally, we assessed their potential antitumor effects. Both SGs induced nitric oxide release by Mfs. SG-Sf inhibited a murine metastatic melanoma cell line (B16-F10) proliferation compared with the negative control, but SG-Gc did not inhibit B16-F10 proliferation. Notably, a conditioned medium from RAW 264.7 incubated with SG-Gc and SG-Sf inhibited B16-F10 proliferation. Since there was no direct cytotoxicity against B16-F10, we selected SG-Gc for further assays. s SG-Gc induced TNF-α release and increase of the M1 markers iNOS, MHCII, and CD86. Furthermore, 25 mg/kg SG-Gc administered intraperitoneally in B16-F10 melanoma-bearing mice inhibited tumor growth by 60% compared to negative control. In addition, SG-Gc promoted the immunostimulant effect observed on the spleen. No toxic effects were observed in mice treated with SG-Gc. In summary, we identified SG-Gc as an immunomodulatory and antitumor agent.

Red Seaweed (Rhodophyta) Phycocolloids: A Road from the Species to the Industry Application.

Seaweed polysaccharides are versatile both in their functions in seaweed physiology and in their practical applications in society. However, their content and quality vary greatly. This review discusses the main factors that influence the yield and quality of polysaccharides, specifically carrageenans and agars (sulfated galactans) found in red algae species (Rhodophyta). In addition, its historical, current, and emerging applications are also discussed. Carrageenan has been influenced mainly by photosynthetically active radiation (PAR) and nitrogen, while its relationship with temperature has not yet been replicated by recent studies. Agar's seasonal trend has also been found to be more ambiguous than stated before, with light, temperature, nutrients, and pH being influencing factors. In this review, it is also shown that, depending on the compound type, seaweed polysaccharides are influenced by very different key factors, which can be crucial in seaweed aquaculture to promote a high yield and quality of polysaccharides. Additionally, factors like the extraction method and storage of polysaccharides also influence the yield and quality of these compounds. This review also highlights the drawbacks and inadequacy inherent from the conventional (or current) extraction technology approaches.

Bimetallic nanoparticles with sulfated galactan eliminate Vibrio parahaemolyticus in shrimp Penaeus vannamei

Bimetallic (Au/Ag) nanoparticles (BNPs) have shown enhanced antibacterial activity compared to their monometallic counterparts. Sulfated galactans (SG) are a naturally occurring polymer commonly found in red seaweed Gracilaria fisheri. They are biocompatible and biodegradable and environmentally friendly. In this study, we utilized SG in combination with BNPs to develop composite materials that potentially enhance antibacterial activity against shrimp pathogens Vibrio parahaemolyticus and Vibrio harveyi, compared to BNPs or SG alone. BNPs were coated with sulfated galactan (SGBNPs) and characterized using UV–vis spectroscopy, Fourier transform infrared (FTIR) spectroscopy, zeta potential, and transmission electron microscopy (TEM). UV–vis spectroscopy analysis revealed that the surface plasmon peaks of BNPs and SGBNPs appeared at 530 nm and 532 nm, respectively. Zeta potential measurements showed that SGBNPs had a negative charge of −32.4 mV, while the BNPs solution had a positive charge of 38.7 mV. TEM images demonstrated the spherical morphology of both BNPs and SGBNPs with narrow size distributions (3–10 nm). Analysis of the FTIR spectra indicated that SG maintained its backbone structure in SGBNPs, but some functional groups were altered. Notably, SGBNPs showed superior antimicrobial and antibiofilm activities against V. parahaemolyticus and V. harveyi compared to SG and BNPs. Furthermore, treatment with SGBNPs significantly down-regulated the expression of virulence-related genes (toxR, cpsQ, and mfpA) for V. parahaemolyticus 3HP compared to the respective control, bacteria treated with BNPs or SG. Diets supplemented with SGBNPs, BNPs, or SG showed no detrimental impact on the growth of shrimp Penaeus vannamei. Shrimp fed with SGBNPs-supplemented feed showed significantly higher survival rates than those fed with BNPs-supplemented feed when infected with 3HP after being on the supplemented feed for seven days and a subsequent number of fifteen days. These findings collectively demonstrate the benefit of using SG capped Au–Ag BNPs as an antibacterial agent for the prevention and control of Vibrio sp. Infection in shrimp while reducing the risk of environmental contamination.

Antibacterial, anticancer and antioxidant effects of sulfated galactan from Halymenia dilatata: In vitro and in vivo analysis

Seaweed has raised great interest as an excellent source of bioactive components, including vitamins, minerals, polysaccharides, polyphenols, and unique secondary metabolites which reveal broad-spectrum biological activities. In particular, the complex polysaccharides from brown, red and green seaweed possess numerous therapeutic properties. The galactan is routinely used in biomedical research. This study investigates the bioactive properties of sulfated galactan (Hd-SG) derived from the red macroalgae Halymenia dilatata, including its anticancer, antibacterial, anti-biofilm and antioxidant activity. The isolated Hd-SG exhibited potent cytotoxicity against MDA-MB-231 cells with a half-maximum inhibitory concentration (IC50) of 75 μg/ml. It was found that Hd-SG had antibacterial activity against Vibrio alginolyticus, a coastal aquaculture pathogen, with a minimum inhibitory concentration of 300 μg/ml. Toxicology analysis revealed that the isolated Hd-SG was not toxic to zebrafish embryos, even at higher concentrations (100 μg/ml). Moreover, zebrafish embryos treated with Hd-SG at a concentration of 100 μg/ml exhibited a 118.8% reduction in oxidative damage caused by H2O2-induced reactive oxygen species (ROS) production compared to the H2O2 alone treated embryos (562%). In summary, this study demonstrated that the sulfated galactan of H. dilatata has a wide range of therapeutic properties, making it a viable option for biomedical and aquaculture preventive medicine.

Methylation-GC-MS/FID-Based Glycosidic Linkage Analysis of Unfractionated Polysaccharides in Red Seaweeds.

Glycosidic linkage analysis was conducted on the unfractionated polysaccharides in alcohol-insoluble residues (AIRs) prepared from six red seaweeds (Gracilariopsis sp., Prionitis sp., Mastocarpus papillatus, Callophyllis sp., Mazzaella splendens, and Palmaria palmata) using GC-MS/FID analysis of partially methylated alditol acetates (PMAAs). The cell walls of P. palmata primarily contained mixed-linkage xylans and small amounts of sulfated galactans and cellulose. In contrast, the unfractionated polysaccharides of the other five species were rich in galactans displaying diverse 3,6-anhydro-galactose and galactose linkages with varied sulfation patterns. Different levels of cellulose were also observed. This glycosidic linkage method offers advantages for cellulose analysis over traditional monosaccharide analysis that is known for underrepresenting glucose in crystalline cellulose. Relative linkage compositions calculated from GC-MS and GC-FID measurements showed that anhydro sugar linkages generated more responses in the latter detection method. This improved linkage workflow presents a useful tool for studying polysaccharide structural variations across red seaweed species. Furthermore, for the first time, relative linkage compositions from GC-MS and GC-FID measurements, along with normalized FID and total ion current (TIC) chromatograms without peak assignments, were analyzed using principal component analysis (PCA) as a proof-of-concept demonstration of the technique's potential to differentiate various red seaweed species.

Biocatalytic Conversion of Carrageenans for the Production of 3,6-Anhydro-D-galactose.

Marine biomass stands out as a sustainable resource for generating value-added chemicals. In particular, anhydrosugars derived from carrageenans exhibit a variety of biological functions, rendering them highly promising for utilization and cascading in food, cosmetic, and biotechnological applications. However, the limitation of available sulfatases to break down the complex sulfation patterns of carrageenans poses a significant limitation for the sustainable production of valuable bioproducts from red algae. In this study, we screened several carrageenolytic polysaccharide utilization loci for novel sulfatase activities to assist the efficient conversion of a variety of sulfated galactans into the target product 3,6-anhydro-D-galactose. Inspired by the carrageenolytic pathways in marine heterotrophic bacteria, we systematically combined these novel sulfatases with other carrageenolytic enzymes, facilitating the development of the first enzymatic one-pot biotransformation of ι- and κ-carrageenan to 3,6-anhdyro-D-galactose. We further showed the applicability of this enzymatic bioconversion to a broad series of hybrid carrageenans, rendering this process a promising and sustainable approach for the production of value-added biomolecules from red-algal feedstocks.

Jasmonates and Ethylene Shape Floridoside Synthesis during Carposporogenesis in the Red Seaweed Grateloupia imbricata.

Floridoside is a galactosyl-glycerol compound that acts to supply UDP-galactose and functions as an organic osmolyte in response to salinity in Rhodophyta. Significantly, the UDP-galactose pool is shared for sulfated cell wall galactan synthesis, and, in turn, affected by thallus development alongside carposporogenesis induced by volatile growth regulators, such as ethylene and methyl jasmonate, in the red seaweed Grateloupia imbricata. In this study, we monitored changes in the floridoside reservoir through gene expression controlling both the galactose pool and glyceride pool under different reproductive stages of G. imbricata and we considered changing salinity conditions. Floridoside synthesis was followed by expression analysis of galactose-1-phosphate uridyltransferase (GALT) as UDP-galactose is obtained from UDP-glucose and glucose-1P, and through α-galactosidase gene expression as degradation of floridoside occurs through the cleavage of galactosyl residues. Meanwhile, glycerol 3-phosphate is connected with the galactoglyceride biosynthetic pathway by glycerol 3-phosphate dehydrogenase (G3PD), monogalactosyl diacylglyceride synthase (MGDGS), and digalactosyl diacylglyceride synthase (DGDGS). The results of our study confirm that low GALT transcripts are correlated with thalli softness to locate reproductive structures, as well as constricting the synthesis of UDP-hexoses for galactan backbone synthesis in the presence of two volatile regulators and methionine. Meanwhile, α-galactosidase modulates expression according to cystocarp maturation, and we found high transcripts in late development stages, as occurred in the presence of methyljasmonate, compared to early stages in ethylene. Regarding the acylglyceride pool, the upregulation of G3PD, MGDGS, and DGDGS gene expression in G. imbricata treated with MEJA supports lipid remodeling, as high levels of transcripts for MGDGS and DGDGS provide membrane stability during late development stages of cystocarps. Similar behavior is assumed in three naturally collected thalli development stages-namely, fertile, fertilized, and fertile-under 65 psu salinity conditions. Low transcripts for α-galactosidase and high for G3PD are reported in infertile and fertilized thalli, which is the opposite to high transcripts for α-galactosidase and low for G3PD encountered in fertile thalli within visible cystocarps compared to each of their corresponding stages in 35 psu. No significant changes are reported for MGDGS and DGDGS. It is concluded that cystocarp and thallus development stages affect galactose and glycerides pools with interwoven effects on cell wall polysaccharides.

Structure and Binding Properties to Blood Co-Factors of the Least Sulfated Galactan Found in the Cell Wall of the Red Alga Botryocladia occidentalis.

Three different populations of sulfated polysaccharides can be found in the cell wall of the red alga Botryocladia occidentalis. In a previous work, the structures of the two more sulfated polysaccharides were revised. In this work, NMR-based structural analysis was performed on the least sulfated polysaccharide and its chemically modified derivatives. Results have revealed the presence of both 4-linked α- and 3-linked β-galactose units having the following chemical features: more than half of the total galactose units are not sulfated, the α-units occur primarily as 3,6-anhydrogalactose units either 2-O-methylated or 2-O-sulfated, and the β-galactose units can be 4-O-sulfated or 2,4-O-disulfated. SPR-based results indicated weaker binding of the least sulfated galactan to thrombin, factor Xa, and antithrombin, but stronger binding to heparin cofactor II than unfractionated heparin. This report together with our previous publication completes the structural characterization of the three polysaccharides found in the cell wall of the red alga B. occidentalis and correlates the impact of their composing chemical groups with the levels of interaction with the blood co-factors.

Flavivirga spongiicola sp. nov. and Flavivirga abyssicola sp. nov., Isolated from Marine Environments.

Two novel Gram-stain-negative, strictly-aerobic, rod-shaped (1.2 ± 3.4μm × 0.3 ± 0.7μm), and non-motile marine bacterial species, designated MEBiC05379T and MEBiC07777T, were isolated from a marine sponge Pseudaxinella sp. in Gangneung City and deep-sea sediments of the Ulleung basin in the East Sea of Korea, respectively. The 16S rRNA gene sequence analysis revealed high levels of similarities between these strains and members of the genus Flavivirga (97.0-98.4% sequence identities). Both novel strains revealed as mesophilic, neutrophilic in pH and slightly halophilic. Similar to those of other Flavivirga members, the primary cellular fatty acids of both strains were iso-C15:0, iso-C15:1 G, iso-C15:03-OH, and iso-C17:0 3-OH, with MEBiC05379T and MEBiC07777T containing relatively higher proportions of C12:0 and summed feature 3 (C16:1ω7c and/or C16:1ω6c). In both taxa, the major isoprenoid quinone was MK-6. The DNA G + C contents of MEBiC05379T and MEBiC07777T genomes were 32.62 and 32.46mol%, respectively. Compared to other members of Flavivirga, both strains exhibited similar DNA G + C ratio and fatty acids pattern, yet enzyme expression and carbon sources utilization pattern were different. Genomes of the genus Flavivirga showed enzyme preferences to fucoidan and sulfated galactans. Considering the monophyly rule, AAI values delineate the genus Flavivirga from adjacent genera calculated to be 76.0-78.7%. Based on the phenotypic, genomic and biochemical data, strains for MEBiC05379T and MEBiC07777T thus represent two novel species in the genus Flavivirga, for which the names Flavivirga spongiicola sp. nov. (MEBiC05379T [= KCTC 92527T = JCM 16662T]), and Flavivirga abyssicola sp. nov. (MEBiC07777T [= KCTC 92563T = JCM 36477T]) are proposed.

Chemically modified galactans of Grateloupia indica: From production to in vitro antiviral activity

Herpes simplex viruses (HSVs) have an affinity for heparan sulfate proteoglycans on cell surfaces, which is a determinant for virus entry. Herein, several sulfated galactans that mimic the active domain of the entry receptor were employed to prevent HSV infection. They were produced from Grateloupia indica using chlorosulfonic acid-pyridine (ClSO3H.Py)/N,N-dimethylformamide reagent (fraction G-402), SO3.Py/DMF reagent (G-403), or by aqueous extraction (G-401). These galactans contained varied molecular masses (33–55 kDa), and sulfate contents (12–20 %), and have different antiviral activities. Especially, the galactan (G-402) generated by using ClSO3H.Py/DMF, a novel reagent, exhibited the highest level of antiviral activity (EC50 = 0.36 μg/mL) compared to G-403 (EC50 = 15.6 μg/mL) and G-401 (EC50 = 17.9 μg/mL). This most active sulfated galactan possessed a linear chain containing β-(1 → 3)- and α-(1 → 4)-linked Galp units with sulfate group at the O-2/4/6 and O-2/3/6 positions, respectively. The HSV-1 and HSV-2 strains were specifically inhibited by this novel 33 ± 15 kDa galactan, which also blocked the virus from entering the host cell. These results highlight the significant potential of this sulfated galactan for antiviral research and drug development. Additionally, the reagent used for the effective conversion of galactan hydroxy groups to sulfate during extraction may also be useful for the chemical transformation of other natural products.

Sulfated Galactans from Agarophytes: Review of Extraction Methods, Structural Features, and Biological Activities.

Agarophytes are important seaweeds of the Rhodophyta type, which have been highly exploited for industrial use as sources of a widely consumed polysaccharide of agar. In addition to that, sulfated galactans (SGs) from agarophytes, which consist of various functional sulfate groups, have attracted the attention of scientists in current studies. SGs possess various biological activities, such as anti-tumor, anticoagulant, anti-inflammatory, antioxidant, anti-obesity, anti-diabetic, anti-microbial, anti-diarrhea, and gut microbiota regulation properties. Meanwhile, the taxonomy, ecological factors, i.e., environmental factors, and harvest period, as well as preparation methods, i.e., the pretreatment, extraction, and purification conditions, have been found to influence the chemical compositions and fine structures of SGs, which have, further, been shown to have an impact on their biological activities. However, the gaps in the knowledge of the properties of SGs due to the above complex factors have hindered their industrial application. The aim of this paper is to collect and systematically review the scientific evidence about SGs and, thus, to pave the way for broader and otherwise valuable industrial applications of agarophytes for human enterprise. In the future, this harvested biomass could be sustainably used not only as a source of agar production but also as natural materials in functional food and pharmaceutical industries.

Enzymatically-derived oligo-carrageenans interact with α-Gal antibodies and Galectin-3

Carrageenans are linear sulfated galactans synthesized in the Gigartinales, Rhodophyceae species with a varied range of biological properties that are of value to the pharmaceutical and cosmetic sectors. It is unknown how the fine structure of carrageenans dictates their capacity to affect molecular and cellular responses important to wound healing, or the ability to mitigate oxidative, hemostatic and inflammatory processes. Here we use specific endo-carrageenases, from the marine bacterium Zobellia galactanivorans, to produce enzymatically defined neo-series oligosaccharides from carrageenans with 3,6-anhydro-D-galactose on the non-reducing end. Further enzymatic modification of the oligosaccharides was done by treating with the 3,6-anhydro-D-galactosidases from the same bacterium which hydrolyze non-reducing end 3,6-anhydro-D-galactose moieties from neo-carrageenan oligosaccharides. Using the enzymatically produced oligosaccharides, we demonstrate binding to natural human serum antibodies and a monoclonal anti-αGal Ab (m86). The significant interactions with the Galα(1,3)Gal reactive antibodies produced by humans makes them potential potent inducers of complement-dependent reactions and attractive for therapeutic applications. We also demonstrate modulation of the galectin selectivity for the Gal-3 Carbohydrate Recognition Domain (CRD) relative to Gal-1 which has implications to targeting specific biological pathways regulated by the galectins.

Octanoyl esterification of low molecular weight sulfated galactan enhances the cellular uptake and collagen expression in fibroblast cells.

Low molecular weight sulfated galactan (LMSG) supplemented with octanoyl ester (Oct-LMSG) demonstrated superior wound healing activity compared to the unsupplemented LMSG in a fibroblast wound model. To test the hypothesis that the increased bioactivity of Oct-LMSG may depend on its penetration into the plasma membrane, its cellular uptake was investigated and collagen production in fibroblast cells was assessed for the first time. The cellular uptake of Oct-LMSG was examined using indirect immunofluorescence and a confocal laser scanning microscope. In addition, the degree of fibroblast activation associated with this uptake was evaluated. The results indicated increased LMSG internalization in fibroblasts treated with Oct-LMSG. Transmission electron micrographs revealed the ultrastructure of active protein production in fibroblasts upon treatment with Oct-LMSG. In addition, Oct-LMSG upregulated the expression of type I collagen mRNA and proteins, as well as related signaling molecules involved in collagen synthesis, including collagen type I α1 chain (Col1A1), Col1A2, phosphorylated (p)-Smad2/3 and p-Smad4. The current findings support the notion that the supplementation of LMSG with octanoyl enhanced its cellular uptake into fibroblasts and, as a result, regulated the expression of type I collagen in fibroblasts via the activation of the Smad signaling pathway. This study demonstrates the therapeutic potential of Oct-LMSG in promoting tissue regeneration.

A pyruvylated and sulfated galactan from the green alga Dictyosphaeria cavernosa: Structure, anticoagulant property and inhibitory effect on zebrafish thrombosis

A pyruvylated and sulfated galactan from the green alga Dictyosphaeria cavernosa, designated PSG, was obtained by dilute alkali extraction, ion-exchange and gel filtration chromatography. The backbone of PSG was composed of 3-linked β-d-Galp units with partial sulfation on C-4 and C-6. Pyruvate ketals were linked to O-3 and O-4 of nonreducing terminal β-d-Galp, as well as O-4 and O-6 of 3-linked β-d-Galp. The branches consisting of 6-linked β-d-Galp(4SO4) and β-d-Galp(3,4-Pyr)-(1→ units were located at C-6 of 3-linked β-d-Galp unit. PSG possessed obvious anticoagulant effect in vitro as assessed by the tests of activated partial thromboplastin time and thrombin time. The assay of anticoagulant mechanism showed that PSG promoted thrombin inactivation mediated by heparin cofactor-II and antithrombin-III (ATIII), and could effectively potentiate factor Xa inactivation by ATIII. The antithrombotic activity of PSG in vivo was assessed by phenylhydrazine (PHZ)-induced zebrafish thrombotic model. The results indicated that PSG obviously reduced peripheral erythrocytes aggregation, enhanced cardiac blood flow and improved peripheral platelet circulation, and PSG possessed a marked inhibitory effect on the PHZ-induced zebrafish thrombosis. Thus, PSG is a hopeful anticoagulant and antithrombotic polysaccharide.

Carrageenan impact on digestive proteolysis of meat proteins in meatballs or soluble hydrolyzed collagen

In a health-conscious age, vivid discussion has been made on the healthfulness of processed foods and food additives. This study focuses on carrageenan (CGN), an approved but debated family of sulphated galactans from algae used as gelling, thickening and stabilizing agents but with indications of possible adverse effects, including as an inhibitor of digestive proteolysis. To challenge this inhibitory hypothesis, food-grade kappa-, iota and lambda-CGN preparations were used to produce beef meatballs whose proteolysis was studied using an in vitro digestion model coupled to various proteomic analyses. Results show that CGN anti-nutritional effects are abolished in beef meatballs. Specifically, proteomic analysis of gastric digesta of myosin light chain 1 (MYL1), alpha skeletal muscle (ACTA1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and fructose-bisphosphate aldolase (ALDOA) reveal no appreciable differences in the profiles of bioaccessible peptides. Separate digestions of a soluble collagen hydrolysate show CGN does inhibit proteolysis of soluble collagen, therefore supporting the notion that the meat matrix confers a shielding effect that eliminates CGN ability to interfere with digestive proteolysis. Thus, this work shows that CGN ability to hinder digestive proteolysis may not apply to all foods and contributes evidence important to the discussions on CGN uses, indications and regulatory status.

Complex sulfated galactans from hot water extracts of red seaweed Asparagopsis taxiformis comprise carrageenan and agaran structures

Hot water extraction from the red seaweed Asparagopsis taxiformis yielded three extracts which showed sulfated galactans with a D:L-galactose ratio non consistent with carrageenan or agaran backbones. The major extract was fractionated by cetrimide precipitation and redissolution with increasing sodium chloride concentrations due to their low solubility. Seven fractions were obtained, and studied by methylation analysis, desulfation-methylation, and NMR spectroscopy of the partially hydrolyzed and the native samples. Fractions with the highest yield were those obtained at high concentrations of NaCl. They comprised both agaran and crageenan structures in considerable amounts. The main agaran structures were β-D-galactose 4-sulfate and β-D-galactose 2-sulfate units linked to α-L-galactose 2,3-disulfate residues, and β-D-galactose linked to α-L-galactose 3-sulfate or 6-sulfate, or substituted with single stubs of β-D-xylose on C3, while the carrageenan structures comprised β-D-galactose (2-sulfate) linked to α-D-galactose 3-sulfate or 2,3-disulfate, and β-D-galactose 2,4-disulfate linked to α-D-galactose 2,3-disulfate. Between the less sulfated fractions, the one obtained by solubilization in 0.5 M NaCl was mainly constituted by agarans, which included 3,6-anhydro-α-L-galactose units. Anticoagulant activity was assayed by general coagulation tests (aPTT and TT), showing a moderate action compared with heparin. This is the first detailed study of the sulfated galactans from the order Bonnemaisoniales.

Jasmonates disrupt carrageenan synthesis during carposporogenesis in the red seaweed Grateloupia imbricata

In this study, we monitored changes in cell wall sulfated galactans in the red seaweed Grateloupia imbricata after the rapid (48 h) induction of cystocarp maturation and carpospore development by the volatile hormone methyl jasmonate (MEJA). Synthesis of sulfated galactans, carrageenans, which requires sulfated UDP galactose, was followed by expression analysis of genes encoding phosphoglucomutase (PGM), galactose-1-phosphate uridyltransferase (GALT), and those responsible for sulfate assimilation (S-transporter and sulfate adenylyltransferase, SAT). In addition, the expression of carbohydrate sulfotransferase and galactose-6-sulfurylase responsible for the addition and removal of sulfate groups to the galactans backbone were evaluated. Structural changes, such as thallus softening, were analyzed using Fourier-transformed infrared spectroscopy (FTIR) spectra, and ploidy was assessed using flow cytometry. The results showed downregulation of most of the genes encoding precursors or those in charge of transformation into sulfated hexoses for cell wall synthesis. Furthermore, alterations in the FTIR spectrum of carrageenan from MEJA-induced fertile thalli, such as the disappearance and flattening of absorption bands, were observed. The promptness of these events and the ploidy of thalli and spores seem to confirm the induction of mitotic production of haploid spores by MEJA.

Biopolymer Kappa Carrageenan with Ammonium Chloride as Electrolyte for Potential Application in Organic Battery

Carrageenan is a generic name for a family of natural, water-soluble, sulphated galactans isolated from red seaweeds and exploited commercially. The biopolymer of kappa carrageenan has been known to be used as electrolyte in electrochemical device since it shows good ionic conductivity characteristic. In this study, we attempt to study the chemical, morphology, and electric properties of biopolymer kappa carrageenan. We developed a free-standing film of kappa carrageenan with addition of ammonium chloride as an electrolyte for an organic battery prototype. We prepared the solution by mixing kappa carrageenan, ammonium chloride and water to form a gel with a particular concentration. Then, the gel was coated on the substrate and cured at 50°C for 4 hours. The final free-standing film product reveals a thickness about 100-200 mm as captured by SEM image in cross-section view. The morphology of kappa carrageenan with or without ammonium chloride clearly shows a non-homogeneous surface that attributed to the nature characteristics of kappa carrageenan immiscible. The addition of ammonium chloride into kappa carrageenan forms a smoother surface that show good mixture of kappa carrageenan. FTIR spectra of the samples show the interaction of ammonium chloride to the host polymer of kappa carrageenan as indicated by the shifted of the O-H peak from 3448 to 3446 cm-1 and from 3288 to 3207 cm-1 while the peak of 2924 cm-1 is disappeared after addition of the ammonium chloride. The implementation of this film in an organic C_Zn battery prototype shows that battery’s voltage reached 2.1 Volt by charging. Then, the battery can be used to emit an LED with 20 µA electrical current for about 1 hour in discharging process.

Nuclear magnetic resonance-based structural elucidation of novel marine glycans and derived oligosaccharides.

Marine glycans of defined structures are unique representatives among all kinds of structurally complex glycans endowed with important biological actions. Besides their unique biological properties, these marine sugars also enable advanced structure-activity relationship (SAR) studies given their distinct and defined structures. However, the natural high molecular weights (MWs) of these marine polysaccharides, sometimes even bigger than 100 kDa, pose a problem in many biophysical and analytical studies. Hence, the preparation of low MW oligosaccharides becomes a strategy to overcome the problem. Regardless of the polymeric or oligomeric lengths of these molecules, structural elucidation is mandatory for SAR studies. For this, nuclear magnetic resonance (NMR) spectroscopy plays a pivotal role. Here, we revisit the NMR-based structural elucidation of a series of marine sulfated poly/oligosaccharides discovered in our laboratory within the last 2 years. This set of structures includes the α-glucan extracted from the bivalve Marcia hiantina; the two sulfated galactans extracted from the red alga Botryocladia occidentalis; the fucosylated chondroitin sulfate isolated from the sea cucumber Pentacta pygmaea; the oligosaccharides produced from the fucosylated chondroitin sulfates from this sea cucumber species and from another species, Holothuria floridana; and the sulfated fucan from this later species. Specific 1H and 13C chemical shifts, generated by various 1D and 2D homonuclear and heteronuclear NMR spectra, are exploited as the primary source of information in the structural elucidation of these marine glycans.

Antiviral activity of marine sulfated glycans against pathogenic human coronaviruses

Great interest exists towards the discovery and development of broad-spectrum antivirals. This occurs due to the frequent emergence of new viruses which can also eventually lead to pandemics. A reasonable and efficient strategy to develop new broad-spectrum antivirals relies on targeting a common molecular player of various viruses. Heparan sulfate is a sulfated glycosaminoglycan present on the surface of cells which plays a key role as co-receptor in many virus infections. In previous work, marine sulfated glycans (MSGs) were identified as having antiviral activities. Their mechanism of action relies primarily on competitive inhibition of virion binding to heparan sulfate, preventing virus attachment to the cell surface prior to entry. In the current work we used pseudotyped lentivirus particles to investigate in a comparative fashion the inhibitory properties of five structurally defined MSGs against SARS-CoV-1, SARS-CoV-2, MERS-CoV, and influenza A virus (IAV). MSGs include the disaccharide-repeating sulfated galactan from the red alga Botryocladia occidentalis, the tetrasaccharide-repeating sulfated fucans from the sea urchin Lytechinus variegatus and from the sea cucumber Isostichopus badionotus, and the two marine fucosylated chondroitin sulfates from the sea cucumbers I. badionotus and Pentacta pygmaea. Results indicate specificity of action against SARS-CoV-1 and SARS-CoV-2. Curiously, the MSGs showed decreased inhibitory potencies against MERS-CoV and negligible action against IAV. Among the five MSGs, the two sulfated fucans here studied deserve further attention since they have the lowest anticoagulant effects but still present potent and selective antiviral properties.