Community structure of endophytic bacteria of Sargassum thubergii in the intertidal zone of Qingdao in China.

Endophytic bacteria are one of the symbiotic microbial groups closely related to host algae. However, less research on the endophytic bacteria of marine algae. In this study, the endophytic bacterial community of Sargassum thunbergii was investigated using the culture method and high-throughput sequencing. Thirty-nine endophytic bacterial strains, belonging to two phyla, five genera and sixteen species, were isolated, and Firmicutes, Bacillus and Metabacillus indicus were the dominant taxa at the phylum, genus and species level, respectively. High-throughput sequencing revealed 39 phyla and 574 genera of endophytic bacteria, and the dominant phylum was Proteobacteria, while the dominant genus was Ralstonia. The results also indicated that the endophytic bacteria of S. thunbergii included various groups with nitrogen fixation, salt tolerance, pollutant degradation, and antibacterial properties but also contained some pathogenic bacteria. Additionally, the endophytic bacterial community shared a large number of groups with the epiphytic bacteria and bacteria in the surrounding seawater, but the three groups of samples could be clustered separately. In conclusion, there are a variety of functional endophytic bacteria living in S. thunbergii, and the internal condition of algae is a selective factor for the formation of endophytic bacterial communities. This study enriched the database of endophytic bacteria in marine macroalgae, paving the way for further understanding of the interrelationships between endophytic bacteria, macroalgae, and the environment.

The host sex contributes to the endophytic bacterial community in Sargassum thunbergii and their receptacles.

Endophytic bacteria have a complex coevolutionary relationship with their host macroalgae. Dioecious macroalgae are important producers in marine ecosystems, but there is still a lack of research on how sex influences their endophytic bacteria. In this study, the endophytic bacterial communities in male and female S. thunbergii and their reproductive tissues (receptacles) were compared using culture methods and high-throughput sequencing. The endophytic bacterial communities detected by the two methods were different. Among the 78 isolated strains, the dominant phylum, genus, and species were Bacillota, Alkalihalobacillus, and Alkalihalobacillus algicola, respectively, in the algal bodies, while in the receptacles, they were Bacillota, Vibrio, and Vibrio alginolyticus. However, 24 phyla and 349 genera of endophytic bacteria were identified by high-throughput sequencing, and the dominant phylum and genus were Pseudomonadota and Sva0996_ Marine_ Group, respectively, in both the algal body and the receptacles. The two methods showed similar compositions of endophytic bacterial communities between the samples of different sexes, but the relative abundances of dominant and specific taxa were different. The high-throughput sequencing results showed more clearly that the sex of the host alga had an effect on its endophyte community assembly and a greater effect on the endophytic bacterial community in the receptacles. Moreover, most specific bacteria and predicted functional genes that differed between the samples from the males and females were related to metabolism, suggesting that metabolic differences are the main causes of sex differences in the endophytic bacterial community. Our research is the first to show that host sex contributes to the composition of endophytic bacterial communities in dioecious marine macroalgae. The results enrich the database of endophytic bacteria of dioecious marine macroalgae and pave the way for better understanding the assembly mechanism of the endophytic bacterial community of algae.

In vitro and in vivo immuno-enhancing effect of fucoidan isolated from non-edible brown seaweed Sargassum thunbergii.

Fucoidan has been reported to have various biological activities, such as antioxidant, antitumor and anticoagulant, with various health benefits. However, few studies have been conducted to extract fucoidan from Sargassum thunbergii in terms of its immuno-enhancing activities. This aim of this study was to investigate the immuno-enhancing effect of fucoidan (S3) isolated from Sargassum thunbergii through water extraction and ethanol precipitation in RAW 264.7 macrophages and zebrafish. The results showed that S3 contained a relatively high content of fucose and sulfated polysaccharide. Fourier-transform infrared spectroscopy (FTIR) results show that the characteristic peaks at 845 cm-1 and 1220-1270 cm-1 indicate that S3 contains sulfate groups. In vitro, S3 effectively enhanced nitric oxide (NO) production and phagocytic activity. In addition, the results of the study demonstrated that the secretion of tumor necrosis factor-α, interleukin (IL)-6, IL-1ß, and IL-10 was upregulated by nuclear factor kappa B (NF-κB) signaling pathway in a dose-dependent manner. In vivo, S3 activates zebrafish immune responses by promoting secretion of NO and activating the NF-κB pathway. Overall, these results suggest that S3 could be used as a functional ingredient added to nutritional supplements and functional foods.

Effects of instantaneous changes in temperature, light, and salinity on the dynamics of dissolved organic carbon release by Sargassum thunbergii.

Although macroalgae are regarded as the emerging fourth category of "blue carbon", few studies are available on the dynamics of dissolved organic carbon (DOC) release. Sargassum thunbergii is a typical intertidal macroalgae; and tidal action usually triggers instantaneous changes in temperature, light and salinity. Therefore, we investigated the mechanism of short-term changes in temperature, light and salinity on DOC release by S. thunbergii. As well as these factors coupled with desiccation, the combined effect of DOC release was revealed. The results showed the DOC release rate of S. thunbergii was from 0.028 to 0.037 mg C g -1(FW) h-1 under different photosynthetically active radiation (PAR, 0-1500 µmol photons m-2 s-1). The DOC release rate of S. thunbergii was from 0.008 to 0.208 mg C g -1(FW) h-1 under different salinity (5-40). The DOC release rate of S. thunbergii was from 0.031 to 0.034 mg C g -1(FW) h-1 under different temperature (10-30 °C). Either the increase in intracellular organic matter concentration due to increased photosynthesis (change in PAR and temperature, active), cell dehydration due to dry-out process (passive) or the decrease in extracellular salt concentration (passive) would lead to an increase in the difference in osmotic pressure and promote DOC release.

Fucoidan from Sargassum thunbergii obtained via step gradient ethanol precipitation indicate potential anti-obesity and anti-hepatic steatosis in vitro 3T3-L1 and HepG2 cells and in vivo high-fat diet-induced obesity mice.

This study investigated the potential lipid inhibitory and anti-obesity effects of compounds derived from Sargassum thunbergii in vitro and in vivo. We prepared a Celluclast-assisted hydrolysate from Sargassum thunbergii (STC) and three fractional ethanol precipitates (STCF1, STCF2, STCF3). We investigated their proximate composition, and anti-obesity effects in vitro and in vivo. STC and STCFs all significantly reduced intracellular lipid accumulation in PA-treated 3T3-L1 and HepG2 cells. STC, STCF1, and STCF3 had profound anti-obesity effects on high fat diet (HFD)-fed obesity model mice. Oral administration of STC, STCF1, and STCF3 significantly reduced body weight and white adipose tissue (WAT) mass. Furthermore, serum lipid levels were significantly decreased. Additionally, adipose specific hormone levels (adiponectin and fibroblast growth factor-21 (FGF-21)) were significantly decreased, and serum insulin levels were also decreased by STC, STCF1, and STCF3 treatment. A mechanistic study revealed that the adipogenesis and lipolysis associated proteins in epididymal adipose tissue, and free fatty acid oxidation in liver tissues were effectively regulated by STC, STCF1, and STCF3. Overall, our findings show the potent anti-obesity effects of STC, STCF1, and STCF3, achieved by regulation of adipogenesis, lipolysis, and the fatty acid oxidation pathway in HFD-treated obesity model mice.

The effects of intraspecific variation on forecasts of species range shifts under climate change.

As global climate change is altering the distribution range of macroalgae across the globe, it is critical to assess its impact on species range shifts to inform the biodiversity conservation of macroalgae. Latitude/environmental gradients could cause intraspecific variability, which may result in distinct responses to climate change. It remains unclear whether geographical variation occurs in the response of species' populations to climate change. We tested this assumption using the brown alga Sargassum thunbergii, a habitat-forming macroalgae encompassing multiple divergent lineages along the Northwest Pacific. Previous studies revealed a distinct lineage of S. thunbergii in rear-edge populations. Given the phylogeographic structure and temperature gradients, we divided these populations into the southern and northern groups. We assessed the physiological responses of the two groups to temperature changes and estimated their niche differences using n-dimensional hypervolumes. A higher photosynthetic rate and antioxidative abilities were detected in the southern group of S. thunbergii than in the northern group. In addition, significant niche differentiation was detected between the two groups, suggesting the possibility for local adaptation. Given these results, we inferred that the southern group (rear-edge populations) may be more resilient to climate change. To examine climate-driven range shifts of S. thunbergii, we constructed species- and lineage-level species distribution models (SDMs). Predictions of both levels showed considerable distribution contracts along the Chinese coasts in the future. For the southern group, the lineage-level model predicted less habitat loss than the species-level model. Our results highlight the importance of considering intraspecific variation in climate change vulnerability assessments for coastal species.

The differing responses of central carbon cycle metabolism in male and female Sargassum thunbergii to ultraviolet-B radiation.

The enhancement of ultraviolet-B radiation (UV-B) radiation reaching the Earth's surface due to ozone layer depletion is an important topic. Macroalgal species growing in the intertidal zone are often directly exposed to UV-B radiation periodically as the tide changes. In order to better understand the response of macroalgae to UV-B stressed condition, we studied the dominant dioecious intertidal macroalgae Sargassum thunbergii. After consecutive UV-B radiation treatments, we used metabonomics models to analyze and compare the maximum photosynthetic electron transport rate (ETRmax), central carbon cycle metabolism (CCCM) gene expression level, CCCM enzymic activities [pyruvate dehydrogenase and citrate synthase (PDH and CS)], and carbon-based metabolite (including pyruvate, soluble sugar, total amino acid, and lipids) content in male and female S. thunbergii. The results showed that under low and high UV-B radiation, the ETRmax values and six targeted CCCM gene expression levels were significantly higher in males than in females. Under high UV-B radiation, only the CS activity was significantly higher in males than in females. There was no significant difference in PDH activity between males and females. The CCCM models constructed using the metabonomics analysis demonstrate that S. thunbergii males and females exhibit obvious gender differences in their responses to UV-B radiation, providing us with a new understanding of the macroalgal gender differences under UV-B radiation, as past investigations always underestimated their diecious characteristics.

Effects of Water Loss Stress under Tidal Effects on the Epiphytic Bacterial Community of Sargassum thunbergii in the Intertidal Zone.

Intertidal macroalgae face periodic water loss and rehydration caused by daily tidal changes. However, the effect of water loss stress on algal epiphytic bacteria has not yet been reported. In this study, the effects of water loss stress on the epiphytic bacteria community of Sargassum thunbergii were analyzed, and the different responses of epiphytic bacteria to water loss stress were compared between male and female algae. The results showed that after water loss stress, the diversity of the epiphytic bacterial community of S. thunbergii first decreased and then increased. Among the dominant taxa, the abundance of Cyanobacteria decreased significantly, whereas the abundance of Portibacter and Aquimarina first increased and then decreased. Additionally, the indicator species and the abundance of predicted functional genes related to carbon, nitrogen, and sulfur metabolism both changed significantly. More importantly, when the epiphytic bacteria were analyzed separately according to the algal sex, the changes in algal epiphytic bacterial community structure and indicator species were more significant, and there were sexual differences. Therefore, it was concluded that water loss stress has a significant effect on the community structure and function of the epiphytic bacteria on S. thunbergii. Meanwhile, the epiphytic bacteria community of two sexes of S. thunbergii differed in the response to water loss stress. IMPORTANCE Periodic water loss caused by the tide is an important environmental factor that is faced by intertidal macroalgae, but the impact of periodic water loss on the epiphytic bacterial communities associated with macroalgae is still unknown. Through this study, we found that the diversity, the relative abundance of dominant taxa, the indicator species, and the abundance of the predicted functional genes in the epiphytic bacteria on S. thunbergii changed with the time of water loss. Moreover, male and female S. thunbergii exhibited different responses to water loss stress. This study not only paves the way for the delineation of the interactions between S. thunbergii and its epiphytic bacteria but also provides new insights for the mechanisms of the adaptation and evolution of macroalgae in the intertidal zone.

Sex plays a role in the construction of epiphytic bacterial communities on the algal bodies and receptacles of Sargassum thunbergii.

The community structures of epiphytic bacteria on the surface of macroalgae are closely related to their host algae, but there is a lack of research on the differences between the epiphytic bacterial communities of male and female algae and their reproductive tissues. In this study, high-throughput sequencing was used to compare epiphytic bacterial community structures on the intertidal macroalgae Sargassum thunbergii and their receptacles between male and female samples. The epiphytic bacteria on the male and female algal bodies and receptacles had similar community structures with a large number of shared bacteria, but the samples clearly clustered separately, and the abundances of dominant taxa, specific bacteria, and indicator species differed, indicating that epiphytic bacterial communities differed significantly between the male and female S. thunbergii and their receptacles. In addition, the abundance of many predicted functional genes was significantly different between epiphytic bacteria on male and female algal bodies and receptacles, especially metabolism-related genes, and the abundances of predicted functional genes of epiphytic bacteria were significantly higher on both types of male samples than on female samples. Our study confirmed that the sex of the host algae influenced the epiphytic bacterial community structures on algae and algal reproductive tissues, and this role was mainly related to the host metabolism. The results reveal the role of host plant sex in the formation of epiphytic bacterial communities. These findings are helpful for obtaining an in-depth understanding of the construction mechanism of algae-associated bacteria.

Sargassum thunbergii Extract Attenuates High-Fat Diet-Induced Obesity in Mice by Modulating AMPK Activation and the Gut Microbiota.

Sargassum thunbergii (Mertens ex Roth) Kuntze (ST) is a brown alga rich in indole-2-carboxaldehyde. This study aimed to investigate the anti-obesity effects of ethanol extract from ST in in vitro and in vivo models. In 3T3-L1 cells, ST extract significantly inhibited lipid accumulation in mature adipocytes while lowering adipogenic genes (C/epba and Pparg) and enhancing metabolic sensors (Ampk, Sirt1), thermogenic genes (Pgc-1a, Ucp1), and proteins (p-AMPK/AMPK and UCP1). During animal investigation, mice were administered a chow diet, a high-fat diet (HF), or an HF diet supplemented with ST extract (at dosages of 150 and 300 mg/kg bw per day) for 8 weeks (n = 10/group). ST extract administration decreased weight gain, white adipose tissue weight, LDL-cholesterol, and serum leptin levels while improving glucose intolerance. In addition, ST extract increased the expression of Ampk and Sirt1 in adipose tissue and in the liver, as well as p-AMPK/AMPK ratio in the liver, compared to HF-fed mice. The abundance of Bacteroides vulgatus and Faecalibacterium prausnitzii in the feces increased in response to ST extract administration, although levels of Romboutsia ilealis decreased compared with those in HF-fed mice. ST extract could prevent obesity in HF-fed mice via the modulation of AMPK activation and gut microbiota composition.

Diversity of epiphytic bacterial communities on male and female Sargassum thunbergii.

The epiphytic bacteria are the most abundant microorganisms on marine macroalga. However, there are few studies on the distribution of these epiphytic bacteria on male and female Sargassum thunbergii. In this study, the composition and diversity of epiphytic bacterial communities on male and female S. thunbergii were investigated by using the traditional culture-based method and 16S rDNA high-throughput sequencing. The results showed that the dominant bacterial phyla and genera were the same on both male and female S. thunbergii. However, there were significant differences in the relative abundance of epiphytic bacteria at the genus level. Furthermore, male and female S. thunbergii had their own indicative species and specific bacteria. In addition, the predicted functions of the epiphytic bacteria mainly differed in transport and metabolism, environmental adaptation and spore development. This study enriches the baseline knowledge of epiphytic bacteria related to dioecious algae and paves the way for further studies of the relationships between epiphytic microbial communities and the sex of algae.

Photoprotective effects of Sargassum thunbergii on ultraviolet B-induced mouse L929 fibroblasts and zebrafish.

BACKGROUND: Chronic exposure to ultraviolet B (UVB) causes a series of adverse skin reactions, such as erythema, sunburn, photoaging, and cancer, by altering signaling pathways related to inflammation, oxidative stress, and DNA damage. Marine algae have abundant amounts and varieties of bioactive compounds that possess antioxidant and anti-inflammatory properties. Thus, the objective of this study was to investigate the photoprotective effects of an ethanol extract of Sargassum thunbergii. METHODS: Sargassum thunbergii phenolic-rich extract (STPE) was prepared, and its activity against UVB damage was evaluated using L929 fibroblast cells and zebrafish. STPE was extracted and purified by 40% ethanol and macroporous resin XDA-7. Reactive oxygen species (ROS) and antioxidant markers, such as superoxide dismutase (SOD), catalase (CAT) activities, and malondialdehyde (MDA) content were analyzed. The effect of STPE on UVB-induced inflammation was determined by inflammatory cytokine gene and protein expression. The expression of signaling molecules in the Nuclear Factor KappaB (NF-κB) pathway was determined by western blotting. DNA condensation was analyzed and visualized by Hoechst 33342 staining. In vivo evaluation was performed by tail fin area and ROS measurement using the zebrafish model. RESULTS: The total polyphenol content of STPE was 72%. STPE reduced ROS content in L929 cells, improved SOD and CAT activities, and significantly reduced MDA content, thereby effectively alleviating UVB radiation-induced oxidative damage. STPE inhibited the mRNA and protein expression of TNF-α, IL-6, and IL-1α. STPE reversed DNA condensation at concentrations of 20 and 40 µg/mL compared with the UVB control. Moreover, STPE inhibited NF-κB signaling pathway activation and alleviated DNA agglutination in L929 cells after UVB irradiation. Additionally, 1.67 µg/mL STPE significantly increased the tail fin area in zebrafish, and 0.8-1.6 µg/mL STPE effectively eliminated excessive ROS after UVB radiation. CONCLUSIONS: STPE inhibited UVB-induced oxidative stress, inflammatory cytokine expression, and DNA condensation via the downregulation of the NF-κB signaling pathway, suggesting that it prevents UVB-induced photodamage, and has potential for clinical development for skin disease treatment.

Physiological Responses and Metabonomics Analysis of Male and Female Sargassum thunbergii Macroalgae Exposed to Ultraviolet-B Stress.

Ultraviolet-B (UV-B) radiation is a major environmental stress that suppresses or activates defense responses in organisms. UV-B radiation affecting growth and development in intertidal species have been researched for a long time, but a series of unknown knowledge remain in the male and female macroalgae comparison. To compare the different responses of male and female Sargassum thunbergii macroalgae under UV-B radiation, PSII photochemical efficiency determination, metabolomic analysis, and main carbon-based metabolites (including soluble sugar, total amino acid, and lipid) content measuring have been performed in our experiments. Results showed that males have significantly superiority performance in the chlorophyll fluorescence parameters of F v/F m, Y(II), and Y(NO) either low or high UV-B radiation treatments. Metabolomics analysis revealed that carbon and nitrogen metabolism pathways in male and female S. thunbergii were significant components responding to enhanced UV-B radiation. Based on measuring, female S. thunbergii lipid content expressed higher than males without any stimulation. Additionally, under low UV-B radiation stimulation, females total amino acid content shown significantly higher than control group and their lipid content also significantly higher than males. Under high UV-B radiation, males soluble sugar, total amino acid, and lipid content significantly varied from females, which meant that enhancing UV-B stress might altered mainly carbon-based metabolites flowing directions. The present study elucidated the potential role of enhanced UV-B radiation in regulating macroalgae physiological responses, metabolites changing, and reflecting differences between male and female S. thunbergii, contributing to understanding of brown-macroalgae diecious adopting mechanisms in defending intertidal UV-B stresses.

Sulphated glucuronomannan tetramer and hexamer from Sargassum thunbergii exhibit anti-human cytomegalovirus activity by blocking viral entry.

Human cytomegalovirus (HCMV) remains a major public health burden worldwide. The anti-HCMV activity of glucuronomannan oligosaccharides (Gs) and sulphated glucuronomannan oligosaccharides (SGs) was investigated. Among these Gs and SGs, G4S1 and G6S1 (higher sulphated glucuronomannan tetramer and hexamer) showed satisfactory anti-HCMV activity starting at 50 µg/mL and 10 µg/mL, respectively. The results of the morphology, western blotting, qPCR and TCID50 assay showed that they prevented lytic cytopathic changes, inhibited the expression of IE1/2 and UL44, and reduced the UL123 copy number and virus titre significantly. It was interesting to note that degree of sulphation and polymerization was more important for anti-HCMV activity. Moreover, the anti-HCMV activities of G4S1 and G6S1 were stable when stored at 4 °C, -20 °C, and -80 °C for at least three months and mainly occurred in the early stage of HCMV infection through the negative charge of the sulphate groups and the interaction between SGs and the host cells.

Climate-induced range shifts shaped the present and threaten the future genetic variability of a marine brown alga in the Northwest Pacific.

Glaciation-induced environmental changes during the last glacial maximum (LGM) have strongly influenced species' distributions and genetic diversity patterns in the northern high latitudes. However, these effects have seldom been assessed on sessile species in the Northwest Pacific. Herein, we chose the brown alga Sargassum thunbergii to test this hypothesis, by comparing present population genetic variability with inferred geographical range shifts from the LGM to the present, estimated with species distribution modelling (SDM). Projections for contrasting scenarios of future climate change were also developed to anticipate genetic diversity losses at regional scales. Results showed that S. thunbergii harbours strikingly rich genetic diversity and multiple divergent lineages in the centre-northern range of its distribution, in contrast with a poorer genetically distinct lineage in the southern range. SDM hindcasted refugial persistence in the southern range during the LGM as well as post-LGM expansion of 18 degrees of latitude northward. Approximate Bayesian computation (ABC) analysis further suggested that the multiple divergent lineages in the centre-northern range limit stem from post-LGM colonization from the southern survived lineage. This suggests divergence due to demographic bottlenecks during range expansion and massive genetic diversity loss during post-LGM contraction in the south. The projected future range of S. thunbergii highlights the threat to unique gene pools that might be lost under global changes.

Halosmysin A, a Novel 14-Membered Macrodiolide Isolated from the Marine-Algae-Derived Fungus Halosphaeriaceae sp.

Halosmysin A, a new 14-membered macrodiolide with an unprecedented skeleton, was isolated from the fungus Halosphaeriaceae sp. OUPS-135D-4, which, in turn, was obtained from the marine algae Sargassum thunbergii. The chemical structure of the macrodiolide was elucidated using 1D and 2D NMR, as well as high resolution fast atom bombardment mass (HRFABMS) spectral analysis. The absolute stereochemistry was determined via chemical derivatization and comparison with a known compound, (6R,11R,12R,14R)-colletodiol. Additionally, halosmysin A was shown to be very potent against murine P388 leukemia, human HL-60 leukemia, and murine L1210 leukemia cell lines, with IC50 values ranging from 2.2 ± 3.1 to 11.7 ± 2.8 µM.

Indole-4-carboxaldehyde Isolated from Seaweed, Sargassum thunbergii, Attenuates Methylglyoxal-Induced Hepatic Inflammation.

Glucose degradation is aberrantly increased in hyperglycemia, which causes various harmful effects on the liver. Glyoxalase-1 (Glo-1) is a ubiquitous cellular enzyme that participates in the detoxification of methylglyoxal (MGO), a cytotoxic byproduct of glycolysis that induces protein modification (advanced glycation end-products, AGEs) and inflammation. Here, we investigated the anti-inflammatory effect of indole-4-carboxaldehyde (ST-I4C), which was isolated from the edible seaweed Sargassum thunbergii, on MGO-induced inflammation in HepG2 cells, a human hepatocyte cell line. ST-I4C attenuated the MGO-induced expression of inflammatory-related genes, such as tumor necrosis factor (TNF)-α and IFN-γ by activating nuclear factor-kappa B (NF-κB) without toxicity in HepG2 cells. In addition, ST-I4C reduced the MGO-induced AGE formation and the expression of the receptor for AGE (RAGE). Interestingly, both the mRNA and protein expression levels of Glo-1 increased following ST-I4C treatment, and the decrease in Glo-1 mRNA expression caused by MGO exposure was rescued by ST-I4C pretreatment. These results suggest that ST-I4C shows anti-inflammatory activity against MGO-induced inflammation in human hepatocytes by preventing an increase in the pro-inflammatory gene expression and AGE formation. Therefore, it represents a potential therapeutic agent for the prevention of hepatic steatosis.

Antioxidant properties of a sulfated polysaccharide isolated from an enzymatic digest of Sargassum thunbergii.

The protective effect of sulfated polysaccharides (PS-4) isolated from Sargassum thunbergii against oxidative stress in cells and zebrafish was investigated. To begin, a variety of enzymatic extraction methods were used, including five carbohydrases and four proteases. Among the enzymatic extracts, the S. thunbergii Celluclast-assisted extract (STC) demonstrated the strongest scavenging activities against hydrogen peroxide (H2O2). Five polysaccharide fraction (PS-1 to 5) were isolated from STC and purified using ion exchange column chromatography. The antioxidant activity of these fractions was then assessed. PS-4 demonstrated the most promising free radical scavenging and antioxidant activity. PS-4 had a higher content of sulfated polysaccharide than STC. Thus, increasing the proportion of sulfated polysaccharides may be attributed to protection against H2O2-mediated cell damage and oxidative stress in cell lines and in zebrafish. These results suggest that PS-4 could be a potent antioxidant agent in functional foods and cosmetics.

Structure Analysis and Anti-Tumor and Anti-Angiogenic Activities of Sulfated Galactofucan Extracted from Sargassum thunbergii.

Sulfated galactofucan (ST-2) was obtained from Sargassum thunbergii. It was then desulfated to obtain ST-2-DS, and autohydrolyzed and precipitated by ethanol to obtain the supernatant (ST-2-S) and precipitate (ST-2-C). ST-2-C was further fractionated by gel chromatography into two fractions, ST-2-H (high molecular weight) and ST-2-L (low molecular weight). Mass spectrometry (MS) of ST-2-DS was performed to elucidate the backbone of ST-2. It was shown that ST-2-DS contained a backbone of alternating galactopyranose residues (Gal)n (n ≤ 3) and fucopyranose residues (Fuc)n. In addition, ST-2-S was also determined by MS to elucidate the branches of ST-2. It was suggested that sulfated fuco-oligomers might be the branches of ST-2. Compared to the NMR spectra of ST-2-H, the spectra of ST-2-L was more recognizable. It was shown that ST-2-L contain a backbone of (Gal)n and (Fuc)n, sulfated mainly at C4 of Fuc, and interspersed with galactose (the linkages were likely to be 1→2 and 1→6). Therefore, ST-2 might contain a backbone of (Gal)n (n ≤ 3) and (Fuc)n. The sulfation pattern was mainly at C4 of fucopyranose and partially at C4 of galactopyranose, and the branches were mainly sulfated fuco-oligomers. Finally, the anti-tumor and anti-angiogenic activities of ST-2 and its derivates were determined. It was shown that the low molecular-weight sulfated galactofucan, with higher fucose content, had better anti-angiogenic and anti-tumor activities.

The structural features of the sulfated heteropolysaccharide (ST-1) from Sargassum thunbergii and its neuroprotective activities.

Polysaccharide (ST) was prepared from Sargassum thunbergii using hot water. Two fractions (ST-1 and ST-2) were prepared using anion exchange chromatography. One desulfated polysaccharide (ST-1-DS) was also prepared. Electrospray ionization mass spectrometry (ESI-MS) performed on ST-1-DS showed that the desulfated polysaccharides contained methyl glycosides of mono-sulfated and di-sulfated galacto-fucooligosaccharides. This result suggested that ST-1 might contain sulfated galactofucan, which consists of a backbone of alternating (Gal)n and (Fuc)n and sulfated randomly on Gal and mainly on C-2 in Fuc. In addition, ST-1 was degraded in 1M sulfuric acid. The solution was centrifuged, and the supernatant was concentrated and precipitated in ethanol to obtain the precipitate (ST-1-P). ST-1-P was then separated using gel chromatography and anion exchange chromatography to obtain the oligomers. ESI-MS spectra of oligomers indicated that ST-1 mostly contained sulfated glucuronomannan and fucoglucuronan. ESI-MS with collision-induced dissociation tandem mass spectrometry (ESI-CID-MS/MS) suggested that glucuronomannan contained alternating 2-linked Man and 4-linked GlcA, while fucoglucuronan contained 4-linked glucuronan with branched Fuc at C-3. Finally, the neuroprotective activities of ST, ST-1, ST-2 and MIX (a mixture of ST-1 and ST-2) were determined. ST showed the most neuroprotective activity, which indicated that ST might be a good candidate for curing neurodegenerative diseases.