Insights into cell wall disintegration of Chlorella vulgaris

With their ability of CO2 fixation using sunlight as an energy source, algae and especially microalgae are moving into the focus for the production of proteins and other valuable compounds. However, the valorization of algal biomass depends on the effective disruption of the recalcitrant microalgal cell wall. Especially cell walls of Chlorella species proved to be very robust. The wall structures that are responsible for this robustness have been studied less so far. Here, we evaluate different common methods to break up the algal cell wall effectively and measure the success by protein and carbohydrate release. Subsequently, we investigate algal cell wall features playing a role in the wall’s recalcitrance towards disruption. Using different mechanical and chemical technologies, alkali catalyzed hydrolysis of the Chlorella vulgaris cells proved to be especially effective in solubilizing up to 56 wt% protein and 14 wt% carbohydrates of the total biomass. The stepwise degradation of C. vulgaris cell walls using a series of chemicals with increasingly strong conditions revealed that each fraction released different ratios of proteins and carbohydrates. A detailed analysis of the monosaccharide composition of the cell wall extracted in each step identified possible factors for the robustness of the cell wall. In particular, the presence of chitin or chitin-like polymers was indicated by glucosamine found in strong alkali extracts. The presence of highly ordered starch or cellulose was indicated by glucose detected in strong acidic extracts. Our results might help to tailor more specific efforts to disrupt Chlorella cell walls and help to valorize microalgae biomass.

Effects and mechanisms of Portulaca oleracea L. polysaccharides on the activation of dendritic cells derived from mice immunized with FMD vaccine

Background Our previous study has showed that Portulaca oleracea L. (POL-P), as an immunoenhancer, could increase the IgG and isotypes antibody titers in mice immunized with foot and mouth disease (FMD) vaccines. However, the structural features and the mechanism of action are still unclear. Enhancing antigen presentation is one of the main ways that immunoenhancer boost immune response. Dendritic cells (DCs) are the most potent antigen presenting cell (APC), which stimulate the initial T cells directly and initiate the specific immune responses. In addition to extracellular factors and intracellular genetic factors, epigenetics plays a major role in the regulation of DCs. In this study, we obtained POL-P, and structural features and monosaccharide composition were analyzed. We evaluated the effect of POL-P on functional maturation of DCs derived from mice immunized with foot and mouth disease (FMD) vaccine and explored the related mechanism responsible for immunoenhancer. The levels of protein and gene related to IL-12p35 and IL-12p40 were determined by western blot and chromatin immunoprecipitation (ChIP) assays. The expressions of TLR2, TLR4 receptors and the downstream molecules of MyD88 and NF-κB were examined using immunohistochemistry. Results The average molecular weight (Mw) of the POL-P was 4×104 Da. The monosaccharide composition of the POL-P was mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and arabinose with a relative mass of 1.2%, 13.2%, 33.5%, 1.2%, 3.3%, 32.2% and 15.4%, respectively. We concluded that co-administration of POL-P with the FMD vaccine could significantly promote DCs maturation of phenotype and the immune function. In addition, the acetylation level of histone H3 of IL-12 was closely connected with the immune activity of DCs. Moreover, POL-P induced immune response was related to up-regulating protein expression of TLR2, TLR4, MyD88 and NF-κB in DCs. Conclusions Our evidence suggested that POL-P could be a potential immunostimulant in the regulation of DCs maturation for FMD vaccine.

Chemical Characterization, Antitumor, and Immune-Enhancing Activities of Polysaccharide from Sargassum pallidum

Searching for natural products with antitumor and immune-enhancing activities is an important aspect of cancer research. Sargassum pallidum is an edible brown alga that has been used in Chinese traditional medicine for the treatment of tumors. However, the purification and application of its active components are still insufficient. In the present study, the polysaccharides from S. pallidum (SPPs) with antitumor and immune-enhancing activities were isolated and purified, and five polysaccharide fractions (SPP-0.3, SPP-0.5, SPP-0.7, SPP-1, and SPP-2) were obtained. The ratio of total saccharides, monosaccharide composition, and sulfated contents was determined, and their structures were analyzed by Fourier transform infrared spectroscopy. Moreover, bioactivity analysis showed that all five fractions had significant antitumor activity against three types of cancer cells (A549, HepG2, and B16), and can induce cancer cell apoptosis. In addition, the results indicated that SPPs can enhance the proliferation of immune cells and improve the expression levels of serum cytokines (IL-6, IL-1β, iNOS, and TNF-α). SPP-0.7 was identified as the most active fraction and selected for further purification, and its physicochemical properties and antitumor mechanism were further analyzed. Transcriptome sequencing result showed that SPP-0.7 can significantly induce the cell apoptosis, cytokine secretion, and cellular stress response process, and inhibit the normal physiological processes of cancer cells. Overall, SPPs and SPP-0.7 may be suitable for use as potential candidate agents for cancer therapy.

Research of Inonotus obliquus Oligosaccharide in Prevention of Hyperlipidemia

In this study, hot water was used to extract Inonotus obliquus oligosaccharide. DEAE-cellulose and Sepharose G-200 were used to purify Inonotus obliquus oligosaccharide. Inonotus obliquus oligosaccharide IOP-2A was obtained. Its molecular weight Mw is about 1000 Da. The monosaccharide composition and molar ratio were glucose : xylose : galactose : mannose = 54.1 : 13.6 : 13.2 : 6.7. In addition, it also contains a small amount of galactose, gluconic acid, rhamnose, and fucose. IOP-2A contained mainly β-glycosidic bonds. Among them, 1,4-glycosidic bonds accounted for 9.2%, and 1,6-glycosidic bonds accounted for 85.1%. Oligosaccharide macromolecules formed a layered structure. Mouse experiments showed that IOP-2A had the function of preventing hyperlipidemia. At the same time, IOP-2A had a certain protective effect on the liver and kidney. The mechanism of IOP-2A in preventing hyperlipidemia was obtained from the perspective of mouse intestinal flora.

Anti-SARS-CoV-2 Activity of Rhamnan Sulfate from Monostroma nitidum

The COVID-19 pandemic is a major human health concern. The pathogen responsible for COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), invades its host through the interaction of its spike (S) protein with a host cell receptor, angiotensin-converting enzyme 2 (ACE2). In addition to ACE2, heparan sulfate (HS) on the surface of host cells also plays a significant role as a co-receptor. Our previous studies demonstrated that sulfated glycans, such as heparin and fucoidans, show anti-COVID-19 activities. In the current study, rhamnan sulfate (RS), a polysaccharide with a rhamnose backbone from a green seaweed, Monostroma nitidum, was evaluated for binding to the S-protein from SARS-CoV-2 and inhibition of viral infectivity in vitro. The structural characteristics of RS were investigated by determining its monosaccharide composition and performing two-dimensional nuclear magnetic resonance. RS inhibition of the interaction of heparin, a highly sulfated HS, with the SARS-CoV-2 spike protein (from wild type and different mutant variants) was studied using surface plasmon resonance (SPR). In competitive binding studies, the IC50 of RS against the S-protein receptor binding domain (RBD) binding to immobilized heparin was 1.6 ng/mL, which is much lower than the IC50 for heparin (~750 ng/mL). RS showed stronger inhibition than heparin on the S-protein RBD or pseudoviral particles binding to immobilized heparin. Finally, in an in vitro cell-based assay, RS showed strong antiviral activities against wild type SARS-CoV-2 and the delta variant.

Purification, Structural Characterization, and Anti-Inflammatory Effects of a Novel Polysaccharide Isolated from Orostachys fimbriata

The present study elucidated the structural characteristics and anti-inflammatory activity of a novel polysaccharide isolated from Orostachys fimbriata, which is a traditional Chinese medicinal plant. O. fimbriata polysaccharide (OFP) was extracted and subsequently purified by chromatography using a DEAE cellulose-52 and Sephadex G-75 column. The molecular weight was determined as 6.2 kDa. HPGPC and monosaccharide composition analysis revealed a homogeneous polysaccharide containing only Glc. Chromatography and spectral analysis showed that the possible chemical structure consisted of →4)-α-Glcp-(1→ and a small quantity of →4,6)-β-Glcp-(1→ in the main chain and →6)-β-Glcp-(1→, α-Glcp-(1→, and β-Glcp-(1→ in the side chain. Morphological analysis using scanning electron microscopy (SEM) and atomic force microscopy (AFM) indicated that OFP had a multi-branched structure, and the sugar chain molecules of polysaccharide appeared aggregated. OFP was found to exhibit anti-inflammatory activity by reducing the secretion of inflammatory factors in RAW264.7 cells and by decreasing the extent of xylene-induced ear swelling in mice.

Structural Characterization of Sulfated Polysaccharide Isolated From Red Algae (Gelidium crinale) and Antioxidant and Anti-Inflammatory Effects in Macrophage Cells

Gelidium crinale, the red algae belonging to Geliaceae Gelidium, is a traditional edible and industrial alga in China. A sulfated polysaccharide (GNP) is successfully separated from Gelidium crinale by acid extraction and two-step column chromatography. Chemical analysis showed that the molecular weight of GNP was 25.8 kDa and the monosaccharide composition had the highest galactose content and confirmed the presence and content (16.5%) of sulfate by Fourier transform infrared spectroscopy (FT-IR) spectrometry as well as barium chloride-gelatin methods. In addition, the effect of GNP on lipopolysaccharide (LPS)-induced oxidative stress and inflammation in macrophages was also evaluated. The research results showed that GNP had fairly strong scavenging activities on 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical, hydroxyl radical, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and had Fe2+-chelating ability in a dose-dependent manner. At the same time, it significantly inhibits the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and the production of pro-inflammatory cytokines in RAW 264.7 cells induced by LPS through blocking the mitogen-activated protein kinase (MAPK)/nuclear factor kappa beta (NF-κB) signaling pathway. These results indicate that GNP may be a latent component anti-inflammation in pharmaceutical and functional food industries.

Evaluation of the Structural, Physicochemical and Functional Properties of Dietary Fiber Extracted from Newhall Navel Orange By-Products

Ultrasound-assisted enzymatic treatment was used to treat Newhall navel orange peel and residue, and then the structural, physicochemical and functional properties of extracted soluble dietary fibers (SDF) and insoluble dietary fibers (IDF) were investigated. The structural properties were determined using scanning electron microscopy, X-ray diffraction, FT-IR and monosaccharide composition. Among these dietary fibers, residue-SDF showed a more complex structure, while peel-IDF exhibited a looser structure. Four samples showed representative infrared spectral features of polysaccharides, typical cellulose crystalline structure and diverse monosaccharide composition. Furthermore, residue-IDF exhibited higher oil-holding capacity (2.08 g/g), water-holding capacity (13.43 g/g) and nitrite ion adsorption capacity (NIAC) than other three samples, and residue-SDF showed the highest swelling capacity (23.33 mL/g), cation exchange capacity (0.89 mmol/g) and cholesterol adsorption capacity (CAC) among these dietary fibers. In summary, this study suggests that the residue-SDF and residue-IDF could be used as the ideal dietary fibers for application in the functional food industry.

Serum N-Glycome Diversity in Teleost and Chondrostrean Fishes

Recent advances in carbohydrate chemistry, chemical biology, and mass spectrometric techniques have opened the door to rapid progress in uncovering the function and diversity of glycan structures associated with human health and disease. These strategies can be equally well applied to advance non-human health care research. To date, the glycomes of only a handful of non-human, non-domesticated vertebrates have been analyzed in depth due to the logistic complications associated with obtaining or handling wild-caught or farm-raised specimens. In contrast, the last 2 decades have seen advances in proteomics, glycoproteomics, and glycomics that have significantly advanced efforts to identify human serum/plasma biomarkers for various diseases. In this study, we investigated N-glycan structural diversity in serum harvested from five cultured fish species. This biofluid is a useful starting point for glycomic analysis because it is rich in glycoproteins, can be acquired in a sustainable fashion, and its contents reflect dynamic physiologic changes in the organism. Sera acquired from two chondrostrean fish species, the Atlantic sturgeon and shortnose sturgeon, and three teleost fish species, the Atlantic salmon, Arctic char, and channel catfish, were delipidated by organic extraction and the resulting protein-rich preparations sequentially treated with trypsin and PNGaseF to generate released N-glycans for structural analysis. Released N-glycans were analyzed as their native or permethylated forms by nanospray ionization mass spectrometry in negative or positive mode. While the basic biosynthetic pathway that initiates the production of glycoprotein glycan core structures is well-conserved across the teleost fish species examined in this study, species-specific structural differences were detected across the five organisms in terms of their monosaccharide composition, sialylation pattern, fucosylation, and degree of O-acetylation. Our methods and results provide new contributions to a growing library of datasets describing fish N-glycomes that can eventually establish species-normative baselines for assessing N-glycosylation dynamics associated with pathogen invasion, environmental stress, and fish immunologic responses.