sepharose and porphyran

Porphyra is one of the most economically important red algae in the world. The functional components extracted from Porphyra such as porphyrans, proteins, lipids, and minerals have strong physiological activities. Porphyran, a sulfated galactan, is composed of alternating 1,4-linked α-l-galactopyranose-6-sulfate (L6S) and 1,3-linked β-d-galactopyranose (G). Porphyran and oligo-porphyran have a series of pharmacological and biological functions, such as antioxidation, anticancer, antiaging, antiallergic, immunomodulatory, hypoglycaemic, and hypolipidemic effects. Thus, red algae Porphyra-derived porphyran and oligo-porphyran have various potential applications in food, medicine, and cosmetic fields. For better application, this review introduces and summarizes the structure and source of porphyran as well as the preparation methods, biological activities, and potential applications of porphyran and oligo-porphyran. Moreover, the future research directions and emphasis of porphyran and oligo-porphyran preparation as well as their functional activities and applications are highlighted and prospected.

Topics: Antioxidants; Polymerization; Porphyra; Sepharose

Seaweeds are some of the largest producers of biomass in the marine environment and are rich in bioactive compounds that are often used for human and animal health. Porphyran and carrageenan are natural compounds derived from red seaweeds. The former is a characteristic polysaccharide of

Topics: Carrageenan; Humans; Immunity; Neoplasms; Polysaccharides; Seaweed; Sepharose

The hazardous effects of current nanoparticle synthesis methods have steered researchers to focus on the development of newer environmentally friendly and green methods for synthesizing nanoparticles using nontoxic chemicals. The development of environmentally friendly methods of nanoparticle synthesis with different sizes and shapes is one of the pressing challenges for the current nanotechnology. Several novel green approaches for the synthesis of AuNPs have been explored using different natural sources, such as plants, algae, bacteria, and fungi. Among organisms, algae and blue-green algae are of particular interest for nanoparticle synthesis. Gold nanoparticles (AuNPs) have a range of applications in medicine, diagnostics, catalysis, and sensors because of their significant key roles in important fields. AuNPs have attracted a significant interest for use in a variety of applications. The widespread use of AuNPs can be accredited to a combination of optical, physical, and chemical properties as well as the miscellany of size, shape, and surface composition that has been adopted through green synthesis methods.

Topics: Antioxidants; Bacterial Infections; Catalysis; Cell Line, Tumor; Cyanobacteria; Fungi; Gold; Green Chemistry Technology; Humans; Metal Nanoparticles; Nanotechnology; Neoplasms; Plants; Polymers; Seaweed; Sepharose; Surface Properties

Over the past two decades, there have been substantial progress and a growing body of research on using natural polymeric biomaterials in emerging biomedical applications. Among different natural biopolymers, polysaccharides have gained considerable attraction among biomedical scientists and surgeons due to their biocompatibility, biodegradability, anti-inflammatory, and antimicrobial properties. In recent years, algalbased polysaccharides including agar, alginate, and carrageenan, have been broadly suggested for different biomedical applications.. The aim of this paper is discussing various possible applications of algal-based polysaccharides in biomedical engineering particularly in controlled drug delivery systems. The main properties of each algal polysaccharide will be discussed, and particular drug delivery applications will be presented.. Algal polysaccharides can be detected in a group of photosynthetic unite as their key biomass constituents. They provide a range of variety in their size, shape, liquefaction, chemical stability, and crosslinking ability. In addition, algal polysaccharides have shown exceptional gelling properties including stimuli-responsive behavior, softness, and swelling properties.. All the mentioned properties of alga polysaccharides lead to their successful usage in biomedical applications specially targeted and controlled drug delivery systems such as particles, capsules, and gels.

Topics: Agar; Alginates; Biocompatible Materials; Carrageenan; Drug Delivery Systems; Phaeophyceae; Polysaccharides; Rhodophyta; Sepharose

Pyropia (Porphyra), commonly known as nori or laver, is an important food source in many parts of the world. Edible dried Pyropia contains numerous nutrients and biofunctional components, including proteins, vitamins, eicosapentaenoic acid, minerals, carotenoids, mycosporine-like amino acids, and carbohydrate, and one of the compounds which we are interested in is porphyran, a sulfated polysaccharide comprising the hot-water-soluble portion of Pyropia cell walls. Researchers have performed a large number of in-depth studies on the biological activity and potential therapeutic applications of porphyrans and oligoporphyrans.. This mini review aims to provide comprehensive and update overview on the source, extraction, structure, biological activities and structure-activity relationships of porphyrans and oligoporphyrans based on the studies in the past 30 years which were included in Web of Science.. The structure of porphyran has been basically determined given that its straight chain is relatively simple, and the skeleton structure has been described. The extraction methods were simplified continuously, but different extraction methods and post- processing methods still had great influence on the structure and composition of porphyran, so there was no standardized extraction process which can achieve quality control until now. In order to obtain oligoporphyrans, there are a variety of degradation methods, including chemical method, physical method and enzymatic method, but it is worth mentioning that specific degradation enzyme is still unavailable. Studies on the biological and pharmacology properties include antioxidant, anti-tumor, anti-inflammatory, immunomodulation, anti-cardiovascular and cerebrovascular diseases and drug delivery.. Owing to the therapeutic potential and drug delivery applications, porphyran and oligoporphyrans are expected to be further developed as a medicine against human diseases, as well as a supplement in cosmetics and health products.

Topics: Cell Wall; Porphyra; Sepharose; Structure-Activity Relationship

Marine polysaccharides have been found as the principle component in cell wall structures of seaweeds or exoskeletons of crustaceans. Due to numerous pharmaceutical properties of marine polysaccharides such as antioxidant, anti-inflammatory, antiallergic, antitumor, antiobesity, antidiabetes, anticoagulant, antiviral, immunomodulatory, cardioprotective, and antihepatopathy activities, they have been applied in many fields of biomaterials, food, cosmetic, and pharmacology. Recently, several marine polysaccharides such alginate, porphyran, fucoidan, and chitin and its derivatives have been evidenced as downregulators of allergic responses due to enhancement of innate immune system, alteration of Th1/Th2 balance forward to Th1 cells, inhibition of IgE production, and suppression of mast cell degranulation. This contribution, therefore, focuses on antiallergic properties of marine polysaccharides and emphasizes their potential application as bioactive food ingredients as well as nutraceuticals for prevention of allergic disorders.

Topics: Alginates; Anti-Inflammatory Agents; Antioxidants; Antiviral Agents; Chitin; Chitosan; Dietary Supplements; Glucuronic Acid; Hexuronic Acids; Hypersensitivity; Immunoglobulin E; Mast Cells; Oligosaccharides; Polysaccharides; Seaweed; Sepharose; Th1-Th2 Balance

Polysaccharides are macromolecules made up of many monosaccharides joined together by glycosidic bonds. Polysaccharides from marine sources are widely distributed as the principle component in cell wall structures of seaweeds or exoskeletons of crustaceans. So far, marine polysaccharides have been used in many fields of biomaterials, food, cosmetic, and pharmacology. Especially, numerous pharmaceutical properties of marine polysaccharides have been revealed such as antioxidant, anti-inflammatory, antiallergic, antitumor, antiobesity, antidiabetes, anticoagulant, antiviral, immunomodulatory, cardioprotective, antihepatopathy, antiuropathy, and antirenalpathy activities. Recently, several marine polysaccharides such alginate, porphyran, fucoidan, and chitin and its derivatives have been found as modulators of allergic responses due to enhancing innate immune system, altering Th1/Th2 balance, inhibiting IgE production, and suppressing mast cell degranulation. This contribution, therefore, focuses specially on the immunomodulatory effect of marine polysaccharides and emphasizes their potential application as candidates of pharmaceuticals as well as nutraceuticals to prevent allergic disorders.

Topics: Alginates; Animals; Aquatic Organisms; Chitin; Drug Industry; Glucuronic Acid; Hexuronic Acids; Humans; Hypersensitivity; Immunity; Immunologic Factors; Polysaccharides; Sepharose

Porphyran is known to inhibit immune cell function. Previously, porphyran was shown to prevent lipopolysaccharide-induced sepsis in mice. However, studies on the inhibitory effects of porphyran during colitis are currently lacking. In this study, we evaluated the effects of Pyropia yezoensis-derived porphyran on dextran sodium sulfate (DSS)-induced acute and chronic colitis. The oral or intraperitoneal administration of porphyran inhibited the progression of DSS-induced colitis in mice, with the former also preventing immune cell infiltration in the colon. The levels of intracellular interferon-γ and interleukin-17 in T cells decreased when porphyran was administered orally. Porphyran inhibited T cell activation by suppressing dendritic cells (DCs) and macrophages. Porphyran prevented pathogen-associated molecular pattern and damage-associated molecular pattern-dependent DC and macrophage activation. Finally, porphyran attenuated chronic colitis caused via the long-term administration of DSS. These findings indicate that the oral administration of porphyran can inhibit DSS-induced colitis by suppressing DC and macrophage activation.

Topics: Animals; Colitis; Colon; Dendritic Cells; Dextran Sulfate; Disease Models, Animal; Mice; Mice, Inbred C57BL; Rhodophyta; Sepharose

Porphyran, a polysaccharide composed of red algae, is a source of a multifunctional oligosaccharide material and raw biomass with various physiological activities. The glycolysis of porphyrans into oligosaccharides through various porphyranases is an approach for obtaining high-quality and promising alternative resources. In this study, porphyran was extracted from Porphyra yezoensis and used as a research substrate. We also established an efficient hydrolysis method using an enzymatic complex obtained through cohesin-dockerin interactions that degrade natural polysaccharides. The cohesion-dockerin interaction is designed to genetically bind the dockerin module to the end of an existing enzyme and then attach the cohesin module to obtain a protein complex. The designed protein complex has been shown to further increase the activity on the substrate, which can be considered a useful method to obtain efficient oligosaccharides or monosaccharides through hydrolysis of red algae for bioresources.

Topics: Bacterial Proteins; Hydrolysis; Multienzyme Complexes; Multifunctional Enzymes; Sepharose

Porphyran is a promising bioactive polysaccharide majorly composed of 4-linked α-l-galactopyranose-6-sulfate (L6S) and 3-linked β-d-galactopyranose (G) disaccharide repeating units. Carbohydrate-binding modules (CBMs) have been verified to be essential tools for investigating polysaccharides. However, no confirmed CBM binding to porphyran has been hitherto reported. In this study, an unknown domain with a predicted β-sandwich fold from a potential GH86 porphyranase was discovered, and further recombinantly expressed. The CBM protein (named FvCBM99) presented a desired specificity for porphyran tetrasaccharide with an affinity constant of 1.9 × 10

Topics: Galactose; Oligosaccharides; Polysaccharides; Sepharose

Prebiotics, such as algal polysaccharides, can be used to manage metabolic diseases by modulating gut microbiota. However, the effect of Pyropia yezoensis porphyran (PYP), a red algal polysaccharide, on gut microbiota has not been reported. Thus, the objective of this study was to determine effects of PYP on metabolic disorders caused by high sucrose (HS) and underlying mechanisms involved in such effects.. Biochemical analysis demonstrated that an HS diet increased triglyceride and circulating sugar contents (metabolic abnormalities) in Drosophila larvae. It also increased the relative abundance of harmful microbiota within the larvae as identified by 16S ribosomal DNA analysis. PYP supplementation at 25 and 50 g kg. PYP exhibits anti-metabolic disorder activity by modulating gut microbiota, thereby supporting the development of PYP as a functional prebiotic derived from red algae food. Copyright © 2022 John Wiley & Sons, Ltd. © 2022 Society of Chemical Industry.

Topics: Animals; Diet, High-Fat; Drosophila melanogaster; Gastrointestinal Microbiome; Metabolic Diseases; Mice; Mice, Inbred C57BL; Polysaccharides; Prebiotics; Rhodophyta; Sepharose; Sucrose; Triglycerides

Agarase is of vital significance for functional agaro-oligosaccharides production from algal dived agarose. Especially, the exolytic agarases have the advantage of obtaining agaro-oligosaccharides with a specific degree of polymerization. Herein, we cloned and expressed a novel glycoside hydrolase (GH) 50 family β-agarase OUC-PgJC50 from

Topics: Glycoside Hydrolases; Molecular Docking Simulation; Oligosaccharides; Sepharose

Degraded porphyran is a bioactive polysaccharide extracted from Porphyra haitanensis (P. haitanensis). According to the previous studies, it produced anti-inflammatory activity, but little is known about its effects on depression.. As inflammation is one of the critical factors involved in the development of depression, this study aims to elucidate the potential antidepressant-like effects of degraded porphyran. The results show that acute porphyran treatment decreased the immobility time in despair tests. In addition, subchronic porphyran administration reverses depressive-like behaviors in lipopolysaccharide (LPS)-treated mice. Meanwhile, porphyran inhibits NF-κB/NLRP3 signaling, proinflammatory cytokine release, and microglial activation in the hippocampus. Moreover, chronic porphyran treatment activates hippocampal brain derived neurotrophic factor (BDNF)/TrkB/ERK/CREB signaling pathway in chronic unpredictable mild stress (CUMS) in mice. As a result, neurogenesis and spinogenesis are maintained.. The findings of the present study indicate that degraded porphyran intake provides a potential strategy for depression treatment, which is mediated by the inhibition of neuroinflammation and the enhancement of neurogenesis and spinogenesis in the central nervous systems.

Topics: Animals; Antidepressive Agents; Brain-Derived Neurotrophic Factor; Male; Mice; Mice, Inbred C57BL; Microglia; Molecular Docking Simulation; Neurogenesis; Neuroinflammatory Diseases; Porphyra; Sepharose; Toll-Like Receptor 4

Porphyran and its derivatives possess a variety of biological activities, such as ameliorations of oxidative stress, inflammation, hyperlipemia, and immune deficiencies. In this study, we evaluated the potential efficacy of porphyran-derived oligosaccharides from Porphyra yezoensis (PYOs) in alleviating nonalcoholic fatty liver disease (NAFLD) and preliminarily clarified the underlying mechanism. NAFLD was induced by a high-fat diet for six months in C57BL/6J mice, followed by treatment with PYOs (100 or 300 mg/kg/d) for another six weeks. We found that PYOs reduced hepatic oxidative stress in mice with NAFLD, which plays a critical role in the occurrence and development of NAFLD. In addition, PYOs could markedly decrease lipid accumulation in liver by activating the IRS-1/AKT/GSK-3β signaling pathway and the AMPK signaling pathway in mice with NAFLD. PYOs also apparently relieved the hepatic fibrosis induced by oxidative stress via downregulation of TGF-β and its related proteins, so that liver injury was markedly alleviated. Furthermore, PYOs treatment relieved cecal microbiota dysbiosis (such as increasing the relative abundance of Akkermansia, while decreasing the Helicobacter abundance), which could alleviate oxidative stress, inflammation, and lipid metabolism, and protect the liver to a certain degree. In summary, PYOs treatment remarkably improved NAFLD via a specific molecular mechanism and reshaped the cecal microbiota.

Topics: Animals; Cecum; Disease Models, Animal; Dysbiosis; Gastrointestinal Microbiome; Lipid Metabolism; Male; Mice; Mice, Inbred C57BL; Non-alcoholic Fatty Liver Disease; Oligosaccharides; Oxidative Stress; Sepharose; Signal Transduction

Topics: Aerobiosis; Anti-Infective Agents; Antioxidants; Benzothiazoles; Biphenyl Compounds; Escherichia coli; Microbial Sensitivity Tests; Oxidation-Reduction; Picrates; Porphyra; Proton Magnetic Resonance Spectroscopy; Sepharose; Spectrophotometry, Ultraviolet; Spectroscopy, Fourier Transform Infrared; Staphylococcus aureus; Sulfonic Acids; Tyrosine

An antioxidant polysaccharide, porphyran, from red algae Pyropia haitanensis, is introduced as a protective agent against neurotoxicity-induced amyloid β peptide (Aβ) of Alzheimer's disease (AD) mice. Then the activity of acetylcholinesterase (AChE) and choline acetyltransferase (CHAT) in the cortical and hippocampal tissue were examined by colorimetric method. Results showed that porphyran significantly ameliorated the learning and memory impairment induced by Aβ

Topics: Acetylcholine; Acetylcholinesterase; Alzheimer Disease; Amyloid beta-Peptides; Animals; Antioxidants; Choline O-Acetyltransferase; Hippocampus; Male; Maze Learning; Memory; Memory Disorders; Mice; Neuroprotective Agents; Peptide Fragments; Polysaccharides; Rhodophyta; Sepharose

We previously demonstrated that porphyran, a sulfated polysaccharide extracted from Pyropia yezoensis, shows protective effects on LPS-induced septic shock in the mouse. However, the immune cell-mediated inhibitory effect of porphyran in LPS-induced activation of immune cells has not been well investigated. In this study, we found that treatment of porphyran suppressed LPS-induced upregulation of costimulatory molecule and C-C chemokine receptor type 7 (CCR7) expression in bone marrow-derived dendritic cells (BMDCs) in vitro and spleen DCs in vivo. Moreover, the LPS-induced expression of IL-6, IL-12, and TNF-α in the culture medium of BMDCs and serum dose-dependently decreased by porphyran treatment, which contributed to the inhibition of the intracellular cytokine production in spleen DCs. In addition, LPS-induced differentiation of helper T1 (Th1) and cytotoxic T1 (Tc1) cells was effectively suppressed by porphyran treatment in mice. The inhibitory effect of porphyran in LPS-induced immune activation was mediated by competitive binding of porphyran with LPS in spleen DCs. Thus, these results suggest that porphyran is a promising potential therapeutic agent in endotoxin-mediated inflammatory disease and septic shock.

Topics: Animals; Anti-Inflammatory Agents; Bone Marrow Cells; Cell Differentiation; Cytokines; Dendritic Cells; Immunity, Cellular; Lipopolysaccharides; Mice, Inbred C57BL; Polysaccharides, Bacterial; Rhodophyta; Sepharose; Spleen; T-Lymphocytes, Cytotoxic; Th1 Cells

In mice, porphyran extracted from Pyropia yezoensis exerts anti-inflammatory effects and suppresses lipopolysaccharide (LPS)-induced immune activation and sepsis. Here, we investigated inhibition of LPS-induced activation of human immune cells by porphyran and the underlying mechanisms. We demonstrated that porphyran may inhibit LPS-induced proinflammatory cytokine production in peripheral blood mononuclear cells, monocyte-derived dendritic cells, and peripheral blood dendritic cells. We also observed that porphyran-mediated LPS-induced activation of DC suppressed syngeneic T cell proliferation, resulting in reduced production of interferon-γ. The mechanism of LPS-induced immune cell activation requires the activation of toll like receptor 4 following binding of LPS to myeloid differentiation factor 2 (MD2). Additionally, we show that porphyran competes with LPS for binding to MD2 and thereby suppresses immune cell activation. Porphyran may, therefore, be a promising therapeutic candidate for the treatment of endotoxin-mediated septic shock and inflammation in humans.

Topics: Cytokines; Dendritic Cells; Humans; Leukocytes, Mononuclear; Lipopolysaccharides; Sepharose

The oligosaccharides from agar hydrolysis have special biological activities, and exhibit application prospects in cosmetic, food and pharmaceutical industry. In this study, two novel β-agarases (AgaA and AgaB) were screened and characterized. It was found that the AgaA was an endo-type agarase which could efficiently hydrolyzed agar or agarose to form neoagarobiose (NA2), neoagarotetraose (NA4) and neoagarohexaose (NA6), while the AgaB was an exo-type and bifunctional enzyme that showed activities towards both agarose and porphyran. Based on the properties of the two enzymes, we developed modular strategy for enzymatic production of neoagarobiose through a two-stage hydrolysis reaction. The cheap substrate agar was first liquefied by AgaA at high temperature to form neoagaroligosaccharides, which together with the sulfated polysaccharides were homogenized by AgaB to form neoagarobiose as the final product. High concentration of agar (10 g/L) was almost completely converted into neoagarobiose with high purity.

Topics: Agar; Disaccharides; Galactosides; Glycoside Hydrolases; Hydrolysis; Oligosaccharides; Sepharose

Porphyran is the major polysaccharide of laver and mainly composed of 3-linked β-d-galactopyranose (G) and 4-linked α-l-galactopyranose-6-sulfate (L6S) units. Structural heterogeneity of porphyran highly originates from the natural methylation on the O-6 position of G units (GMe). Here, a GH16 porphyranase Por16C_Wf was cloned from a porphyran-related polysaccharide utilization locus of

Topics: Bacterial Proteins; Flavobacteriaceae; Galactose; Glycoside Hydrolases; Multigene Family; Sepharose; Substrate Specificity

Topics: Animals; cdc42 GTP-Binding Protein; Cell Line; Cell Movement; Cell Proliferation; Cell Survival; Epithelial Cells; Focal Adhesion Kinase 1; Gastrointestinal Agents; Gene Expression Regulation; Intestinal Mucosa; Molecular Weight; Paxillin; Porphyra; Pseudopodia; Rats; Sepharose; Viscosity

In the present work, the green synthesis of silver nanoparticles (AgNPs) using the sulfated polysaccharide porphyran (PFR) as capping agent and d-glucose as reducing agent is described. PFR was extracted from red seaweed and characterized by employing

Topics: Anti-Bacterial Agents; Electrochemistry; Electrodes; Fluorouracil; Metal Nanoparticles; Sepharose; Silver

Pyropia yezoensis, rich in porphyran, is a medicine-edible red alga. In the present study, the physicochemical characteristics, conformational states and antitumor activities of a novel porphyran extracted from the high-yield algal strain Pyropia yezoensis Chonsoo2 and its two degraded derivatives by gamma irradiation were investigated.. Pyropia yezoensis porphyran is a water-soluble, triple-helical sulfated hetero-galactopyranose, named PYP. PYP was degraded by gamma irradiation at 20 kGy and 50 kGy, giving two low molecular weight derivatives comprising PYP-20 and PYP-50, respectively. PYP with a higher molecular weight has a solution conformation different from PYP-20 and PYP-50. Three porphyrans had no toxicity in normal human liver cells (HL-7702) and showed antitumor effects on Hep3B, HeLa and MDA-MB-231. They had better antitumor against HeLa cells, exhibiting a similar inhibition ratio compared to 5-fluorouracil, with PYP especially exhibiting a higher inhibition ratio than 5-fluorouracil. With respect to HeLa cells, the different antitumor activities might be related to porphyran molecular weight and solution conformation. Furthermore, the HeLa cell cycle was blocked in the G2/M phase after PYP treatment, leading to cell proliferation inhibition. The induction of cell cycle arrest was related to the changes in the expression of p21, p53, Cyclin B1 and cyclin-dependent kinase 1.. Pyropia yezoensis porphyran, as applied to medicine and functional food, could potentially be used as a non-toxic natural adjuvant in cancer therapy. © 2019 Society of Chemical Industry.

Topics: Antineoplastic Agents; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cyclin B1; Gene Expression Regulation, Neoplastic; Humans; Plant Extracts; Rhodophyta; Sepharose; Tumor Suppressor Protein p53

Topics: Amino Acid Sequence; Bacterial Proteins; Biocatalysis; Biotechnology; Cloning, Molecular; Enzyme Stability; Flavobacteriaceae; Glycoside Hydrolases; Hydrogen-Ion Concentration; Hydrolysis; Molecular Weight; Phylogeny; Porphyra; Seawater; Sepharose; Sequence Alignment

Safe and efficient therapeutic agents for bone diseases are required in natural sources. We previously found that edible seaweed-derived polysaccharide porphyran exhibited anti-inflammatory effects through the down regulation of nuclear factor-κB. The aim of this study was to investigate the availability of porphyran as a therapeutic agent for bone diseases. The effects of porphyran on receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclastogenesis in RAW264.7 cells were examined. Porphyran suppressed RANKL-induced osteoclast formation in a concentration-dependent manner (6.25-50 μg/ml) without any cytotoxic effects. Furthermore, real-time polymerase chain reaction analyses indicated that porphyran at 50 μg/ml significantly attenuated the RANKL-induced increase in the mRNA levels of osteoclastogenesis-related marker genes such as nuclear factor of activated T cells, tartrate-resistant acid phosphatase, cathepsin K, and matrix metalloproteinase-9 in RAW264.7 cells. To our knowledge, this is the first report showing that edible-seaweed-derived polysaccharide porphyran can suppress RANKL-induced osteoclastogenesis. Our results suggest that porphyran can be used as a safe therapeutic agent to improve osteoclast-related pathological conditions.

Topics: Animals; Cell Differentiation; Mice; Osteoclasts; RANK Ligand; RAW 264.7 Cells; Sepharose

Sugar O-methylation shields algal polysaccharides against microbial hydrolytic enzymes. Here, we describe cytochrome P450 monooxygenases from marine bacteria that, together with appropriate redox-partner proteins, catalyze the oxidative demethylation of 6-O-methyl-D-galactose, which is an abundant monosaccharide of the algal polysaccharides agarose and porphyran. This previously unknown biological function extends the group of carbohydrate-active enzymes to include the class of cytochrome P450 monooxygenases.

Topics: Bacteria; Carbohydrates; Cloning, Molecular; Computational Biology; Cytochrome P-450 Enzyme System; Demethylation; Gas Chromatography-Mass Spectrometry; Hexoses; Hydrogen Peroxide; Methylation; NAD; Oxidation-Reduction; Phylogeny; Polysaccharides; Rhodophyta; Sepharose; Substrate Specificity

Topics: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; Animals; Apoptosis; Behavior, Animal; Brain; Disease Models, Animal; Dopamine; Dopaminergic Neurons; Humans; Male; Mice; Mice, Inbred C57BL; MPTP Poisoning; Neuroprotective Agents; Phosphatidylinositol 3-Kinases; Porphyra; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Sepharose; Signal Transduction

The dense microbial ecosystem in the gut is intimately connected to numerous facets of human biology, and manipulation of the gut microbiota has broad implications for human health. In the absence of profound perturbation, the bacterial strains that reside within an individual are mostly stable over time

Topics: Animals; Bacteroides; Colon; Fecal Microbiota Transplantation; Female; Gastrointestinal Microbiome; Humans; Male; Mice; Sepharose

The anti-inflammatory properties of porphyrans (D1-D4) obtained from four discolored nori (Pyropia yezoensis) with different growth backgrounds were studied to examine possible variations in their bioactivities. Elution profiles of the porphyrans on Sepharose 4B indicated that D2-porphyran had relatively lower-molecular-size porphyrans than the other porphyrans. Inhibitory activities of the four porphyrans against nitric oxide (NO) and tumor necrosis factor-α (TNF-α) secretion by lipopolysaccharide (LPS)-stimulated RAW264.7 cells were different, whereas no significant differences were observed in the sulfate and anhydrogalactose levels. D2-porphyran showed the highest inhibitory activity against NO and TNF-α secretion by LPS-stimulated RAW264.7 cells, whereas D3- and D4-porphyrans had almost no activity. All porphyrans were efficiently degraded by free radical generated with ascorbate and hydrogen peroxide. The free-radical degradation resulted in a significant increase in the inhibitory activities of the four porphyrans against NO and TNF-α secretion, with varying rates depending on the porphyrans. The ability of D2-porphyran to suppress the receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclastogenesis in RAW264.7 cells was also significantly enhanced after degradation. Our results suggest that molecular size is an important factor affecting the anti-inflammatory activity of porphyrans, and radical degradation might be a promising procedure to obtain active low-molecular-size porphyrans.

Topics: Animals; Anti-Inflammatory Agents; Cell Survival; Chromatography, Gel; Macrophages; Mice; Nitric Oxide; Phytochemicals; Plant Extracts; Porphyra; RANK Ligand; RAW 264.7 Cells; Sepharose

Porphyran is a sulfate galactan in the cell wall of

Topics: Acute Kidney Injury; Animals; Blood Urea Nitrogen; Creatinine; Female; Glycerol; Ions; Kidney; Male; Oligosaccharides; Polymerization; Protective Agents; Rats; Rats, Sprague-Dawley; Sepharose

The hypolipidemic and antioxidant effects of porphyran from the red algae Porphyra haitanensis as a dietary supplement were evaluated in mice. The levels of serum TC, TG and LDL-C in MP group increased significantly (p<0.05 or p<0.01) by 28.5%, 29.4% and 33.5% compared with the model group. These significant rises were accompanied by significant declines of plasma HDL-C by 21.6% compared with the model group. In addition, the liver content of malondialdehyde significantly decreased, while the superoxide dismutase, catalase, glutathione peroxidase activities significantly increased. The levels of serum SOD and GSH-Px in MP group increased significantly by 51.2% and 99.6% compared with the model group. The results suggested that porphyran could be used as functional foods and natural drugs in preventing the hyperlipidemia and this activity might be attributed to its antioxidant potential.

Topics: Animals; Antioxidants; Catalase; Glutathione Peroxidase; Hyperlipidemias; Hypolipidemic Agents; Male; Malondialdehyde; Mice; Porphyra; Sepharose; Superoxide Dismutase

The activation of ions by extreme-energy photons (XUV) produced by a synchrotron radiation beamline is a powerful method for characterizing complex glycans using tandem mass spectrometry (MS). As previously described, this activation method leads to rich fragmentation spectra with many structurally valuable cross-ring cleavages while maintaining labile modifications on the glycan structures. However, until now, the tandem MS event was too long to be compatible with liquid chromatography elution times. In this work, the duty cycle of the activation and detection of fragments was shortened, and the background signal on the spectra was drastically reduced. Both improvements allowed, for the first time, the successful coupling of a UHPLC system to XUV-activated tandem MS. The approach was used to characterize a complex mixture of oligo-porphyrans, which are a class of highly sulfated oligosaccharides, in a fully automated way. Due to an enhanced dynamic range and an increased sensitivity, some hypothetical structures of low abundance have been unequivocally confirmed in this study and others have been revised. Some previously undescribed species of oligo-porphyrans that exhibit lateral branching have been fully resolved. This work contributes to the scarce knowledge of the structure of porphyrans in red algae and pushes the current capacities of XUV-activation tandem MS by demonstrating the possibility of a direct coupling with UHPLC. This study will considerably broaden the applicability and practicality of this method in many fields of analytical biology.

Topics: Chromatography, High Pressure Liquid; Photons; Polysaccharides; Sepharose; Tandem Mass Spectrometry; Ultraviolet Rays

Porphyran, a sulfated polysaccharide, isolated from discolored nori (Porphyra yezoensis) (dc-porphyran) and one fraction (F1) purified from dc-porphyran by DEAE-chromatography showed the protective effects on LPS-induced endotoxin shock in mice. Intraperitoneal (i.p.) treatment with dc-porphyran or F1 (100mg/kg) 60min prior to i.p. injection of LPS (30mg/kg) completely protected mice from LPS lethality. At 10mg/kg concentration, F1 demonstrated more protection than dc-porphyran. Intravenous (i.v.) challenge of LPS, even at 20mg/kg, was more lethal than i.p. administration; i.v. injection of F1 (100mg/kg) with LPS significantly improved the survival rate. However, i.v. dc-porphyran (100mg/kg) produced an even lower survival rate than that of LPS alone. We examined pro-inflammatory mediators such as NO and TNF-α in serum. F1 significantly reduced the levels of these markers. Additionally, F1 significantly decreased the malondialdehyde level in the liver, a marker of oxidative stress, while dc-porphyran had almost no effect. Furthermore, F1 significantly decreased the production of TNF-α and NO in peritoneal exudate cells harvested from LPS-challenged mice, while dc-porphyran treatment showed a lesser decrease. Our results suggest that porphyran isolated from discolored nori, especially F1, is capable of suppressing LPS-induced endotoxin shock in vivo.

Topics: Animals; Color; Lipopolysaccharides; Liver; Male; Mice; Nitric Oxide; Oxidative Stress; Porphyra; Sepharose; Shock, Septic; Tumor Necrosis Factor-alpha

Extreme ultraviolet photon activation tandem mass spectrometry (MS) at 69 nm (18 eV) was used to characterize mixtures of oligo-porphyrans, a class of highly sulfated oligosaccharides. Porphyrans, hybrid polymers whose structures are far from known, continue to provide a challenge for analytical method development. Activation by 18 eV photons led to a rich fragmentation of the oligo-porphyrans, with many cross-ring and glycosidic cleavages. In contrast to multistage MSn strategies such as activated electron photodetachment dissociation, a single step of irradiation by energetic UV of multiply charged anions led to a complete fragmentation of the oligo-porphyrans. In both ionization modes, the sulfate groups were retained on the backbone, which allowed the pattern of these modifications along the porphyran backbone to be described in unprecedented detail. Many structures released by the enzymatic degradation of the porphyran were completely resolved, including isomers. This work extends the existing knowledge of the structure of porphyrans. In addition, it provides a new demonstration of the potential of activation by high-energy photons for the structural analysis of oligosaccharides, even in unseparated mixtures, with a particular focus on sulfated compounds.

Topics: Cell Wall; Chromatography, High Pressure Liquid; Ions; Oligosaccharides; Photons; Porphyra; Sepharose; Sulfates; Tandem Mass Spectrometry

We found that discolored waste nori with no commercial value, contains much higher level of porphyran than normal nori that is a sheeted food stuff prepared from P. yezoensis used in sushi. Chemical analyses revealed that mean molecular mass of the porphyran prepared from discolored nori (dc-porphyran) was much lower than that of the porphyran from normal nori (n-porphyran). Dc-porphyran showed slightly greater scavenging activity toward superoxide anion and hydroxyl radical than n-porphyran. Dc-porphyran inhibited nitric oxide (NO) production in LPS-stimulated RAW264.7 cells through preventing the expression of inducible NO synthase, whereas no such activity was observed in n-porphyran. Since acid-hydrolyzed n-porphyran showed the inhibitory activity on NO production from LPS-stimulated RAW264.7 cells, the molecular size of porphyran was suggested to be a critical factor for the activity. Dc-porphyran was separated into 4 fractions (F1-F4) on DEAE-chromatography, and F1 showed the highest inhibitory effect on NO production from LPS-stimulated RAW264.7 cells. Our results indicate that discolored waste nori is useful as a source of porphyran with even better bioactivities than porphyran from normal nori.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Cell Line; Free Radical Scavengers; Macrophages; Mice; Nitric Oxide; Nitric Oxide Synthase Type II; Plant Extracts; Porphyra; Reactive Oxygen Species; Sepharose; Tumor Necrosis Factor-alpha

The influence of sulfated polysaccharides (λ-, κ-, and κ/β-carrageenan and porphyran) - on platelet activation was studied. Carrageenans were much weaker inhibitors of a coagulation process than heparin, while porphyran had not that effect. Results of the aPTT and PT assays suppose that carrageenans affected mostly intrinsic pathway of coagulation, while their effect on the extrinsic pathway is extremely low (λ and κ/β) or absent (κ, LMW derivative of κ-carrageenan). λ-Carrageenan was the most potent anticoagulant agent in TT, aPTT, PT, and anti-factor Xa activity. This sample was also the strongest inhibitor of collagen-induced platelet aggregation in PRP. Generally, the correlation of anticoagulant and antithrombotic action in PRP is preserved for carrageenans but not for heparin. Carrageenans and porphyran affected platelet adhesion to collagen by influencing glycoprotein VI. Low molecular weight κ-carrageenan had a similar effect on platelet adhesion mediated with both major collagen receptors: integrin α2 β1 and glycoprotein VI as native polysaccharide had. Carrageenans resulted in activation of platelets under platelet adhesion mediated by integrin αIIb β3 with less degree than heparin. The least sulfated κ/β-carrageenan that possessed an inhibiting effect on thrombin- and collagen-induced aggregation of washed platelets and on the PT test but it had no significant effect on TT was the weakest promoter of integrin αIIb β3 mediated platelet activation. In summary, our study showed that the polysaccharide action was complex, since it depended on its molecular mass, sulfation degree, and monosaccharide contents (3,6-anhydrogalactose).

Topics: Animals; Blood Coagulation; Blood Platelets; Carrageenan; Cattle; Collagen; Erythrocytes; Factor Xa; Fibrinogen; Hemagglutination Tests; Hemolysis; Humans; Partial Thromboplastin Time; Platelet Activation; Platelet Adhesiveness; Platelet Aggregation; Polysaccharides; Prothrombin Time; Rhodophyta; Sepharose; Thrombin Time

Development of oral amphotericin B (AmB) loaded nanoparticles (NPs) demands a novel technique which reduces its toxicity and other associated problems. Packing of AmB in between two oppositely charged ions by polyelectrolyte complexation technique proved to be a successful strategy. We have developed a novel carrier system in form of polyelectrolyte complex of AmB by using chitosan (CS) and porphyran (POR) as two oppositely charged polymers with TPP as a crosslinking agent. Initially POR was isolated from Porphyra vietnamensis followed by the fact that its alkali induced safe reduction in molecular weight was achieved. Formulation was optimized using three-factor three-level (3(3)) central composite design. High concentration of POR in NPs was confirmed by sulfated polysaccharide (SP) assay. Degradation and dissolution studies suggested the stability of NPs over wide pH range. Hemolytic toxicity data suggested the safety of prepared formulation. In vivo and in vitro antifungal activity demonstrated the high antifungal potential of optimized formulation when compared with standard drug and marketed formulations. Throughout the study TPP addition did not cause any significant changes. Therefore, these experimental oral NPs may represent an interesting carrier system for the delivery of AmB.

Topics: Administration, Oral; Amphotericin B; Antifungal Agents; Chitosan; Drug Carriers; Humans; Hydrogen-Ion Concentration; Nanoparticles; Polymers; Sepharose

Our investigation explores the immuno-efficiency of sulphated polysaccharides enriched Porphyra vietnamenis. Isolated polysaccharide fraction (17.1-25.8%) was characterized by FTIR and NMR spectroscopy which showed the presence of typical linear backbone structure called as porphyran. Oral administration of porphyran (200-500 mg/kg) evoked a significant (P ≤ 0.05) increase in weight of the thymus, spleen and lymphoid organ cellularity. The total leucocyte and lymphocyte count was increased significantly (P<0.005). The increase in the percent neutrophil adhesion to nylon fibres as well as a dose-dependent increase in antibody titre values was observed. A decreased response to DTH reaction induced by SRBC was recorded. A potential phagocytic response was seen and significant changes were observed in the formation of formazone crystals. It also prevented myelosuppression in cyclophosphamide drug treated rats. The results indicated that P. vietnamenis possesses potential immunomodulatory activity and has therapeutic potential for the prevention of autoimmune diseases.

Topics: Administration, Oral; Animals; Antibody Formation; Autoimmune Diseases; Cell Adhesion; Immunologic Factors; Lymphocyte Count; Mice; Neutrophils; Porphyra; Rats; Rats, Wistar; Sepharose

Porphyran, extracted from an edible red alga (Porphyra yezoensis), is a sulphated polysaccharide with a wide variety of biological activities including anti-tumour, antioxidant and immuno-modulating activities. In this study, we examined the effect of porphyran on nitric oxide (NO) production in mouse macrophage cell line RAW264.7 cells. Although no significant activity of porphyran to induce NO or tumour necrosis factor-α (TNF-α) production in RAW264.7 cells was observed at the concentration range tested (10-500 µg/ml), it was found for the first time that porphyran inhibited NO production and expression of inducible nitric oxide synthase (iNOS) in RAW264.7 cells stimulated with lipopolysaccharide (LPS). In the presence of 500 µg/ml porphyran, NO production and expression of iNOS in LPS-treated RAW264.7 cells were completely suppressed. On the other hand, porphyran showed only a marginal effect on the secretion of TNF-α from LPS-stimulated RAW264.7 cells. Electrophoretic mobility shift assay (EMSA) using infrared dye labelled oligonucleotide with nuclear factor-κB (NF-κB) consensus sequence suggested that porphyran inhibited the LPS-induced NF-κB activation. The LPS-inducible nuclear translocation of p65, and the phosphorylation and degradation of IκB-α were also inhibited by the pre-treatment with porphyran. Our results obtained in in vitro analysis suggest that porphyran suppresses NO production in LPS-stimulated macrophages by the blocking of NF-κB activation.

Topics: Active Transport, Cell Nucleus; Animals; Cell Line; Cell Nucleus; Cell Survival; DNA Fragmentation; Dose-Response Relationship, Drug; Gene Expression Regulation, Enzymologic; I-kappa B Proteins; Lipopolysaccharides; Macrophages; Mice; NF-kappa B; NF-KappaB Inhibitor alpha; Nitric Oxide; Nitric Oxide Synthase Type II; Phosphorylation; Reverse Transcriptase Polymerase Chain Reaction; Sepharose; Superoxides; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Porphyran (POR) from the red alga Porphyra yezoensis is a water soluble dietary fiber. In this study, we investigated the effect of dietary POR on glucose metabolism in KK-Ay mice (a model for type 2 diabetes). Mice were divided into 4 groups and fed a diet containing 5% cellulose (control), POR, POR Arg or POR K. After 3 wk of feeding, plasma insulin levels and the calculated homeostasis model assessment-insulin resistance (HOMA-IR) index were significantly lower in the POR group than in the control group. Compared with the control group, plasma adiponectin levels were significantly increased in the POR, POR Arg and POR K groups. These results suggest that dietary POR should improve glucose metabolism in diabetes via up-regulation of adiponectin levels. In addition, the amount of propionic acid in the cecum of the POR group was significantly higher than in the control group and the profile of bacterial flora was changed by dietary POR. In the cecum of the POR, POR Arg and POR K groups, Bacteroides was significantly increased and Clostridium coccoides was significantly decreased compared with in the control group. The effects of dietary POR on the hindgut environment might contribute to the improvement of glucose metabolism.

Topics: Adiponectin; Animals; Cecum; Cholesterol; Diabetes Mellitus, Type 2; Dietary Fiber; Disease Models, Animal; Fatty Acids, Nonesterified; Glucose; Insulin; Insulin Resistance; Male; Mice; Porphyra; Real-Time Polymerase Chain Reaction; RNA, Bacterial; Sepharose; Triglycerides

In the present study, we have explored porphyran as a reducing agent for one pot size controlled green synthesis of gold nanoparticles (AuNps) and further investigated its application as a carrier for the delivery of an anticancer drug. The prepared AuNps showed surface plasmon resonance centered at 520 nm with average particle size of 13±5 nm. FTIR spectra suggested that the sulfate moiety is mainly responsible for reduction of chloroauric acid. The capping of the AuNps with porphyran was evident from the negative zeta potential value responsible for the electrostatic stability. Thus, porphyran acts as reducing as well as capping agent. These AuNps are highly stable in a wide range of pH and electrolyte concentration. Porphyran capped AuNps exhibited enhanced cytotoxicity on human glioma cell line (LN-229) as compared to native porphyran. Consequently, these AuNps have been utilized as a carrier for delivery of the anticancer drug doxorubicin hydrochloride (DOX). Spectroscopic examination revealed that DOX conjugated onto AuNps via hydrogen bonding. The release of DOX from DOX loaded AuNps was found to be sixfold higher in acetate buffer (pH 4.5) as compared to physiological buffer (pH 7.4). Further, the DOX loaded AuNps demonstrated higher cytotoxicity on LN-229 cell line as compared with an equal dose of native DOX solution. This established the potential of these AuNps as a carrier for anticancer drug delivery.

Topics: Antibiotics, Antineoplastic; Cell Line, Tumor; Chlorides; Doxorubicin; Drug Carriers; Drug Delivery Systems; Glioma; Gold; Gold Compounds; Humans; Hydrogen Bonding; Hydrogen-Ion Concentration; Metal Nanoparticles; Particle Size; Sepharose; Spectroscopy, Fourier Transform Infrared; Static Electricity; Surface Plasmon Resonance

Topics: Adaptation, Physiological; Bacteroides; Biological Evolution; Cultural Diversity; Diet; Feces; Food Microbiology; Gene Transfer, Horizontal; Glycoside Hydrolases; Humans; Intestines; Japan; Marine Biology; Metagenome; Porphyra; Sepharose; United States

Gut microbes supply the human body with energy from dietary polysaccharides through carbohydrate active enzymes, or CAZymes, which are absent in the human genome. These enzymes target polysaccharides from terrestrial plants that dominated diet throughout human evolution. The array of CAZymes in gut microbes is highly diverse, exemplified by the human gut symbiont Bacteroides thetaiotaomicron, which contains 261 glycoside hydrolases and polysaccharide lyases, as well as 208 homologues of susC and susD-genes coding for two outer membrane proteins involved in starch utilization. A fundamental question that, to our knowledge, has yet to be addressed is how this diversity evolved by acquiring new genes from microbes living outside the gut. Here we characterize the first porphyranases from a member of the marine Bacteroidetes, Zobellia galactanivorans, active on the sulphated polysaccharide porphyran from marine red algae of the genus Porphyra. Furthermore, we show that genes coding for these porphyranases, agarases and associated proteins have been transferred to the gut bacterium Bacteroides plebeius isolated from Japanese individuals. Our comparative gut metagenome analyses show that porphyranases and agarases are frequent in the Japanese population and that they are absent in metagenome data from North American individuals. Seaweeds make an important contribution to the daily diet in Japan (14.2 g per person per day), and Porphyra spp. (nori) is the most important nutritional seaweed, traditionally used to prepare sushi. This indicates that seaweeds with associated marine bacteria may have been the route by which these novel CAZymes were acquired in human gut bacteria, and that contact with non-sterile food may be a general factor in CAZyme diversity in human gut microbes.

Topics: Adaptation, Physiological; Bacteroides; Biological Evolution; Crystallography, X-Ray; Cultural Diversity; Diet; Eukaryota; Feces; Food Microbiology; Gene Transfer, Horizontal; Genome, Bacterial; Glycoside Hydrolases; Humans; Intestines; Japan; Marine Biology; Metagenome; Models, Molecular; North America; Phylogeny; Porphyra; Protein Conformation; Sepharose; Substrate Specificity

The physiological effect of porphyran, a sulfated polysaccharides from an edible red alga, was studied in human hepatoma HepG2 cells. Porphyran supplementation significantly decreased apolipoprotein B100 secretion, and the reduction was partly associated with suppression of cellular lipid synthesis in HepG2 cells. This is the first study to elucidate the mechanism of the hypolipidemic effect of porphyran.

Topics: Animals; Apolipoprotein B-100; Cattle; Cell Line, Tumor; Cholesterol Esters; Humans; Lipids; Seaweed; Sepharose; Sulfur; Triglycerides

Porphyran extracted from Porphyra haitanensis is a sulfated polysaccharide, which possesses excellent antioxidant activities. In this study, we prepared one low-molecular-weight porphyran and its sulfated, acetylated, phosphorylated and benzoylated derivatives. Their antioxidant activities were investigated including scavenging effect of superoxide, hydroxyl and 1,1-diphenyl-2-picrylhydrazyl radicals. The results of chemical analysis and FT-IR spectrums showed the modification was successful. And in addition, we found that certain derivative exhibited stronger antioxidant activity than low-molecular-weight porphyran. The benzoylated derivative showed the most excellent antioxidant activity in three assays, so this derivative needs to be attended to.

Topics: Antioxidants; Biphenyl Compounds; China; Hydroxyl Radical; Picrates; Porphyra; Sepharose; Spectroscopy, Fourier Transform Infrared; Superoxides

Porphyran (Por) prepared from dried nori was applied as a functional modifier of a soybean protein isolate (SPI) to conjugate with SPI from defatted soybean by the Maillard reaction (79% relative humidity and 60 degrees C for 7 days). Two kinds of partially denatured conjugate (Conj 45 and Conj 63) were obtained from the reaction product by sequential extraction at pH 4.5 and pH 6.3, and the respective yield and weight ratios of the SPI and Por moieties were 8.4% and 1:1 for Conj 45 and 11.7% and 1:0.16 for Conj 63. Conj 63 demonstrated improved solubility between pH 5.0 and pH 8.0, while Conj 45 exhibited substantially complete solubility over the pH range of 2.0-8.0. Conj 63 showed more tolerance against digestion with pancreatin than SPI, whereas this was lost after denaturation. Conj 63 and Conj 45 both showed a markedly higher emulsion activity index and emulsion stability than SPI, even at pH 3.0; in particular, Conj 45 exhibited outstanding emulsifying ability. Conj 63 had about a two-fold higher calcium-binding ability than SPI, and Conj 63 and Conj 45 did not aggregate with added Ca2+ and Mg2+. It is believed that Por could be a valuable functional modifier of SPI for providing soybean protein-based liquid foods such as beverages by conjugation through the Maillard reaction.

Topics: Chemical Phenomena; Chemistry, Physical; Emulsifying Agents; Maillard Reaction; Pancreatin; Protein Denaturation; Rhodophyta; Sepharose; Solubility; Soybean Proteins

In the present paper, ascorbate and hydrogen peroxide (H2O2) were used to degrade porphyran. It was found that porphyran could be degraded by free radical that was generated by ascorbate and H2O2 in combination. It was possible to prepare desired porphyran products with different molecular weight by adjusting ascorbate to H2O2 proportions and their concentrations. The molar ratio of 1 was demonstrated more effective than in other ratios. Higher concentrations accelerated the degradation. Moreover, results of chemical analysis and FT-IR spectra suggested that the main structure of degraded products still remained although some changes happened. The degraded and natural porphyrans possessed scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH)-radical activity and reducing power. Higher antioxidant activities were found in both systems when the molecular weight was reduced. The results indicated that the antioxidant activities were closely related to the molecular weight. The degraded porphyrans are potential antioxidant in vitro.

Topics: Antioxidants; Ascorbic Acid; Biodegradation, Environmental; Free Radicals; Hydrogen Peroxide; Molecular Weight; Porphyra; Sepharose

Marine microorganisms degrading porphyran (POR) were found on the surface of thalli of Porphyra yezoensis. Fifteen crude microorganism groups softened and liquefied the surface of agar-rich plate medium. Among these, 11 microorganism groups degraded porphyran that consisted of sulfated polysaccharide in Porphyra yezoensis. Following isolation, 7 POR-degradable microorganisms were isolated from the 11 POR-degradable microorganism groups.

Topics: Molecular Weight; Oceans and Seas; Porphyra; Sepharose

Porphyrans, the sulfated polysaccharides, are the main components of Porphyra. The potential apoptotic activities of porphyran were evaluated using AGS human gastric cancer cells. Porphyran did not affect the growth of normal cells, but did induce cancer cell death in a dose-dependent manner. The addition of 0.1% porphyran also reduced DNA synthesis after 24 h of exposure, suggesting that porphyran inhibits cancer cell growth by both decreasing cell proliferation and inducing apoptosis. AGS cells treated with porphyran displayed a marked increase in poly(ADP-ribose) polymerase (PARP) cleavage, as well as caspase-3 activation. The ability of porphyran to promote apoptosis may contribute to its usefulness as an agent capable of significantly inhibiting cell growth in AGS human gastric cancer cells. Insulin-like growth factor-I receptor (IGF-IR) phosphorylation was decreased in porphyran-treated AGS cells compared to control cells, which correlated with Akt activation. Thus, porphyran appears to negatively regulate IGF-IR phosphorylation by causing a decrease in the expression levels in AGS gastric cancer cells, and then inducing caspase-3 activation.

Topics: Apoptosis; Caspase 3; Caspases; Cell Line, Tumor; Cell Proliferation; DNA Replication; Enzyme Activation; Humans; Phosphorylation; Poly(ADP-ribose) Polymerases; Proto-Oncogene Proteins c-akt; Receptor, IGF Type 1; Sepharose; Signal Transduction; Stomach Neoplasms

The nucleotide sequence of the gene for the alpha-agarase, AgaA33, from Thalassomonas sp. strain JAMB-A33 was determined. The open reading frame for AgaA33 was revealed to encode 1463 amino acid residues. We succeeded in extracellular production of recombinant -agarase (AgaA33) efficiently using Bacillus subtilis as a host. This is the first report of recombinant production of -agarase. Furthermore, we demonstrated that hydrolysis of alpha-1,3 linkages in porphyran, a sulfated polysaccharide from marine red algae, by alpha-agarase is an important step for improvement of its antioxidant activity with regard to free-radical-scavenging capacity and superoxide radical anion scavenging activity, whereas the hydrolysis of beta-1,4 linkages in porphyran by beta-agarase did not increase on the antioxidant activity markedly.

Topics: Amino Acid Sequence; Antioxidants; Base Sequence; Carbohydrate Conformation; DNA, Bacterial; Gammaproteobacteria; Glycoside Hydrolases; Hydrolysis; Molecular Sequence Data; Open Reading Frames; Recombinant Proteins; Rhodophyta; Sepharose

Porphyran is a major component of the red algae, Porphyra tenera and P. yezoensis, which are processed into a sheet type of dried food, "Nori". Porphyran has been reported to activate murine macrophages by in vitro and i.p. injection studies. The contact hypersensitivity (CHS) reaction in mice is commonly used as a model to evaluate the anti-allergic activity of food and food components. We therefore studied the effect of porphyran on the CHS reaction in Balb/c mice to evaluate anti-allergic activity of porphyran. We found that an oral administration of porphyran (2% in drinking water) suppressed the CHS reaction (ear edema) induced by 2,4,6-trinitrochlorobenzene. We also found that porphyran suppressed the serum level of IgE and the production of interferon-gamma (IFN-gamma) in the challenged ear lobe. We conclude from these results that the CHS reaction was suppressed by oral porphyran due to the decreased serum level of IgE and the production of IFN-gamma in the challenged ear lobe.

Topics: Animals; Anti-Allergic Agents; Dermatitis, Contact; Ear; Edema; Immunoglobulin E; Immunosuppression Therapy; Interferon-gamma; Mice; Mice, Inbred BALB C; Picryl Chloride; Porphyra; Sepharose

Three porphyran preparations with high emulsifying ability and varying molecular mass, 3,6-anhydrogalactose content, and sulfate content without any protenaceous component were prepared from dried nori processed from Porphyra yezoensis, a red alga. Each of these preparations was applied to demonstrate adsorption or binding to the surface of oil droplets. The decrease in porphyran concentration of the aqueous phase of O/W emulsions prepared with porphyran and with toluidine blue (TB)-porphyran complex formed by adding TB to the O/W emulsions indicated ready adsorption to the surface of oil droplets. The decrease in zeta-potential of the O/W emulsions suggested that the sulfate groups of the adsorbed porphyran were oriented toward the external aqueous phase. A biomolecular interaction analysis exhibited rapid binding of porphyran to C16-alkane, probably through 3,6-anhydrogalactose. Porphyran-coated liposomes were tolerant to digestion with phospholipase D. The increased molecular weight of the porphyran preparations had an increased effect on these characteristics. The results of this study demonstrate that the emulsifying ability of porphyran is derived from the adequate adsorption to the surface of oil droplets and that porphyran could be effectively applied to stabilize liposomes.

Topics: Adsorption; Chemical Phenomena; Chemistry, Physical; Drug Stability; Emulsions; Hydrophobic and Hydrophilic Interactions; Liposomes; Phospholipase D; Porphyra; Sepharose

Arylsulfatase was purified from Sphingomonas sp. AS6330 through ionic exchange, hydrophobic- and gel-chromatographies. The purity increased 12,800-fold with approximately 19.1% yield against cell homogenate. The enzyme was a monomeric protein with apparent molecular weight of 62 kDa as determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis, and 41 kDa as determined by gel filtration. The enzyme had optimum reaction conditions for hydrolysis of sulfate ester bonds in agar and p-nitrophenyl sulfate (NPS) at pH 7.0 and 45 degrees C, with a specific activity of 3.93 and 97.2 U, respectively. The enzyme showed higher activity towards agar than other sulfated marine polysaccharides such as porphyran, fucoidan and carrageenan. The K(m) and V(max) of the enzyme for hydrolysis of NPS were 54.9 microM and 113 mM/min, respectively. With reaction of 200 g agar with 100 U arylsulfatase for 8 h at 45 degrees C, gel strength increased 2.44-fold, and 97.7% of the sulfate in the agar was hydrolyzed.

Topics: Agar; Arylsulfatases; Biotransformation; Carrageenan; Chromatography, Gel; Chromatography, Ion Exchange; Enzyme Stability; Hydrogen-Ion Concentration; Industrial Microbiology; Molecular Weight; Nitrobenzenes; Polysaccharides; Sepharose; Sphingomonas; Substrate Specificity; Temperature

Polysaccharides extracted from several red and brown seaweeds growing along the South American coasts were evaluated regarding their cytotoxic effects against HeLa cells. Sulfated galactans were isolated and purified from the red algae Bostrychia montagnei (BHW and B4 fractions, 17 and 22% SO3Na, respectively) and Porphyra columbina (PC75 fraction, 15.6% SO3Na). At maximum concentration of 80 microg/mL, these fractions promoted atypical mitoses in HeLa cells, presence of atypical nuclei and blebs. Alginic acid (160.0 microg/mL) isolated from the brown seaweed Laminaria brasiliensis (molar ratio M/G 1.2) and its M and G blocks (40.0 microg/mL; DP = 20), promoted similar morphologic alterations besides the presence of acidophilic material in the cellular cytoplasm and occurrence of multinuclear cells present in the monolayer. The polysaccharide fraction SF isolated from the brown seaweed Sargassum stenophyllum containing mainly fucose and presenting molar ratio of sulfate/sugar 1.9, promoted very accentuated morphologic modifications in HeLa cells at low concentrations. SF (2.5 microg/mL) caused significant alterations in the cellular morphology and reduction of the growth, such effects being dose dependent. At 40.0 microg/mL, accentuated decrease of the number of mitosis illustrations accompanied by an increase in the number of condensed cells, atypical nuclei, number of clusters and blebs. These results demonstrated that among the anionic polysaccharides tested, the sulfated fucans were those which caused greater cytotoxic effects in HeLa cells.

Topics: Alginates; Animals; Antineoplastic Agents; Cell Survival; Galactans; Glucuronic Acid; HeLa Cells; Hexuronic Acids; Humans; Laminaria; Polysaccharides; Seaweed; Sepharose; Structure-Activity Relationship

A suspension of low-quality dried nori processed from Porphyra yezoensis, a red alga, was autoclaved at 120 degrees C for 30 min, and from the supernatant, five preparations of porphyran of differing molecular masses and chemical compositions were obtained by preprecipitation with ethanol at stepwise-increasing concentrations of 50 and 67% followed by size-exclusion chromatography. The porphyran preparations exhibited a high emulsifying activity index and high emulsion stability over a wide range of pH and temperature and also in the presence of sodium chloride. An adequately high coefficient of correlation between the median diameter of oil droplets and their 3,6-anhydrogalactose content suggests that 3,6-anhydrogalactose could take part in emulsification with porphyran.

Topics: Excipients; Hydrogen-Ion Concentration; Rhodophyta; Sepharose

Beta-Agarase I and II were characterised by their action on agar-type polysaccharides and oligosaccharides. Beta-Agarase I, an endo-enzyme, was specific for regions containing a minimum of one unsubstituted neoagarobiose unit [3,6-anhydro-alpha-L-galactopyranosyl-(1 leads to 3)-D-galactose], hydrolysing at the reducing side of this moiety. Yaphe demonstrated that agar was degraded by this enzyme to neoagaro-oligosaccharides limited by the disaccharide but with a predominance of the tetramer [Yaphe, W. (1957) Can. J. Microbiol. 3, 987-993]. Beta-Agarase I slowly degraded neoagarohexaose but not the homologous tetrasaccharide. [1-3H]Neoagarohexaitol was cleaved to neoagarotetraose and [1-3H]neoagarobiitol. The highly substituted agar, porphyran was degraded to methylated, sulphated and unsubstituted neoagaro-oligosaccharides which were invariably terminated at the reducing end by unsubstituted neoagarobiose. The novel enzyme, beta-agarase II, was shown to be an endo-enzyme. Preliminary evidence indicated this enzyme was specific for sequences containing neoagarobiose and/or 6(1)-O-methyl-neoagarobiose. It degraded agar to neoagaro-oligosaccharides of which the disaccharide was limiting and predominant. Beta-Agarase II rapidly degraded isolated neogarotetraose and neoagarohexaose to the disaccharide. With [1-3H]neoagarohexaitol, exo-action was observed, the alditol being cleaved to neoagarobiose and [1-3H]neoagarotetraitol. Neoagarotetraitol was hydrolysed at 4% of the rate observed for the hexaitol. Porphyran was degraded to oligosaccharides, the neutral fraction comprising 24% of the starting carbohydrate. This fraction was almost exclusively disaccharides (22.4%) containing neoagarobiose (7.4%) and 6(1)-O-methyl-neoagarobiose (15%). Beta-Agarase II is probably the 'beta-neoagarotetraose hydrolase' reported by Groleau and Yaphe as an exoenzyme against neoagaro-oligosaccharides [Groleau, D. and Yaphe, W. (1977) Can. J. Microbiol. 23, 672-679].

Topics: Agar; Glycoside Hydrolases; Oligosaccharides; Pseudomonas; Sepharose; Substrate Specificity

Porphyran, a highly substituted agarose from Porphyra umbilicalis was degraded by highly purified beta-agarase I from Pseudomonas atlantica. This enzyme cleaved at the reducing side of units of beta-neoagarobiose (3,6-anhydro-alpha-L-galactopyranosyl-(1 leads to 3)-beta-D-galactopyranose). The oligosaccharides were divided into fractions of low and high molecular weight by dialysis. The permeate (23% of total starting carbohydrate) was separated by ion-exchange into neutral and anionic fractions. Gel filtration of the neutral fraction (19%) resolved two major oligosaccharides. These were shown by 13C-NMR spectroscopy to be 6(3)-O-methyl-neoagarotetraose and 6(3),6(5)-di-O-methyl-neoagarohexaose. Gel filtration of the anionic oligosaccharides (3.3%) revealed two novel monosulphated tetrasaccharides, 6-O-sulphato-alpha-L-galacto-pyranosyl-(1 leads to 3)-beta-D-galactopyranosyl-(1 leads to 4)-3,6-anhydro-alpha-L-galactopyranosyl-(1 leads to 3)-D-galactopyranose and its 6(3)-O-methylated derivative. The 13C-NMR data from the sulphated tetrasaccharides provided a novel reference which was used to characterise higher, partially sulphated fragments in the dialysis permeate. The fraction retained on dialysis (77%) had an average degree of polymerisation of 40 and was homologous with the high-molecular-weight anionic permeate. From 13C-NMR spectroscopy porphyran was found to comprise 49% sulphated disaccharide units and these were calculated to occur in stretches averaging 2.0-2.5 contiguous units.

Topics: Chemical Phenomena; Chemistry; Glycoside Hydrolases; Magnetic Resonance Spectroscopy; Oligosaccharides; Polysaccharides; Polysaccharides, Bacterial; Pseudomonas; Sepharose

Topics: Polysaccharides; Sepharose

Topics: Carbohydrate Metabolism; Enzymes; Eukaryota; Polysaccharides; Sepharose