Enhancement of the Antioxidant and Antimicrobial Activities of Porphyran through Chemical Modification with Tyrosine Derivatives.

The chemical modification of porphyran hydrocolloid is attempted, with the objective of enhancing its antioxidant and antimicrobial activities. Sulfated galactan porphyran is obtained from commercial samples of the red algae Porphyra dioica using Soxhlet extraction with water at 100 °C and precipitation with isopropyl alcohol. The extracted porphyran is then treated with modified L-tyrosines in aqueous medium in the presence of NaOH, at ca. 70 °C. The modified tyrosines L1 and L2 are prepared through a Mannich reaction with either thymol or 2,4-di-tert-butylphenol, respectively. While the reaction with 2,4-di-tert-butylphenol yields the expected tyrosine derivative, a mixture of products is obtained with thymol. The resulting polysaccharides are structurally characterized and the respective antioxidant and antimicrobial activities are determined. Porphyran treated with the N-(2-hydroxy-3,5-di-tert-butyl-benzyl)-L-tyrosine derivative, POR-L2, presents a noticeable superior radical scavenging and antioxidant activity compared to native porphyran, POR. Furthermore, it exhibited some antimicrobial activity against S. aureus. The surface morphology of films prepared by casting with native and modified porphyrans is studied by SEM/EDS. Both POR and POR-L2 present potential applicability in the production of films and washable coatings for food packaging with improved protecting characteristics.

Porphyran and oligo-porphyran originating from red algae Porphyra: Preparation, biological activities, and potential applications.

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 ß-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.

Optimization of Porphyran Extraction from Pyropia yezoensis by Response Surface Methodology and Its Lipid-Lowering Effects.

Macroalgae polysaccharides are phytochemicals that are beneficial to human health. In this study, response surface methodology was applied to optimize the extraction procedure of Pyropia yezoensis porphyran (PYP). The optimum extraction parameters were: 100 °C (temperature), 120 min (time), and 29.32 mL/g (liquid-solid ratio), and the maximum yield of PYP was 22.15 ± 0.55%. The physicochemical characteristics of PPYP, purified from PYP, were analyzed, along with its lipid-lowering effect, using HepG2 cells and Drosophila melanogaster larvae. PPYP was a ß-type sulfated hetero-rhamno-galactan-pyranose with a molecular weight of 151.6 kDa and a rhamnose-to-galactose molar ratio of 1:5.3. The results demonstrated that PPYP significantly reduced the triglyceride content in palmitic acid (PA)-induced HepG2 cells and high-sucrose-fed D. melanogaster larvae by regulating the expression of lipid metabolism-related genes, reducing lipogenesis and increasing fatty acid ß-oxidation. To summarize, PPYP can lower lipid levels in HepG2 cells and larval fat body (the functional homolog tissue of the human liver), suggesting that PPYP may be administered as a potential marine lipid-lowering drug.

Physicochemical properties and potential beneficial effects of porphyran from Porphyra haitanensis on intestinal epithelial cells.

This study examined the beneficial effects of porphyran from Porphyra haitanensis (PHP) on intestinal epithelial cells, in terms of cell proliferation and migration and elucidated the potential molecular mechanism of action of PHP. Purified PHP is a homogenous polysaccharide with a molecular weight of 2.01 × 105 Da, intrinsic viscosity [η] of 463.76 mL/g, and radius of gyration of 61.2 nm. When the intestinal epithelial wound healing activity of PHP was investigated in vitro using the IEC-6 cell line (intestinal epithelial cells-6), it was found that PHP could promote cell migration and proliferation. PHP enhanced the protein expression of cell division control protein 42, paxillin, and focal adhesion kinase, which suggest that PHP might modulate the expression of these proteins to improve intestinal epithelial healing. Thus, this study indicated that PHP could serve as a potential source of functional food constituents for intestinal epithelial protection and restoration.

Antitumor bioactivity of porphyran extracted from Pyropia yezoensis Chonsoo2 on human cancer cell lines.

BACKGROUND: 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. RESULTS: 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. CONCLUSION: 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.

Structure and Bioactivities of Porphyrans and Oligoporphyrans.

BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Evaluation of Prebiotic Potential of Three Marine Algae Oligosaccharides from Enzymatic Hydrolysis.

Alginate oligosaccharides (AlgO), agarose oligosaccharides (AO), and κ-carrageenan oligosaccharides (KCO) were obtained by specific enzymatic hydrolysis method. The molecular weight distributions of the three oligosaccharides were 1.0⁻5.0 kDa, 0.4⁻1.4 kDa, and 1.0⁻7.0 kDa, respectively. The culture medium was supplemented with the three oligosaccharides and fermented by pig fecal microbiota in vitro, for 24 h. Each oligosaccharide was capable of increasing the concentration of short-chain fatty acids (SCFAs), especially butyric acid, and altering the microbiota composition. Linear discriminant analysis effect size (LEfSe) analysis results showed that the opportunistic pathogenic bacteria Escherichia, Shigella, and Peptoniphilus, were significantly decreased in AlgO supplemented medium. AO could improve the gut microbiota composition by enriching the abundance of Ruminococcaceae, Coprococcus, Roseburia, and Faecalibacterium. Besides, KCO could increase the abundance of SCFA microbial producers and opportunistic pathogenic flora. Therefore, these results indicate that AlgO and AO can be used as gut microbial regulators and can potentially improve animal/human gastrointestinal health and prevent gut disease, whereas the physiological function of KCO needs further evaluation.

Anionic carboxymethylagarose-based pH-responsive smart superabsorbent hydrogels for controlled release of anticancer drug.

Herein, we demonstrate the preparation of superabsorbent hydrogel (CMA-g-PAm) materials using anionic and cold water soluble carboxymethyagarose (CMA, a seaweed polysaccharide-based derivative) with polyacrylamide (PAm) through rapid microwave assisted grafting technique. The successful fabrication of CMA-g-PAm was verified by FT-IR, SEM, XRD, TGA and CHN analyzer. Effects of initiator, crosslinker, and monomer on the swelling behavior and grafting parameters have been thoroughly investigated. Moreover, the swelling behavior of the resulting hydrogels was systematically studied, and the results suggested they exhibit excellent pH and salt responsive behavior. The drug delivery application of thus fabricated hydrogel was further evaluated using doxorubicin (Dox) as a model drug to explore its possible applications. Release study results revealed that Dox release was significantly accelerated with decrease in pH from 7.4 to 5.0. Toxicity assays confirmed that the blank hydrogels had negligible toxicity to normal cells (VERO), whereas the Dox-loaded hydrogels remained high in cytotoxicity for A549 and Hep-G2 cancer cells. All of these attributes implied that the new proposed hydrogel (HK11) serves as potential drug delivery platforms for cancer therapy.

Protective effect of porphyran isolated from discolored nori (Porphyra yezoensis) on lipopolysaccharide-induced endotoxin shock in mice.

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.

Antioxidant and anti-inflammatory activities of porphyran isolated from discolored nori (Porphyra yezoensis).

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.

Alkanethiol-functionalized terahertz metamaterial as label-free, highly-sensitive and specific biosensor.

Specific biorecognition is essential for many biological processes, for which highly sensitive and label-free biosensors are strongly demanded. The recently developed metamaterials are a potential choice for biosensing due to their exotic properties. In the current work, a label-free and specific sensor for streptavidin-agarose (SA) was fabricated based on terahertz metamaterial functionlized by octadecanthiols and biotins. Both low and high frequency resonant modes from the metamaterials are found applicable for the detection of SA, and a redshift up to 6.76 GHz for the high frequency mode was measured in the undiluted commercial solution. The low frequency mode is attributed to inductor-capacitor (LC) oscillation, while the high frequency mode originates from the plasmonic dipole oscillator, both of which are highly sensitive to the micro-environment change. Adsorption of SA of different concentrations causes different redshifts, and the replacement of high refractive-index substrate with low refractive-index substrate can efficiently promote the sensitivity, well agreeing with the numerical simulation. Moreover, for a particular biomolecule, the sensitivity can be further improved by optimizing the metamaterial design. This method might be very helpful for desirable biorecognition in biology, medicine, and drug industry.

Structural characterization of natural ideal 6-O-sulfated agarose from red alga Gloiopeltis furcata.

A charge and size uniform polysaccharide GW2M was extracted with cold water from red alga Gloiopeltis furcata and purified by strong anion ion-exchange and gel permeation chromatography. Its chemical structure was identified by methylation, (1)H-(1)H COSY, (1)H-(13)C HMQC and (1)H-(13)C HMBC techniques. The experimental data showed that GW2M was composed of galactose (40.3%), 3,6-anhydro-galactose (34.1%) and sulfate (24.8%) with an average molecular mass of 20.6 kDa. The results proved GW2M was a linear repeating sequence of alternating (1→3)-linked 6-O-sulfated-ß-d-galactose (G6S) and (1→4)-linked 3,6-anhydro-α-l-galactose (A) which made it to be an ideal 6-O-sulfated-agarose. The sequences of serial oligosaccharides prepared by mild acid and reductive acid hydrolysis from GW2M were confirmed using electrospray collision induced dissociation tandem mass spectrometry (ES-CID-MS/MS) technique.

[Modification of seaweed polysaccharide-agarose and its application as skin dressing (III)--skin regeneration with agarose grafting hyaluronic acid sponge].

In this paper, a kind of skin dressing, agarose- grafting- hyaluronic acid (Ag-g-HA) sponge was applied to test the modified agarose based scaffold for skin regeneration. The bFGF loading agarose-grafting hyaluronan scaffold had homogenous porosities, and the loaded bFGF was bioactive in 2 weeks. The Ag-g-HA sponge was applied into skin of mice, and it was found that the dressing promoted skin regeneration and no infection and leakage in lesion site took place. H&E staining results showed that the repaired skin was similar to autologous skin. These demonstrate that Ag-g-HA sponge has a promise in skin regeneration.

Preparation of the different derivatives of the low-molecular-weight porphyran from Porphyra haitanensis and their antioxidant activities in vitro.

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.

Isolation of porphyran-degrading marine microorganisms from the surface of red alga, Porphyra yezoensis.

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.

Comparison of linear gradient and displacement separations in ion-exchange systems.

The linear gradient mode of chromatography is the most widely employed mode of operation in ion-exchange chromatographic separations. However, in recent years, the displacement mode has received considerable attention because of its promise of high throughput and high resolution. To enable a comparison of these two modes of chromatography, it is essential to identify the optimum operating conditions for each. We employed an iterative algorithm to carry out the necessary optimization. The Steric Mass Action model of ion-exchange chromatography is used in concert with the solid-film linear-driving force model to describe the chromatographic behavior of the solutes in these systems. The performances of displacement and gradient modes of chromatography are compared for different types of separation problems. It turns out that for "easy" separations, both the modes are equally effective. However, for challenging separations, the displacement mode is superior to the gradient mode. Our results shed significant light on the performance of gradient and displacement modes in protein ion-exchange systems.

Cytotoxic effects against HeLa cells of polysaccharides from seaweeds.

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.

Emulsifying ability of porphyran prepared from dried nori, Porphyra yezoensis, a red alga.

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.

[Preparation of agar and agarose from the red algae Ahnfeltia tobuchiensis]. / Poluchenie agara i agarozy iz krasnoi vodorosli Ahnfeltia tobuchiensis.

A simple and inexpensive procedure of agar and agarose production from the red alga Ahnfeltia tobuchiensis was developed, which needs no additional chromatographic purification.

Agarose-carrageenan hybrid polysaccharides from Lomentaria catenata.

Two polysaccharide fractions, PS1 and PS2 from Lomentaria catenata consisted of D-Gal, L-Gal, D-Glc, D-Xyl, D-GlcA and sulphate. Partial hydrolysis led to the isolation and identification of oligosaccharides indicating the co-existence of an agarose and carrageenan backbone structure, in which D-Glc and D-GlcA residues occur as single units branching at O-3 of -->4)alpha-D-Gal(1--> and O-4 of -->3)beta-D-Gal(1-->, respectively.