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In this study, we purified a lectin isolated from the seeds of Dioclea bicolor (DBL) via affinity purification. Electrophoresis analysis revealed that DBL had three bands, α, ß, and γ chains, with molecular masses of approximately 29, 14, and 12 kDa, respectively. Gel filtration chromatography revealed that the native form of DBL had a molecular mass of approximately 100 kDa, indicating that it is a tetramer. Interestingly, DBL-induced hemagglutination was inhibited by several glucosides, mannosides, ampicillin, and tetracycline with minimum inhibitory concentration (MIC) values of 1.56-50 mM. Analysis of the complete amino acid sequence of DBL revealed the presence of 237 amino acids with high similarity to other Diocleinae lectins. Circular dichroism showed the prominent ß-sheet secondary structure of DBL. Furthermore, DBL structure prediction revealed a Discrete Optimized Protein Energy (DOPE) score of -26,642.69141/Normalized DOPE score of -1.84041. The DBL monomer was found to consist a ß-sandwich based on its 3D structure. Molecular docking showed the interactions between DBL and α-D-glucose, N-acetyl-D-glucosamine, α-D-mannose, α-methyl-D-mannoside, ampicillin, and tetracycline. In addition, DBL showed antimicrobial activity with an MIC of 125 µg/mL and exerted synergistic effects in combination with ampicillin and tetracycline (fractional inhibitory concentration index ≤ 0.5). Additionally, DBL significantly inhibited biofilm formation and showed no toxicity in murine fibroblasts (p < 0.05). These results suggest that DBL exhibits antimicrobial activity and works synergistically with antibiotics.
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Antibacterianos , Dioclea , Lectinas de Plantas , Antibacterianos/farmacologia , Antibacterianos/química , Antibacterianos/isolamento & purificação , Camundongos , Animais , Lectinas de Plantas/química , Lectinas de Plantas/farmacologia , Lectinas de Plantas/isolamento & purificação , Dioclea/química , Simulação de Acoplamento Molecular , Testes de Sensibilidade Microbiana , Ampicilina/farmacologia , Ampicilina/químicaRESUMO
Tomato seeds are a rich source of protein that can be utilized for various industrial food purposes. This study delves into the effects of using supercritical CO2 (scCO2) on the structure and techno-functional properties of proteins extracted from defatted tomato seeds. The defatted meal was obtained using hexane (TSMH) and scCO2 (TSMC), and proteins were extracted using water (PEWH and PEWC) and saline solution (PESH and PESC). The results showed that scCO2 treatment significantly improved the techno-functional properties of protein extracts, such as oil-holding capacity and foaming capacity (especially for PEWC). Moreover, emulsifying capacity and stability were enhanced for PEWC and PESC, ranging between 4.8 and 46.7% and 11.3 and 96.3%, respectively. This was made possible by the changes in helix structure content induced by scCO2 treatment, which increased for PEWC (5.2%) and decreased for PESC (8.0%). Additionally, 2D electrophoresis revealed that scCO2 hydrolyzed alkaline proteins in the extracts. These findings demonstrate the potential of scCO2 treatment in producing modified proteins for food applications.
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A type of high molecular weight bioactive polymers called exopolysaccharides (EPS) are produced by thermophiles, the extremophilic microbes that thrive in acidic environmental conditions of hot springs with excessively warm temperatures. Over time, EPS became important as natural biotechnological additives because of their noncytotoxic, emulsifying, antioxidant, or immunostimulant activities. In this article, we unravelled a new EPS produced by Staphylococcus sp. BSP3 from an acidic (pH 6.03) San Pedro hot spring (38.1 °C) located in the central Andean mountains in Chile. Several physicochemical techniques were performed to characterize the EPS structure including Scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS), Atomic Force Microscopy (AFM), High-Performance Liquid Chromatography (HPLC), Gel permeation chromatography (GPC), Fourier Transform Infrared Spectroscopy (FTIR), 1D Nuclear Magnetic Resonance (NMR), and Thermogravimetric analysis (TGA). It was confirmed that the amorphous surface of the BSP3 EPS, composed of rough pillar-like nanostructures, is evenly distributed. The main EPS monosaccharide constituents were mannose (72%), glucose (24%) and galactose (4%). Also, it is a medium molecular weight (43.7 kDa) heteropolysaccharide. NMR spectroscopy demonstrated the presence of a [â 6)-âº-D-Manp-(1 â 6)-âº-D-Manp-(1 â] backbone 2-O substituted with 1-âº-D-Manp. A high thermal stability of EPS (287 °C) was confirmed by TGA analysis. Emulsification, antioxidant, flocculation, water-holding (WHC), and oil-holding (OHC) capacities are also studied for biotechnological industry applications. The results demonstrated that BSP3 EPS could be used as a biodegradable material for different purposes, like flocculation and natural additives in product formulation.
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Althaea officinalis Linn. (AO) is a widely distributed herbaceous plant with a long history of medicinal and food functions in Europe and Western Asia. Althaea officinalis polysaccharide (AOP), as one of the main components and a crucial bioactive substance of AO, has a variety of pharmacological activities, including antitussive, antioxidant, antibacterial, anticancer, wound healing, immunomodulatory, and infertility therapy effects. Many polysaccharides have been successfully obtained in the last five decades from AO. However, there is currently no review available concerning AOP. Considering the importance of AOP for biological study and drug discovery, the present review aims to systematically summarize the recent major studies on extraction and purification methods of polysaccharides from different AO parts (seeds, roots, leaves and flowers), as well as the characterization of their chemical structure, biological activity, structure-activity relationship, and the application of AOP in different fields. Meanwhile, the shortcomings of AOP research are further discussed in detail, and new valuable insights for future AOP research as therapeutic agents and functional foods are proposed.
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Althaea , Antitussígenos , Althaea/química , Polissacarídeos/química , Antioxidantes/química , Extratos Vegetais/farmacologia , Antitussígenos/farmacologiaRESUMO
In the present work, the chemical synthesis of AgPt nanoalloys is reported by the polyol method using polyvinylpyrrolidone (PVP) as a surfactant and a heterogeneous nucleation approach. Nanoparticles with different atomic compositions of the Ag and Pt elements (1:1 and 1:3) were synthesized by adjusting the molar ratios of the precursors. The physicochemical and microstructural characterization was initially performed using the UV-Vis technique to determine the presence of nanoparticles in suspension. Then, the morphology, size, and atomic structure were determined using XRD, SEM, and HAADF-STEM techniques, confirming the formation of a well-defined crystalline structure and homogeneous nanoalloy with an average particle size of less than 10 nm. Finally, the cyclic voltammetry technique evaluated the electrochemical activity of bimetallic AgPt nanoparticles supported on Vulcan XC-72 carbon for the ethanol oxidation reaction in an alkaline medium. Chronoamperometry and accelerated electrochemical degradation tests were performed to determine their stability and long-term durability. The synthesized AgPt (1:3)/C electrocatalyst presented significative catalytic activity and superior durability due to the introduction of Ag that weakens the chemisorption of the carbonaceous species. Thus, it could be an attractive candidate for cost-effective ethanol oxidation compared to commercial Pt/C.
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The current study investigated the effects of two different pretreatments (NaOH and alkaline surfactant assisted) on the chemical, morphological and enzymatic saccharification of pineapple leaf fiber (PALF). Results showed that both pretreatments significantly reduced lignin content of the biomass, achieving a 69.6 and a 76.3% reduction for NaOH and surfactant pretreated materials, respectively. SEM, CLSM and FTIR-ATR techniques were used to evaluate morphological changes in the fibers after pretreatments. Images obtained revealed cellulose exposure and lignin redistribution in the pretreated fibers. Surfactant pretreated material provided the best results after enzymatic hydrolysis compared to NaOH and untreated PALF. A final enzymatic hydrolysis yield of 81.8% was obtained after a 24 h process using surfactant pretreated fibers, in comparison to 75.9 and 45.1% yields for NaOH pretreated material and raw fibers, respectively. Nowadays, the use of agricultural residues for high added value products is of great importance for sustainable development. This work specifically studied an effective and green approach for lignin removal and enzymatic hydrolysis from pineapple leaf fiber that is an abundant waste in Costa Rica and an interesting feedstock for biorefinery processes design.
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Álcalis , Ananas , Fibras na Dieta , Hidrólise , Lignina/química , Folhas de Planta , Hidróxido de Sódio/química , TensoativosRESUMO
This study aimed to isolate, characterize chemical-structurally and evaluate the effects of polysaccharides from Caesalpinia (Libidibia) ferrea stem barks in the haemostatic system. The deproteinated-polysaccharide extract (PE-Cf) after being fractionated by ion exchange chromatography-DEAE-cellulose resulted in three fractions (FI, FII, FIII) containing total carbohydrates (14.3-38%), including uronic acid (5-16%), and polyphenols (0.94-1.7 mg/g GAE). The polysaccharide fractions presented polydisperse profile in polyacrylamide gel electrophoresis (detected by Stains-All) and molecular masses (9.5 × 104 Da-1.5 × 105 Da) identified by gel permeation chromatography. FT-IR showed absorption bands (1630 cm-1, 1396-1331 cm-1), indicative of uronic acid, and a band at 1071 cm-1, typical of COO- groups of galacturonic acid. The NMR spectra of C. ferrea polysaccharides revealed a central core composed mainly by 5-linked α-Araf and minority components as α-Rhap and α-GalAp. UV spectra of fractions revealed discrete shoulders at 269-275 nm, characteristic of polyphenolic compounds. In vitro, polysaccharides inhibited the intrinsic and/or common coagulation pathway (aPTT test) (2.0-3.7 fold) and the platelet aggregation induced by 3 µM adenosine diphosphate (25-48%) and 5 µg/mL collagen (24%), but not that induced by arachidonic acid. In vivo, the polysaccharides inhibited (36-69%) venous thrombosis induced by hypercoagulability and stasis, showing discrete hemorrhagic effect. In conclusion, the polysaccharides of C. ferrea barks, containing arabinose, galactose, rhamnose and uronic acid, possess anticoagulant, antiplatelet and antithrombotic properties of low hemorrhagic risk, suggesting potential applicability in thromboembolic disorders.
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Caesalpinia/metabolismo , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Animais , Anticoagulantes/química , Coagulação Sanguínea/efeitos dos fármacos , Fibrinolíticos/química , Humanos , Tempo de Tromboplastina Parcial , Casca de Planta/química , Extratos Vegetais/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Polifenóis/farmacologia , Ratos , Ratos Wistar , Trombose VenosaRESUMO
Schistosomiasis is a neglected tropical disease that affects more than 250 million people worldwide. The only drug available for its treatment undergoes first-pass hepatic metabolism and is not capable of preventing reinfection, which makes the search of new therapies urgently needed. Due to the essential role of fumarases in metabolism, these enzymes represent potential targets for developing novel schistosomiasis treatments. Here, we evaluate the expression profiles for class I and class II fumarases from Schistosoma mansoni (SmFHI and SmFHII, respectively), and report the complete characterization of SmFHII. The first SmFHII structure in complex with L-malate was determined at 1.85 Å resolution. The significant thermoshift observed for SmFHII in the presence of identified ligands makes the differential scanning fluorimetry an adequate technique for ligand screening. A complete kinetic characterization of SmFHII was performed, and comparison with the human fumarase (HsFH) revealed differences regarding the turnover number (kcat). Structural characterization allowed us to identify differences between SmFHII and HsFH that could be explored to design new selective inhibitors. This work represents the very first step towards validate the fumarases as drug targets to treat schistosomiasis. Our results provide the structural basis to rational search for selective ligands.
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Fumarato Hidratase/farmacologia , Schistosoma mansoni/enzimologia , Esquistossomose mansoni/tratamento farmacológico , Animais , Feminino , Fumarato Hidratase/metabolismo , Cinética , Ligantes , Masculino , Camundongos , Schistosoma mansoni/metabolismo , Esquistossomose/tratamento farmacológico , Esquistossomose/metabolismo , Esquistossomose mansoni/metabolismoRESUMO
Some physical properties of spider silks, including mechanical strength and toughness, have been studied in many laboratories worldwide. Given that this silk is organic in nature, composed of protein, and has similar properties to metal wires or polymers, it has the potential for application in medicine, nanoelectronics, and other related areas. In this study, we worked on spider silk from the Nephila clavipes species collected from the wild and kept it in the nursery of the Autonomous University of the West, Cali, Colombia, to determine its physical, thermal, and mechanical properties, seeking possible applications in the medical and industrial sectors and comparing the material properties of the silk from the species from southwestern Colombia with those of the previously studied species from other regions. The mechanical characterization of the material was performed using a universal testing machine; thermal behavior was captured by a thermogravimetric analysis, differential scanning calorimetry, and mass spectrometry; and structural characterization was performed using diffraction X-rays. The results of the thermal characterization demonstrate that the spider silk loses 10 % of water content at 150 °C with significant changes at 400 °C, while the mechanical characterization indicates that the spider silk is much tougher than Kevlar 49 and Nylon 6 since it is capable of absorbing more energy before rupture.
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Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most frequent human enzymopathy, affecting over 400 million people globally. Worldwide, 217 mutations have been reported at the genetic level, and only 19 have been found in Mexico. The objective of this work was to contribute to the knowledge of the function and structure of three single natural variants (G6PD A+, G6PD San Luis Potosi, and G6PD Guadalajara) and a double mutant (G6PD Mount Sinai), each localized in a different region of the three-dimensional (3D) structure. In the functional characterization of the mutants, we observed a decrease in specific activity, protein expression and purification, catalytic efficiency, and substrate affinity in comparison with wild-type (WT) G6PD. Moreover, the analysis of the effect of all mutations on the structural stability showed that its presence increases denaturation and lability with temperature and it is more sensible to trypsin digestion protease and guanidine hydrochloride compared with WT G6PD. This could be explained by accelerated degradation of the variant enzymes due to reduced stability of the protein, as is shown in patients with G6PD deficiency.
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Deficiência de Glucosefosfato Desidrogenase/enzimologia , Deficiência de Glucosefosfato Desidrogenase/genética , Glucosefosfato Desidrogenase/química , Glucosefosfato Desidrogenase/metabolismo , Naftalenossulfonato de Anilina/química , Catálise , Dicroísmo Circular , Glucosefosfato Desidrogenase/genética , Glucosefosfato Desidrogenase/isolamento & purificação , Deficiência de Glucosefosfato Desidrogenase/metabolismo , Guanidina , Humanos , Cinética , México , Modelos Moleculares , Mutagênese Sítio-Dirigida , Mutação , Estabilidade Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Software , Temperatura , Tripsina/químicaRESUMO
Phycobiliproteins (PBPs) are colored fluorescent proteins present in cyanobacteria, red alga, and cryptophyta. These proteins have many potential uses in biotechnology going from food colorants to medical applications. Allophycocyanin, the simplest PBP, is a heterodimer of αß subunits that oligomerizes as a trimer (αß)3 . Each subunit contains a phycocyanobilin, bound to a cysteine residue, which is responsible for its spectroscopic properties. In this article, we are reporting the expression of recombinant allophycocyanin (rAPC) from the eukaryotic red algae Agarophyton chilensis in Escherichia coli, using prokaryotic accessory enzymes to obtain a fully functional rAPC. Three duet vectors were used to include coding sequences of α and ß subunits from A. chilensis and accessorial enzymes (heterodimeric lyase cpc S/U, heme oxygenase 1, phycocyanobilin oxidoreductase) from cyanobacteria Arthrospira maxima. rAPC was purified using several chromatographic steps. The characterization of the pure rAPC indicates very similar spectroscopic properties, λmaxAbs , λmaxEm , fluorescence lifetime, and chromophorylation degree, with native allophycocyanin (nAPC) from A. chilensis. This method, to produce high-quality recombinant allophycocyanin, can be used to express and characterize other macroalga phycobiliproteins, to be used for biotechnological or biomedical purposes.
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Eucariotos/genética , Ficocianina/biossíntese , Ficocianina/genética , Células Procarióticas/enzimologia , Proteínas Recombinantes , Eletroforese em Gel de Poliacrilamida , Expressão Gênica , Vetores Genéticos/genética , Peso Molecular , Ficocianina/isolamento & purificação , Análise EspectralRESUMO
This article presents the data on α-Fe2O3 nanoparticles synthesized via Pechini method using iron(III) oxide precursor from steel industry. It is important to highlight the added value that is given to an industrial waste. The samples were characterized by thermal analysis (DTA, TG), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The TG showed three mass changes, whereas DTA resulted in three anomalies. X-ray diffraction pattern of the samples disclosed rhombohedral structure characteristic of the nanocrystalline α-Fe2O3 phase. The crystallite size was estimated for each thermal treatment. Fourier transform infrared spectroscopy confirms the phase purity of prepared nanoparticles. A detailed study on the local structure of the samples was carry out in the region of 800 and 400 cm-1, where the associated bands of Fe-O bonds are presents. The data have not been reported nor discussed for now.
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Lomentospora prolificans is an emerging opportunistic fungus with a high resistance to antifungal agents and it can cause localized infections in immunocompetent patients and disseminated infections with a high mortality rate in immunosuppressed patients. Glucosylceramides (GlcCer) are synthetized in the majority of known fungal pathogens. They are bioactive molecules presenting different functions, such as involvement in fungal growth and morphological transitions in several fungi. The elucidation of the primary structure of the fungal surface glycoconjugates could contribute for the understanding of the mechanisms of pathogenicity. In this work, GlcCer species were isolated from mycelium and conidia forms of L. prolificans and their chemical structures were elucidated by mass spectrometry (ESI-MS). GlcCer purified from both forms presented a major species at m/z 750 that corresponds to N-2-hydroxyhexadecanoyl-1-ß-D-glucopyranosyl-9-methyl-4,8-sphingadienine. Monoclonal antibodies against GlcCer could recognize L. prolificans GlcCer species from mycelium and conidia, suggesting a conserved epitope in fungal GlcCer. In addition, in vivo assays showed that purified GlcCer species from both forms was able to induce a high secretion of pro-inflammatory cytokines by splenocytes. GlcCer species also promote the recruitment of polymorphonuclear, eosinophils, small peritoneal macrophage (SPM) and mononuclear cells to the peritoneal cavity. GlcCer species were also able to induce the oxidative burst by peritoneal macrophages with NO and superoxide radicals production, and to increase the killing of L. prolificans conidia by peritoneal macrophages. These results indicate that GlcCer species from L. prolificans are a potent immune response activator.
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As part of the ongoing efforts in discovering potentially bioactive natural products from medicinal plants, the present study was conducted to isolate a new complex triterpenoid saponin from the barks of Albizia lebbeck. It was isolated by using chromatographic methods and its structural elucidation was performed using detailed analyses of 1H and 13C NMR spectra including 2D-NMR (COSY, TOCSY, HSQC and HMBC) spectroscopic techniques, high-resolution electrospray ionization mass spectrometry (HRESIMS) analysis and chemical conversions. Its structure was established as 21-[[(2E,6S)-6-[6-deoxy-4-O-[(2E,6S)-6-hydroxy-2-(hydroxymethyl)-6-methyl-1-oxo-2,7-octadienyl]-[(ß-d-glucopyranosyl)oxy]-2-(hydroxymethyl)-6-methyl-1-oxo-2,7-octadienyl]-[(ß-d-glucopyranosyl)oxy]-2,6-dimethyl-1-oxo-2,7-octadienyl]oxy]-16-hydroxy-3-[[O-ß-d-xylopyranosyl-(1â¯ââ¯2)-O-α-l-arabinopyranosyl-(1â¯ââ¯6)-2-(acetylamino)-2-deoxy-ß-d-glucopyranosyl]oxy]-(3ß,16α,21ß)-olean-12-en-28-oic acid O-α-l-arabinofuranosyl-(1â¯ââ¯4)-O-[ß-d-glucopyranosyl-(1â¯ââ¯3)]-O-6-deoxy-α-l-mannopyranosyl-(1â¯ââ¯2)-ß-d-glucopyranosyl ester (1). Additionally, this study aimed to investigate the permeability property of 1, its activity on membrane integrity and supramolecular interactions with cellular constituents using in vitro experimental models.
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Albizzia/química , Membrana Celular/efeitos dos fármacos , Saponinas/química , Triterpenos/química , Animais , Eritrócitos/efeitos dos fármacos , Humanos , Espectroscopia de Ressonância Magnética , Masculino , Camundongos , Estrutura Molecular , Permeabilidade , Saponinas/farmacocinética , Espectrometria de Massas por Ionização por Electrospray , Triterpenos/farmacocinéticaRESUMO
Differences in anomericity and in the branching degree of glucans lead to characteristic intermolecular association that influences their solubility in water or other solvents. A simple purification approach, based on the glucan solubility in aq. 0.1 M NaOH solution, was applied for the separation of mixed water-insoluble α-D-glucans from ß-D-glucans extracted from fruiting bodies of Fomitopsis betulina, which is an underexploited medicinal mushroom. The results indicated that the ß-D-glucan is constituted by (1â3)-linked ß-D-Glcp units substituted at O-6 by non-reducing ß-D-Glcp and (1â6)-linked ß-D-Glcp units, while the α-D-glucan has a linear (1â3)-linked glucan structure. Thus, the 0.1 M NaOH treatment proved to be a simple, efficient and low-cost purification method for separation of water-insoluble glucans with different anomeric configurations and degree of branching that were interacting by intermolecular forces.
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Coriolaceae/química , Glucanos/química , Água/química , Sequência de Carboidratos , Solubilidade , EstereoisomerismoRESUMO
Cell wall polysaccharides from ponkan peel were investigated with the aim of gain knowledge about their potential for different applications and the use of ponkan peel as raw material for pectin extraction. The plant material was defatted using MeOH:CHCl3, pretreated with DMSO and then subjected to sequential extractions with cold and hot water, ammonium oxalate, HCl, Na2CO3, 2â¯M and 4â¯M NaOH in order to obtain polysaccharides. The polysaccharide fractions were analyzed by chemical, chromatographic and spectroscopic methods Cold and hot water-soluble pectins contained higher amounts of GalA and higher degrees of methyl-esterification (DM) than ammonium oxalate and HCl fractions. Na2CO3 extraction provided non-esterified arabinose-rich pectins which formed gel in a dialysis step. NaOH solubilized hemicelluloses, composed mainly of xyloglucans, galactomannans and galactoglucomannans. The water-soluble fraction (WSP) was purified using α-amylase and amyloglucosidase and gave rise to the subfraction named α-WSP. The α-WSP was a pectin composed of HG and RG-I domains containing side chains of arabinans and short-chains of galactans, with low DM (39.4%) and Mw of 1.615â¯×â¯105â¯g/mol.
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A central composite experimental design was used to evaluate the influence of pH, extraction time and liquid:solid ratio on the yield and uronic acid content of the pectin from ponkan peel. The response surface methodology showed that the yield is positively influenced by lower pHs, longer extraction times and higher liquid:solid ratio, whereas the uronic acid content decreases with increasing extraction time. The conditions that resulted in the highest yield and highest uronic acid content were defined as pHâ¯1.6, extraction time of 100â¯min and liquid:solid ratio of 36â¯mL/g. The pectin obtained under these conditions (PPOP) had an experimental yield of 25.6%, below the predicted theoretical value despite the good fit of the model (R2â¯=â¯0.96) and the galacturonic acid content was 84.5%, in close agreement with the predicted theoretical value. PPOP was composed mainly of a homogalacturonan with degree of methyl esterification of 85.7% and a rhamnogalacturonan I region mainly branched by galactans. In addition, PPOP had a very low degree of acetylation (0.1%) and average molar mass of 80,650â¯g/mol, determined by light scattering. The results showed that ponkan peel may be used as a source of citrus pectin in the regions where this species is cultivated.
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Citrus/química , Pectinas/química , Pectinas/isolamento & purificação , Concentração de Íons de Hidrogênio , Pressão , Espectroscopia de Prótons por Ressonância Magnética , Ácidos Urônicos/análiseRESUMO
Abstract Ximenia americana L., Olacaceae, barks are utilized in folk medicine as analgesic and anti-inflammatory. The objective was to evaluate the toxicity and antinociceptive effect of polysaccharides rich fractions from X. americana barks. The fractions were obtained by extraction with NaOH, followed by precipitation with ethanol and fractionation by ion exchange chromatography. They were administered i.v. or p.o. before nociception tests (writhing, formalin, carragenan-induced hypernociception, hot plate), or during 14 days for toxicity assay. The total polysaccharides fraction (TPL-Xa: 8.1% yield) presented 43% carbohydrate (21% uronic acid) and resulted in two main fractions after chromatography (FI: 12%, FII: 22% yield). FII showed better homogeneity/purity, content of 44% carbohydrate, including 39% uronic acid, arabinose and galactose as major monosaccharides, and infrared spectra with peaks in carbohydrate range for COO- groups of uronic acid. TPL-Xa (10 mg/kg) and FII (0.1 and 1 mg/kg) presented inhibitory effect in behavior tests that evaluate nociception induced by chemical and mechanical, but not thermal stimuli. TPL-Xa did not alter parameters of systemic toxicity. In conclusion, polysaccharides rich fractions of X. americana barks inhibit peripheral inflammatory nociception, being well tolerated by animals.
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A crude fraction (SCW) was extracted with cold water from Pholiota nameko and showed mannose (24.1%), galactose (44.9%) and glucose (31%). Purification procedures resulted in a polysaccharide fraction (bG-PN), that showed only glucose. NMR and methylation analyses of bG-PN indicated a ß-d-glucan-(1â3)-linked, substituted at O-6 by ß-d-Glcp or (1â6)-linked ß-d-Glcp side chains. Rheological studies of crude and purified fractions at the same concentration showed similar shear-thinning behavior and gel-like structure which indicates no need to isolate the polymer to achieve some desirable rheological properties. SCW (at 1% and 2%) and bG-PN (at 2%) presented thermal stability during heating and cooling, suggesting that the physical structure of gels (SCW and bG-PN at 2%) and viscoelastic fluid (SCW at 1%) formed were not altered in the tested temperature range. Our results suggest that P. nameko ß-d-glucans can be applied in different food preparations as thickener or gelling agents modifying their rheological properties.
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beta-Glucanas/química , Pholiota , Reologia , Substâncias Viscoelásticas/química , Substâncias Viscoelásticas/isolamento & purificação , beta-Glucanas/isolamento & purificaçãoRESUMO
ABSTRACT Gracilariopsis lemaneiformis is a type of red alga that contains seaweed polysaccharide agar. In this study, a novel non-agar seaweed polysaccharide fraction named GCP (short of crude polysaccharide obtained from Gracilariopsis lemaneiformis) was isolated from Gracilariopsis lemaneiformis. Structural analysis showed that GCP shows triple helical chain conformation when dissolved in water and has many branches and long side chains. Also, 1→3 linkage is the major linkage and the sugar structures are galactopyranose configurations linked by β-type glycosidic linkages. Two macromolecular substance fractions (GCP-1 and GCP-2) were purified by DEAE Sepharose Fast Flow column chromatography. Moreover, a splenocyte damage assay and splenocyte proliferation assay were used to analyse the bioactivities of GCP, GCP-1 and GCP-2. It was demonstrated that polysaccharides could protect splenocyte damaged by H2O2; GCP-2 shows a greatest protection rate, that is, 92.8%, which significantly enhanced the splenocyte proliferation, and GCP showed the highest proliferation rate, 9.30%. The results suggested that this type of novel non-agar polysaccharide displayed remarkable antioxidant and immunomodulatory activities and early alkali treatment could decrease the activities. It may represent a potential material for health food and clinical medicines.