Bioactive polysaccharides from red seaweed as potent food supplements: a systematic review of their extraction, purification, and biological activities.

Most marine macroalgae such as red seaweeds are potential alternative sources of useful bioactive compounds. Beside serving as food source, recent studies have shown that red seaweeds are rich sources of bioactive polysaccharides. Red seaweed polysaccharides (RSPs) have various physiological and biological activities, which allow them to be used as immunomodulators, anti-obesity agents, and prebiotic ingredients. Lack of summary information and human clinical trials on the various polysaccharides from red seaweeds, however limits industrial-scale utilization of RSPs in functional foods. This review summarizes recent information on the approaches used for RSPs extraction and purification, mechanistic investigations of their biological activities, and related molecular principles behind their purported ability to prevent diseases. The information here also provides a theoretical foundation for further research into the structure and mechanism of action of RSPs and their potential applications in functional foods.

Restitution of epithelial cells during intestinal mucosal wound healing: The effect of a polysaccharide from the sclerotium of Lignosus rhinocerotis (Cooke) Ryvarden.

ETHNOPHARMACOLOGICAL RELEVANCE: Lignosus rhinocerotis (Cooke) Ryvarden cultivar TM02, also known as tiger's milk mushroom, is regarded as important folk medicine in Malaysia, while is used for the treatment of liver cancer, chronic hepatitis, gastric ulcer in traditional Chinese medicine. However, there is no compilation of scientific evidence that its protection for gastric, and no attempts have been made to understand how polysaccharides in Lignosus rhinocerotis might promote intestinal mucosal wound healing. AIM OF THE STUDY: This study aimed to investigate the effect and mechanism of ß-glucan prepared from L. rhinocerotis using an enzymatic method on epithelial restitution during intestinal mucosal damage. MATERIALS AND METHODS: Based on FT-IR, MALDI-TOF-MS, HPSEC-MALLS-RID, and AFM, the structure of polysaccharides from L. rhinocerotis was analysed. In addition, polysaccharides were used to test for wound healing activity in IEC-6 cells by measuring cell migration, proliferation, and expression of cell division control protein 42, Rac-1, RhoA, and Par-3. RESULTS: ß-glucan was extracted using enzyme-assisted extraction, and a yield of approximately 8.5 ± 0.8% was obtained from the dried biomass. The ß-glucan extracted by enzyme-assisted extraction (EAE) of polysaccharides was composed entirely of D-glucose with a total carbohydrate content of 95.5 ± 3.2%. The results of HPLC, FTIR, and MALDI-TOF-MS analyses revealed EAEP to be confirmed as ß-glucan. The molecular weight of prepared ß-glucan was found to be 5.315 × 104 g/mol by HPSEC-MALLS-RID. Furthermore, mucosal wound healing studies showed that the treatment of IEC-6 with a ß-glucan concentration of 200 µg/mL promoted cell migration and proliferation, and it enhanced the protein expression of cell division control protein 42, Rac-1, RhoA, and Par-3. CONCLUSIONS: The present study reveals that the prepared ß-glucan accelerates intestinal epithelial cell proliferation and migration via activation of Rho-dependent pathway. Hence, ß-glucan can be employed as a prospective therapeutic agent for the treatment of diseases associated with gastrointestinal mucosal damage, such as peptic ulcers and inflammatory bowel disease.

Polysaccharides from Gracilaria lemaneiformis promote the HaCaT keratinocytes wound healing by polarised and directional cell migration.

In this study, we evaluated the wound healing activity of polysaccharides fractions from Gracilaria lemaneiformis. Three purified fractions, namely, GLP-1, GLP-2 and GLP-3 were obtained from anion-exchange chromatography and were evaluated for their cell proliferation activity. Among the three fractions, the fraction GLP-2 promoted cell proliferation at a concentration of 100 µg/mL. Further, chemical and structural analysis of GLP-2 revealed it to be a homogenous and repeating structure of alternating 4-linked 3,6-anhydro-α-L-galactopyranosyl and 3-linked ß-d-galactopyranosyl units with sulfate residues. Further, we analysed the wound healing activities of the fraction GLP-2 and its underlying mechanisms. The results showed that GLP-2 promotes cell proliferation and migration through activation of PI3 K/aPKC signaling during human keratinocytes wound healing. Based on the findings in this study, we concluded that the wound healing activities of the GLP-2 fraction can provide the scientific basis for the development of G. lemaneiformis polysaccharides based product for wound management.

Ultrasonic/microwave-assisted extraction, simulated digestion, and fermentation in vitro by human intestinal flora of polysaccharides from Porphyra haitanensis.

In this study, we employed a response surface methodology to optimize the ultrasonic/microwave-assisted extraction (UMAE) conditions of Porphyra haitanensis polysaccharides (PHP), and subjected it to a stimulated in vitro digestion and fermentation model in order to investigate the digestion properties of PHP and the effects on human intestinal flora. The optimum extraction conditions consisted of an extraction time of 29.64 min, extraction temperature of 79.94 °C, and solid-liquid ratio of 1:41.79 g/mL. Under these conditions, the maximum yield of PHP predicted was 20.98%. The ζ-potential and thermal properties analysis verified that PHP was a negatively charged polymer, and possessed good thermal stability. Meanwhile, PHP was not digested in vitro by human saliva, simulated gastric and small intestinal juice. Furthermore, PHP modulated the microbiome structure, mainly increasing the relative abundance of Bacteroides and decreasing in the Escherichia_Shigella group. LEfSe analysis illustrated that Bacteroides, Lachnospiraceae_UCG_006 and Bacteroidales_S24_7_group could serve as potential biomarkers for the PHP supplement. This current study proved that the UMAE method was a highly efficient method to extract PHP to the maximum extent, and also provided insight concerning the stability performance of PHP and its prospects for application as a prebiotics candidate in the functional food industries.

Physicochemical characterization and antioxidant activity of sulphated polysaccharides derived from Porphyra haitanensis.

This study was designed to fully characterize Porphyra haitanensis polysaccharides, and to evaluate their antioxidant activity. The polysaccharides primarily contained galactose and 3,6-anhydrogalactose in a molar ratio of 1.2:1.0, respectively and sulfate content about 3.8%. The molecular weight of polysaccharides is 2.5 × 105 Da. Scanning electron microscopy and atomic force microscopy of the polysaccharides pointed towards an irregular network with more or less hexagonal and a few rectangular pores. The chemical structure was confirmed through Fourier transform infrared spectroscopy, and 1D and 2D nuclear magnetic resonance structural characterization wherein â†’ 4-3,6-anhydro-α-L-galactopyranose-(1 â†’ 3)-ß-D-galactopyranose segments. The extracted polysaccharides revealed relatively high 2, 2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical scavenging activity (53.16% at 2 mg/mL), moderate 2,2-diphenyl-1-picrylhydrazyl radical scavenging efficacy (34.63% at 2 mg/mL), and low hydroxyl radical scavenging potential (23.80% at 2 mg/mL). Further purification of these polysaccharides, hence, is advised for their potential role as antioxidants in the food, pharmaceutical and cosmeceutical industry.

Qualitation and quantification of specific polysaccharides from Panax species using GC-MS, saccharide mapping and HPSEC-RID-MALLS.

The objective of this study was to qualify and quantify the specific polysaccharides in Panax spp. The analyses of specific polysaccharides were performed by using GC-MS, saccharide mapping and high performance size exclusion chromatography (HPSEC) coupled with multi angle laser light scattering (MALLS) and refractive index detector (RID). Results showed that compositional monosaccharides were the same in different species of Panax and composed of rhamnose, arabinose, galacturonic acid, mannose, glucose, and galactose. Saccharide mapping results showed that glycosides linkages, which existed in specific polysaccharides from Panax spp., were similar. Additionally, the content of specific polysaccharides of P. ginseng, P. notoginseng and P. quinquefolium were 17.9-20.5mg/g, 11.9-15.0mg/g, and 9.9-13.3mg/g, respectively. P. ginseng, P. notoginseng, and P. quinquefolium could be clustered into three groups using both hierarchical cluster analysis and principal component analysis. The results possessed great potential in characterization and content determination of specific polysaccharides in Panax spp.

Qualitative and quantitative analysis of specific polysaccharides in Dendrobium huoshanense by using saccharide mapping and chromatographic methods.

Qualitative and quantitative analysis of specific polysaccharides from ten batches of Dendrobium huoshanense were performed using high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector (HPSEC-MALLS-RID), gas chromatography-mass spectrometry (GC-MS), nuclear magnetic resonance (NMR) and saccharide mapping based on polysaccharides analysis by using carbohydrate gel electrophoresis (PACE) and high performance thin layer chromatography (HPTLC). Results showed that molecular weights, the radius of gyrations, and contents of specific polysaccharides in D. huoshanense were ranging from 1.16×10(5) to 2.17×10(5)Da, 38.8 to 52.1nm, and 9.9% to 19.9%, respectively. Furthermore, the main monosaccharide compositions were Man and Glc. Indeed, the main glycosidic linkages were ß-1,4-Manp and ß-1,4-Glcp, and substituted with acetyl groups at O-2 and O-3 of 1,4-linked Manp. Moreover, results showed that PACE and HPTLC fingerprints of partial acidic and enzymatic hydrolysates of specific polysaccharides were similar, which are helpful to better understand the specific polysaccharides in D. huoshanense and beneficial to improve their quality control. These approaches could also be routinely used for quality control of polysaccharides in other medicinal plants.

Comparison and Characterization of Compounds with Antioxidant Activity in Lycium barbarum Using High-Performance Thin Layer Chromatography Coupled with DPPH Bioautography and Tandem Mass Spectrometry.

Methanol extracts from 50 batches of Lycium barbarum (L. barbarum, wolfberry) in China were compared and characterized using high-performance thin-layer chromatography coupled with 2,2-diphenyl-1-picrylhydrazyl (DPPH) bioautography (HPTLC-DPPH) and electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI-Q-TOF-MS/MS), respectively. Results showed that similar components occupying the major antioxidant activity existed in L. barbarum collected from different origins. However, the average antioxidant capacities of methanol extracts of L. barbarum collected in Ningxia were significantly higher than those of Qinghai, Xinjiang, Inner Mongolia, and Gansu, which may contribute to rational use of L. barbarum in China. Furthermore, the chemical structure of compound with the highest antioxidant capacity was tentatively identified as 2-O-ß-d-glucopyranosyl-l-ascorbic acid using ESI-Q-TOF-MS/MS analysis, which possessed high potentials to be used as an antioxidant biomarker for the quality control of L. barbarum. Results are helpful for the bioactivity-based quality control of L. barbarum, and beneficial for the improvement of their performance in functional/health foods area, suggesting that HPTLC-DPPH bioautography with ESI-Q-TOF-MS/MS could be used as a routine approach for quality control of antioxidant components in L. barbarum.

A rapid and accurate method for the quantitative estimation of natural polysaccharides and their fractions using high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector.

In this study, a rapid and accurate method for quantitative analysis of natural polysaccharides and their different fractions was developed. Firstly, high performance size exclusion chromatography (HPSEC) was utilized to separate natural polysaccharides. And then the molecular masses of their fractions were determined by multi-angle laser light scattering (MALLS). Finally, quantification of polysaccharides or their fractions was performed based on their response to refractive index detector (RID) and their universal refractive index increment (dn/dc). Accuracy of the developed method for the quantification of individual and mixed polysaccharide standards, including konjac glucomannan, CM-arabinan, xyloglucan, larch arabinogalactan, oat ß-glucan, dextran (410, 270, and 25 kDa), mixed xyloglucan and CM-arabinan, and mixed dextran 270 K and CM-arabinan was determined, and their average recoveries were between 90.6% and 98.3%. The limits of detection (LOD) and quantification (LOQ) were ranging from 10.68 to 20.25 µg/mL, and 42.70 to 68.85 µg/mL, respectively. Comparing to the conventional phenol sulfuric acid assay and HPSEC coupled with evaporative light scattering detection (HPSEC-ELSD) analysis, the developed HPSEC-MALLS-RID method based on universal dn/dc for the quantification of polysaccharides and their fractions is much more simple, rapid, and accurate with no need of individual polysaccharide standard, as well as free of calibration curve. The developed method was also successfully utilized for quantitative analysis of polysaccharides and their different fractions from three medicinal plants of Panax genus, Panax ginseng, Panax notoginseng and Panax quinquefolius. The results suggested that the HPSEC-MALLS-RID method based on universal dn/dc could be used as a routine technique for the quantification of polysaccharides and their fractions in natural resources.

Mycelia extracts of fungal strains isolated from Cordyceps sinensis differently enhance the function of RAW 264.7 macrophages.

ETHNOPHARMACOLOGICAL RELEVANCE: Cordyceps sinensis, an entomogenous fungus used in traditional Chinese medicine with multiple pharmacological activities. However, its usage has been limited due to the high price and short supply. Isolate of fungi strains from natural Cordyceps sinensis to achieve a large-scale production by fermentation is an alternative choice. The aim of this study was to investigate and compare the effects of mycelia extracts of different fungal stains isolated from natural Cordyceps sinensis on macrophage functions in vitro. MATERIALS AND METHODS: Macrophages' proliferation, phagocytosis, nitric oxide (NO) production, cytokines secretion, iNOS, NF-κB p65 activation and translocation were investigated by the MTT assay, flow cytometry assay, Griess reagent method, ELISA, western blot and immunostaining assay, respectively. RESULTS: The results showed that the effects of cultured Cordyceps mycelia of different fungal strains isolated from natural Cordyceps sinensis on macrophages greatly variant. Among 17 Cordyceps aqueous extracts, only five extracts (UM01, QH11, BNQM, GNCC and DCXC) could significantly increase cell proliferation and NO production of RAW 264.7 mouse macrophages. Moreover, the five extracts, especially UM01 and QH11, significantly enhanced phagocytosis and promoted cytokines release of macrophages. Polysaccharides in cultured UM01 mycelia were found to be the main immune stimulating compounds. CONCLUSIONS: The variation of biological effects of fermented mycelia of different fungal strains from natural Cordyceps sinensis may be derived from their chemical diversity, especially polysaccharides, which need further study in future.

Effects of polysaccharides from different species of Dendrobium (Shihu) on macrophage function.

Dendrobium spp. are precious medicinal plants, used in China for thousands of years as health foods and nutrients. Polysaccharides are the main effective ingredients in Dendrobium plants. In this study, the chemical characteristics and the effects of crude polysaccharides (CPs) from five species of Dendrobium on macrophage function were investigated and compared in vitro for the first time. Chemical characteristic studies showed that CPs from different species of Dendrobium were diverse, displaying widely varied Mw distributions and molar ratios of monosaccharides. Their effects on macrophage functions, such as promoting phagocytosis, release of NO and cytokines IL-1α, IL-6, IL-10 and TNF-α, were also different. Moreover, CPs from D. officinale, especially collected from Yunnan Province, exerted the strongest immunomodulatory activities and could be explored as a novel potential functional food. The diverse chemical characteristics of CPs from different species of Dendrobium might contribute to their varied effects on macrophage functions, which should be further investigated.

Chromatography in characterization of polysaccharides from medicinal plants and fungi.

Polysaccharides isolated from medicinal plants and fungi exhibit multiple pharmacological activities. The biological activities of polysaccharides depend on their chemical characteristics. However, characterization of polysaccahrides is a challenge because of their complicated structure and macromolecular mass. In this review, chromatography in characterization of polysaccharides, including physicochemical characterization (purity, molecular mass, and distribution), structural characterization (constituent monosaccharide composition and the ratio, the features of glycosidic linkages), and fingerprint of polysaccharides (acidic and enzymatic hydrolysates), from medicinal plants and fungi were reviewed and discussed according to the publications collected in Web of Science since 2007. The perspective for characterization of polysaccharides has also been described.

Structure and protective effect on UVB-induced keratinocyte damage of fructan from white garlic.

The fructose polymer fructan was extracted from white garlic and fractionated using DEAE cellulose 52 and Sephadex G-100 columns to characterize its chemical composition and protective effect against ultraviolet radiation b (UVB) induced human keratinocyte (HaTaC) damage. Gel permeation chromatography, high performance anion exchange chromatography, infrared spectroscopy and 1D and 2D nuclear magnetic resonance spectroscopy were used to determine the chemical composition and functional characteristics of the garlic fructan (GF). GF was a homogeneous polysaccharide with a molecular weight of 4.54 × 10(3)Da. It was a member of the 1-kestose family, and it was composed of fructose and glucose at a ratio of 14:1. The main chain of GF was composed of (2→1)-ß-D-fructopyranose linked to a terminal (2→1)-α-D-glucopyranose at the non-reducing end and a (2→6)-ß-D-fructopyranose branched chain. The degree of polymerization was 28. Preliminary tests described herein indicated that GF may be effective in protecting HaTaC from UVB-induced damage.