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Two polysaccharide fractions (GLSB50 and GLSB70) with total sugar content of 82.07 wt% and 53.79 wt%, respectively, were obtained from the water extracts of unbroken Ganoderma lucidum spores by sequential ethanol precipitation treatment. Compared with GLSB70, GLSB50 exhibited better activity on stimulation of humoral immune responses in immunosuppressed mice. A novel β-D-glucan (GLSB50A-III-1) with weight average molecular weight (M w ) of 1.93 × 10 5  g/mol was purified from GLSB50 through chromatography separation. The exponent α value of Mark-Houwink-Sakurada equation was calculated to be 0.13, indicating that GLSB50A-III-1 presented globular spheres conformation in aqueous solution. Structural analysis showed that GLSB50A-III-1 mainly consisted of (1 → 3), (1 → 4), (1 → 6)-linked β-d-glucose residues in the backbone, with two single β-D-Glcp attached at O-6 of β-(1 → 3) and β-(1 → 4)-linked residues separately as side chains. The repeat unit of GLSB50A-III-1 was deduced as follows., (Copyright © 2021. Published by Elsevier B.V.)

The polysaccharide is the main active substance contained in Hericium erinaceus and is commonly used in the treatment of neurasthenia, tumors, and digestive diseases. Six intracellular polysaccharide components were obtained from H. erinaceus fruiting bodies cultivated by ARTP (atmospheric and room temperature plasma) mutagenic strain (321) and the original strain (0605), respectively. This study was designed to investigate the physicochemical characteristics of these polysaccharide components and their potential immunomodulatory activities on RAW264.7 macrophages. The results showed that the yield of fruiting body cultivated by mutated strain increased by 22% and the polysaccharide content improved by 16% compared with the original one owing to ARTP mutagenesis. The molecular weight distribution and the monosaccharide compositions of polysaccharide components from H. erinaceus induced by ARTP mutagenesis were significantly different from that of the original one. The NO, IL-6, IL-10, IL-1β, and TNF-α production activities of macrophages were enhanced by stimulation of 20% ethanol precipitated polysaccharides from H. erinaceus induced by ARTP mutagenesis. These results indicated that ARTP is an efficient and practical method for high polysaccharide content breeding of the H. erinaceus strain and this provided a reference for obtaining high quality resources and healthy product development from H. erinaceus ., Competing Interests: The authors declare no conflict of interest.

Ganoderma, named lingzhi in China, has been used for centuries as drug and nutraceutical to treat diseases. Based on our research and other literatures, the chapter summarizes the progress of preparation, structural features and properties, bioactivities of Ganoderma polysaccharides. The aim is to provide a comprehensive source of information for researchers and consumers of Ganoderma, so they can better understand Ganoderma polysaccharides and their biological activities. In addition, more clinical studies should be carried out to meet the criteria for new drug development, and more convincing scientific data should be provided. In addition, on the basis of a large number of studies on Ganoderma polysaccharides, we suggest that more clinical studies should be carried out so that Ganoderma can be better recognized and applied all over the world.

Molecular weight (Mw) distributions of polysaccharides from the fruiting bodies of different Ganoderma lucidum strains and G. sinense were investigated and compared using high-pressure size exclusion chromatography/multiangle laser light scattering/refractive index analysis. Results showed that there were big differences in the Mw distributions and characteristics of polysaccharides from 2 species of Ganoderma. All tested G. lucidum materials exhibited similar polysaccharide distributions and similar characteristics for each fraction. The fraction with highest Mw (peak 1) was identified as β-(1→3)-linked D-glucan with (1→6)-β-D-glucopyranosyl side branches. G. sinense fruiting bodies did not include the β-D-glucan when compared with G. lucidum. A high-pressure size exclusion chromatography method was developed and applied to determine the amount of high-Mw β-D-glucan in G. lucidum fruiting bodies. Results indicated that there was no obvious relationship between β-D-glucan content and the genetic similarity of G. lucidum. The strain labeled "Longzhi no. 2" was determined to possess the largest amount of β-D-glucan: 8.2 mg/mL based on the dry weight of fruiting bodies. The β-D-glucan content in the hot water extract of Longzhi no. 2 reached 17.05%. For the "Hunong no. 1" strain, the β-D-glucan content in log-cultivated fruiting bodies was much higher than that in bag-cultivated ones. This method could be used to improve quality control of polysaccharides in G. lucidum.

HPB-3, a heteropolysaccharide, with a mean molecular weight of 1.5×10(4)Da, was obtained from the maturating-stage IV, V and VI fruiting body of Hericium erinaceus, exhibited higher macrophages stimulation activities, was able to upregulate the functional events mediated by activated macrophages, such as production of nitric oxide (NO). Monosaccharide composition analysis showed that HPB-3 comprised l-fucose, d-galactose and d-glucose in the ratio of 5.2:23.9:1. Its chemical structure was characterized by sugar and methylation analysis, along with (1)H and (13)C NMR spectroscopy, including (1)H-(1)H COSY, TOCSY, NOESY, HMQC and HMBC experiments. The results indicated that HPB-3 contained a-(1/6)-linked galactopyranosyl backbone, partially with a side chain composed of α-l-fucopyranose at the O-2 position. The predicted primary structure of the polysaccharide was established as below., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Nine polysaccharide fractions were obtained from the fruiting bodies, submerged mycelia, and solid state fermented products of Phellinus baumii using different concentrations of ethanol precipitation. The chemical characteristics and in vitro immunological activities of the nine polysaccharide fractions were compared and studied. Results indicated that the fractions precipitated with 50% ethanol had higher yields of polysaccharides and submerged mycelia contributed to high extraction yields of polysaccharides and possessed higher polysaccharide contents. HPSEC-MALLS-RI analysis showed that the molecular weight (Mw) of polysaccharide fractions from these three materials decreased with the increasing of precipitated ethanol concentration. The Mw of fruiting body polysaccharide fractions ranged from 1.98×10(4)Da to 1.89×10(6)Da. Large-molecular-weight polysaccharides (from 2.11×10(6)Da to 2.01×10(7)Da) were found in submerged mycelia. Some lower-molecular-weight polysaccharide components were found in solid fermented products. Different culture methods contributed to significant differences in monosaccharide components and molar ratios. The 50% ethanol precipitated fractions exhibited more complexity on monosaccharide compositions comparing with fractions precipitated with 30% and 70% ethanol. Polysaccharide fractions derived from submerged mycelia exhibited higher macrophages stimulation activities. Submerged culture was found to be a suitable method to prepare active polysaccharides because of its short culture span and reasonable cost., (Copyright © 2015 Elsevier B.V. All rights reserved.)

We studied the effect of the maturation stage on the chemical compositions and macrophage activation activity of polysaccharides from the culinary-medicinal mushroom Hericium erinaceus. Results showed that total polysaccharides increased, whereas protein content decreased with the maturation stage development of fruiting body. Nine polysaccharide fractions, 3 from each of the maturity stages IV (small fungal spine stage), V (mid-fungal spine stage) and VI (mature), were prepared using the gradient ethanol precipitation method. The polysaccharide fraction HP4A isolated from the maturating-stage (stage IV) fruiting body had a significant difference from the fractions HP5A (stage V) and HP6A (stage VI) in the molecular weight distribution and monosaccharide compositions. Immunostimulating tests revealed that the polysaccharide fraction HP6 isolated from the mature stage (stage VI) fruiting body presented higher macrophage activation activity. Our findings provided important information for the harvest and use of H. erinaceus with higher qualities and functional benefits.

A novel polysaccharide (CP2-S) was purified from Cordyceps militaris fruit bodies by hot water extraction, ethanol precipitation, DEAE-Sepharose Fast Flow and Sephacryl S-400 high-resolution chromatography. The polysaccharide had a molecular weight of 5.938 × 10(6) g/mol and was mainly composed of glucose. CP2-S had carbohydrate content estimated to be 100% using the phenol-sulfuric acid method. Immunostimulating experiments in vitro indicated that CP2-S could stimulate nitric oxide production, phagocytosis, respiratory burst activity, and secretion of interleukin-1β and interleukin-2 of macrophages, suggesting that this water-soluble polysaccharide from the fruit body of C. militaris is a natural immunostimulating polysaccharide with potential for further application.

To obtain a low-molecular-weight polysaccharide with immuno-enhancing activity, hot water extract of Ganoderma lucidum fruit bodies was separated by membrane ultrafiltration, anion exchange, and gel filtration chromatography, and the immunological activities of fractions were assessed on the basis of nitric oxide production by RAW 264.7 macrophages. A novel polysaccharide (TB3-2-2) was successfully isolated and purified. TB3-2-2 is a homogeneous polysaccharide, with a relative molecular weight of 5.11 × 103 Da, identified by high-performance liquid chromatography and was composed of galactose and glucose in a molar ratio of 2:3 determined by high-performance anion exchange chromatography. TB3-2-2 had a carbohydrate content of 99%, as measured using the phenol-sulfuric acid method. Proliferation of mouse spleen lymphocytes and the expression level of interleukin-6 was significantly increased by TB3-2-2. Results indicate that the low-molecular-weight polysaccharide with immunological activity in G. lucidum is worthy of further research and development.

A novel acidic heteropolysaccharide (TAPA1) was purified from hot water extracts of Tremella aurantialba fruiting bodies by DEAE-Sepharose Fast Flow and Sephacryl S-500 High-Resolution Chromatography. The heteropolysaccharide had a molecular weight of ca. 1.35x10(6)Da, and a carbohydrate content estimated to be approximately 98.7% by the phenol-sulfuric acid method. It was composed mainly of d-mannose, d-xylose, and d-glucuronic acid in the ratio of ca. 5:4:1, along with trace amounts of d-galacturonic acid and d-glucose. Monosaccharide compositional analysis and GC-MS of methylated derivatives, combined with (1)H and (13)C NMR spectroscopies (including COSY, TOCSY, NOESY, HSQC, and HMBC spectra), revealed TAPA1 to consist of an alpha-(1-->3)-linked mannopyranosyl backbone, partially substituted at position 4 with xylose side chains, and at position 2 with side chains composed of either xylose, mannose, and glucuronic acid or of xylose and mannose. Bioactivity testing showed that TAPA1 stimulated the proliferation of mouse spleen lymphocytes in vitro.

The rapid development of Lentinula edode industry in China has led to the production of large amounts of waste substrates, which has brought enormous pressure to ecological environment. Thus, it is of great significance to study utilization of spent substrate. In this study, complex enzyme combined with high pressure pretreatment were applied to extract polysaccharide from spent substrate of L. edode in order to improve the utilization efficiency of its residue and explore more biological application of the polysaccharides from spent L. edodes substrate. The optimal extraction conditions were as follows: complex enzyme (cellulase, xylanase, β-glucanase) ratio, 3:2:1; total amount of enzyme, 660 U·g-1; liquid/material ratio, 20 mL/g; temperature, 45 ℃; time, 6 h; The yield of purified polysaccharide (LE-UE) was 5.98% and the content was 51.60%. LE-UE could markedly improve inflammatory damage of LPS-induced macrophages RAW264.7 by inhibiting release of NO and secretion of inflammatory cytokines. Animal experiments showed that LE-UE could markedly improve the histopathological damage and inflammation in ulcerative colitis mice by regulating the level of oxidative stress, and reduce the release of inflammatory cytokines by down-regulating mRNA expression and proteins levels. Therefore, LE-UE has anti-inflammatory activity and may be used as a novel anti-inflammatory agent for treatment. This study could provide scientific evidence for the high-value utilization of spent substrates and also provide theoretical basis for the development of functional feed additives. [ABSTRACT FROM AUTHOR]

Polysaccharides with molecular weights ranging from 1.75 × 103 to 1.14 × 104 g/mol were obtained from the fruit bodies of Ganoderma lucidum. The multiple fingerprints and macrophage immunostimulatory activity of these fractions were analyzed as well as the fingerprint–activity relationship. The correlation analysis of molecular weight and immune activity demonstrated that polysaccharides with molecular weights of 4.27 × 103~5.27 × 103 and 1 × 104~1.14 × 104 g/mol were the main active fractions. Moreover, the results showed that galactose, mannose, and glucuronic acid were positively related to immunostimulatory activity. Additionally, partial least-squares regression and grey correlation degree analyses indicated that three peaks (P2, P3, P8) in the oligosaccharide fragment fingerprint significantly affected the immune activity of the polysaccharides. Hence, these ingredients associated with activity could be considered as markers to assess Ganoderma lucidum polysaccharides and their related products, and the study also provides a reference for research on the spectrum–effect relationship of polysaccharides in the future. [ABSTRACT FROM AUTHOR]

The cell wall polysaccharides were extracted from Sparassis latifolia fruit bodies by acid–alkali and superfine-grinding assisted methods, and the chemical characterization and in vitro immunity activities of these polysaccharide fractions were studied and compared. Results showed that superfine-grinding assisted extraction exhibited the highest yield of polysaccharides (SP, 20.80%) and low β-glucan content (19.35%) compared with alkaline extracts. The results revealed that the 20% ethanol precipitated fraction (20E) from SP was mainly composed of β-(1→3)-glucan and α-(1→4)-glucan. With the increase of ethanol precipitation, the fractions (30E, 40E, 50E) were identified as α-(1→4)-glucan with different molecular weights and conformations. Cell wall polysaccharides extracted through NaOH (NSP) and KOH (KSP) extraction had similar yields with 8.90% and 8.83%, respectively. Structural analysis indicated that the purified fraction from KSP (KSP-30E) was a β-(1→3)-glucan backbone branched with β-(1→6)-Glcp, while the purified fraction from NSP (NSP-30E) mainly contained β-(1→3)-glucan with a small number of α-linked-Glcp. The two fractions both exhibited rigid chain conformation in aqueous solutions. All polysaccharide fractions exerted the activity of activating Dectin-1 receptor in vitro, and the KSP-30E mainly identified as β-(1→3)-glucan with the terminal group via 1→6-linkage attached at every third residue exhibited a stronger enhancing effect than other fractions. Results suggested that KOH extraction could be efficient for the preparation of bioactive β-(1→3, 1→6)-glucan as a food ingredient. [ABSTRACT FROM AUTHOR]

Antibiotic resistance genes (ARGs) are an emerging concern in wastewater treatment plants (WWTPs), as dissemination of ARGs can pose a serious risk to human health. Few studies, however, have quantified ARGs in membrane bioreactors (MBRs), although MBRs have been widely used for both municipal and industrial wastewater treatment. To reveal the capacity of MBRs for removal of ARGs and the response of membrane fouling after antibiotic exposure, five typical ARG subtypes ( sul I, sul II, tet C, tet X and ere A) and int 1 were quantified affiliated by systematic membrane foulants analysis in a laboratory-scale anoxic/aerobic membrane bioreactor (A/O-MBR). Sulfamethoxazole and tetracycline hydrochloride additions increased ARG abundances by 0.5–1.4 orders of magnitude in the activated sludge, while the ARG removal performance of the membrane module remained stable (or even increased with ARG absolute abundance in several cases), with the abundance of removed ARGs ranging from 0.6 to 5.6 orders of magnitude. Specifically, the distribution of ARGs in membrane foulants accounted for 13%–25% of the total absolute abundance of all tested MBR samples. Indeed, substantial fouling occurred after the antibiotic additions, with the mean concentrations of soluble microbial product (SMP) and extracellular polymeric substance (EPS) increasing by 340% and 220%, respectively, in a membrane fouling cycle; moreover, the contents of EPS and SMP in the membrane foulants were significantly correlated with the ARG absolute abundance of membrane foulants ( p < 0.05), among which more significant correlations occurred between both the protein and polysaccharide of foulants than that with humic acid. The dense membrane fouling layer and the membrane itself constituted dual barriers that effectively avoided the leakage of ARGs from the membrane module. Our findings provide fundamental insights into the proliferation and removal of ARGs in MBR systems, and highlight the contribution of membrane fouling to ARG removals in terms of the potential of MBR as an effective strategy to reduce ARG levels in WWTP effluent. [ABSTRACT FROM AUTHOR]

A proteoglycan LEPS1 was firstly isolated and purified from the spent substrate of Lentinula edodes , an agricultural waste that may cause environmental pollution. The average molecular weight of LEPS1 was 1.18 × 104 g/mol, and carbohydrate moiety (88.9 %) was composed of glucose, arabinose, galactose, xylose and mannose at a molar ratio of 1.2:1.2:1.0:2.3:1.1. The protein moiety (8.5 %) of LEPS1 was bonded to the polysaccharide chain via O-glycosidic linkage. LEPS1 could significantly improve the inflammatory injury of LPS stimulated RAW264.7 macrophages by inhibiting the secretion of NO and decreasing the levels of pro-inflammatory factors (TNF-α, IL-1β and IL-6). LEPS1 inhibited JAK-STAT1 and p38 MAPK signaling pathway via modulating JAK expression, phosphorylation of STAT1 and phosphorylation of p38, respectively. Moreover, LEPS1 could promote the expression of CD 206 and IL-10 which were the markers for repairing macrophages. Overall, LEPS1 had anti-inflammatory activity and can potentially treat as a novel anti-inflammation agent. This work could provide scientific basis and valuable information for the highly efficient utilization of spent L. edodes substrates as the by-product in mushroom industries. [Display omitted] • A proteoglycan LEPS1 was obtained from spent substrate of Lentinula edodes. • The protein moiety of LEPS1 was bonded to polysaccharide chain via O-glycosidic linkage. • The average molecular weight of LEPS1 was 1.18 × 104 g/mol. • LEPS1 could improve the inflammatory injury of LPS stimulated macrophages. • Anti-inflammatory activity of LEPS1 was mediated by p38-MAPK and JAK-STAT1 pathways. [ABSTRACT FROM AUTHOR]

Abstract: Three water-soluble polysaccharide fractions (IOP40, IOP60 and IOP80) were isolated by using different concentrations of alcohol precipitation from Inonotus obliquus sclerotia. Their physicochemical properties, including total sugar content, protein content, monosaccharide composition and percentage were analyzed. And their in vitro antioxidant capacities were investigated in terms of reducing power assay and scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals, hydroxyl radicals, superoxide anion radicals and hydrogen peroxide (H2O2). In general, three polysaccharide fractions exhibited increasing antioxidant activity with increasing concentration at the ranges of tested dosage. The orders of reducing power, DPPH-scavenging capacity, H2O2-scavenging capacity, and hydroxyl-scavenging activity were all IOP60>IOP40>IOP80. These findings demonstrated that three polysaccharide fractions extracted from I. obliquus, especially IOP60, could be employed as natural ingredients in functional food and pharmaceutical industry to alleviate the oxidative stress. [Copyright &y& Elsevier]

Highlights • An acidic heteropolysaccharide (NSKP) was purified from Nostoc sphaeroids kütz. • The complex structure and conformational characteristics of NSKP was elucidated. • The immunological activity of GLSWA-I was tested in vitro. Abstract A water soluble Nostoc sphaeroids kütz polysaccharide (NSKP) was extracted and purified by precipitating hot-water extract from Nostoc sphaeroids kütz with 40% (v/v) ethanol. The yield was 40.10% (w/w), and weight average molecular weight (M w) was determined as 1.31 × 105 g/mol. Cell culture study showed that NSKP could stimulate the NO release from RAW264.7 macrophages. The ρ value (R g / R h) of NSKP was calculated as 1.50, indicating flexible random coils conformation of NSKP in aqueous solution. According to methylation analysis, and 1D & 2D NMR spectra, the molecular structure of polysaccharide NSKP was deduced as below: [ABSTRACT FROM AUTHOR]