In order to provide a foundation for the development and application of Ophiocordyceps gracilis and increase the new resources of cordyceps,an asexual Paraisaria dubia was isolated from an O. gracilis fruit body. After 10 days of liquid fermentation,white globular mycelium and clear transparent fermentation were produced. The mycelium was extracted by hot water and precipitated with ethanol to obtain intracellular crude polysaccharide. The protein was deproteinized to obtain deproteinized polysaccharide. The intracellular pure polysaccharide was purified by Sepharose 4 B column chromatography and were analyzed by UV,IR,1 H-NMR,and13 CNMR data,as well as GC and HPLC. The results showed that the intracellular polysaccharide of P. dubia was composed of glucose,galactose and mannose with a molar ratio of 25. 54 ∶2 ∶1. It was a β-configuration glycosylic bond,containing pyranoside. The initial connection of polysaccharide was β(1→2)(1→4)(1→6) connection. This experiment provides a theoretical basis for the development and application of P. dubia.
Fungal Polysaccharides ; chemistry ; Galactose ; Glucose ; Hypocreales ; chemistry ; Mannose ; Mycelium ; chemistry

Metabolic syndrome characterized by obesity, hyperglycemia and liver steatosis is becoming prevalent all over the world. Herein, a water insoluble polysaccharide (WIP) was isolated and identified from the sclerotium of Poria cocos, a widely used Traditional Chinese Medicine. WIP was confirmed to be a (1-3)-β-D-glucan with an average Mw of 4.486 × 10 Da by NMR and SEC-RI-MALLS analyses. Furthermore, oral treatment with WIP from P. cocos significantly improved glucose and lipid metabolism and alleviated hepatic steatosis in ob/ob mice. 16S DNA sequencing analysis of cecum content from WIP-treated mice indicated the increase of butyrate-producing bacteria Lachnospiracea, Clostridium. It was also observed that WIP treatment elevated the level of butyrate in gut, improved the gut mucosal integrity and activated the intestinal PPAR-γ pathway. Fecal transplantation experiments definitely confirmed the causative role of gut microbiota in mediating the benefits of WIP. It is the first report that the water insoluble polysaccharide from the sclerotium of P. cocos modulates gut microbiota to improve hyperglycemia and hyperlipidemia. Thereby, WIP from P. cocos, as a prebiotic, has the potential for the prevention or cure of metabolic diseases and may elucidate new mechanism for the efficacies of this traditional herbal medicine on the regulation of lipid and glucose metabolism.
Animals ; Bacteria ; classification ; genetics ; isolation & purification ; metabolism ; Butyrates ; metabolism ; Fatty Liver ; drug therapy ; Fungal Polysaccharides ; chemistry ; pharmacology ; therapeutic use ; Gastrointestinal Microbiome ; drug effects ; genetics ; Hyperglycemia ; drug therapy ; Hyperlipidemias ; drug therapy ; Intestines ; drug effects ; microbiology ; Male ; Metabolic Syndrome ; drug therapy ; Mice ; Mice, Obese ; Prebiotics ; Wolfiporia ; chemistry

Near infrared spectroscopy combined with chemometrics methods was used to distinguish Ganoderma lucidum samples collected from different origins, and a prediction model was established for rapid determine polysaccharides contents in these samples. The classification accuracy for training dataset was 96.87%, while for independent dataset was 93.33%; as for the prediction model, 5-fold cross-validation was used to optimize the parameters, and different signal processing methods were also optimized to improve the prediction ability of the model. The best square of correlation coefficients for training dataset was 0.965 4, and 0.851 6 for validation dataset; while the root-mean-square deviation values for training dataset and validation dataset were 0.018 5 and 0.023 6, respectively. These results showed that combining near infrared spectroscopy with suitable chemometrics approaches could accuracy distinguish different origins of G. lucidum samples; the established prediction model could precious predict polysaccharides contents, the proposed method can help determine the activity compounds and quality evaluation of G. lucidum.
Fungal Polysaccharides ; analysis ; Geography ; Least-Squares Analysis ; Reishi ; chemistry ; Spectroscopy, Near-Infrared

OBJECTIVEThe present study is concerning qualitative and quantitative detection of Poria cocos quality based on FT-near infrared (FT-NIR) spectroscopy combined with chemometrics.

METHODThe Poria cocos polysaccharides contents were determined by UV. Transmission mode was used in the collection of NIR spectral samples. The pretreatment method was first derivation and vector normalization. Then principal component analysis (PCA) was used to build classification model and partial least square (PLS) to build the calibration model.

RESULTThe results showed that conventional criteria such as the R, root mean square error of calibration (RMSEC), and the root mean square error of prediction (RMSEP) are 0.944 0, 0.072 1 and 0.076 2, respectively. The misclassified sample is 0 using the qualitative model built by PCA.

CONCLUSIONThe prediction models based on NIR have a better performance with high precision, good stability and adaptability and can be used to predict the polysaccharose content of Poria cocos rapidly, which can provide a fast approach to discriminate the different parts of Poria cocos.

Fungal Polysaccharides ; analysis ; Least-Squares Analysis ; Poria ; chemistry ; Principal Component Analysis ; Spectroscopy, Near-Infrared ; methods

Due to substantial morbidity and high complications, diabetes mellitus is considered as the third "killer" in the world. Medicinal fungal polysaccharides, as water-soluble macromolecular substances with low toxicity, exhibit diversified pharmacological actions such as immune regulation, anti-tumor, antivirus, antioxidant, anti-aging, hypoglycemic effect and improving liver and kidney function. In recent year, a number of investigators reported medicinal fungal polysaccharides showed good anti-diabetes and hypoglycemic activity, and their acting mechanisms involved in glycometabolism and β cell function, e. g. promoting glycogen synthesis, promoting glycolysis, inhibiting the activity of α-glucosidase, promoting insulin secretion, increasing insulin sensitivity, enhancing antioxidation. Therefore, the hypoglycemic activity and its mechanisms of action of medicinal fungal polysaccharides showed characteristics of multiple effects, multi-target, and multi-pathway regulation. These finding suggest that medicinal fungal polysaccharides are a promising source for the development of discovery of anti-diabetic agent.
Animals ; Carbohydrate Metabolism ; drug effects ; Fungal Polysaccharides ; pharmacology ; Humans ; Hypoglycemic Agents ; pharmacology ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; Oxidative Stress ; drug effects

Aims: Polysaccharide of Pleurotus ostreatus is one of the fungal polysaccharide which has been widely studied, produced by extracting the fruiting body. An alternative method for producing polysaccharide of P. ostreatus directly from the mycelia instead of the fruiting body is through submerged culture. This study was aimed to determine the optimum submerged culture conditions for producing biomass and intracellular polysaccharide of the oyster mushroom. Methodology and results: P. ostreatus BPPTCC 6017 was collected from traditional mushroom farm in West Java, Indonesia. Submerged fermentation was conducted in 1000 mL medium (2 L flask). Four variables were tested: temperature, pH, agitation, and fermentation time, using central composite design of the response surface methodology. Mycelial biomass produced, was extracted to obtain water-soluble and alkali-soluble polysaccharide. Experimental data obtained were fitted to a second-order polynomial equation using multiple regression analysis and also analysed by appropriate statistical methods. The 3-D response surface plots derived from the mathematical models were applied to determine the optimum conditions: temperature 27.89 °C, initial pH medium 5.49, agitation 124.08 rpm, and fermentation time 11.44 days. The predicted results of the models were 33.75 g/L mycelia, 0.33 g/L water-soluble polysaccharide, and 0.64 g/L alkali-soluble polysaccharide. Those results were then verified on the optimum conditions, and produced 32.00±1.25 g/L mycelia, 0.29±0.01 g/L water-soluble polysaccharide and 0.60±0.02 g/L alkali-soluble polysaccharide, were close to the theoretical predictions. Conclusion, significance and impact study: The present study was a first effort to assess and obtain the optimum conditions for producing the biomass and polysaccharides of the strain P. ostreatus BPPTCC 6017 using submerged fermentation
Fungal Polysaccharides

A novel Pleurotus nebrodensis polysaccharide (PN-S) was purified and characterized, and its immune-stimulating activity was evaluated in RAW264.7 macrophages. PN-S induced the proliferation of RAW264.7 cells in a dose-dependent manner, as determined by the MTT assay. After exposure to PN-S, the phagocytosis of the macrophages was significantly improved, with remarkable changes in morphology being observed. Flow cytometric analysis demonstrated that PN-S promoted RAW264.7 cells to progress through S and G2/M phases. PN-S treatment enhanced the productions of interleukin-6 (IL-6), nitric oxide (NO), interferon gamma (INF-γ), and tumor necrosis factor-α (TNF-α) in the macrophages, with up-regulation of mRNA expressions of interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), interferon gamma(INF-γ) and tumor necrosis factor-α (TNF-α) being observed in a dose-dependent manner, as measured by qRT-PCR. In conclusion, these results suggest that the purified PN-S can improve immunity by activating macrophages.
Animals ; Cell Cycle ; immunology ; Cell Line ; Cell Proliferation ; drug effects ; Fungal Polysaccharides ; pharmacology ; Immunity ; drug effects ; Interferon-gamma ; biosynthesis ; metabolism ; Interleukin-6 ; biosynthesis ; metabolism ; Macrophages ; immunology ; metabolism ; Mice ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; metabolism ; Pleurotus ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; biosynthesis ; metabolism ; Up-Regulation

This study was conducted to investigate the physicochemical properties of Polyporus umbellatus sclerotial exudate. Morphological characteristics of the sclerotia and its exudate were observed during different stages of sclerotial formation. The pH of the exudate was detected at different time during cultivation. A phenol-sulfuric acid method was employed to determine the polysaccharide content of P. umbellatus sclerotial exudate during cultivating time. Additionally, the protein content was measured by means of BCA protein assay. Furthermore, CAT content was detected using ultraviolet absorption method. That the protein content of the exudate and CAT specific activity rose gradually during the passage of the cultivating time indicated a high level of oxidative stress during P. umbellatus sclerotial exudate formation. The results showed that the pH of the exudate increased gradually and then dropped down during sclerotial formation. That the pH of the exudate maintained the acidity state during the cultivation indirectly indicated that acidic environment would help sclerotial formation. The exudate produced gradually and was absorbed by the sclerotia itself.
Culture Media ; chemistry ; metabolism ; Fungal Proteins ; chemistry ; metabolism ; Fungi ; chemistry ; metabolism ; Hydrogen-Ion Concentration ; Medicine, Chinese Traditional ; methods ; Oxidative Stress ; Polyporus ; chemistry ; metabolism ; Polysaccharides ; chemistry ; metabolism

OBJECTIVETo reveal the quality variation of polysaccharides, triterpenoids and proteins in spores and fruiting bodies of Ganoderma lucidum from producing areas, different varieties, harvesting parts and periods, and wall-breaking treatments.

METHODSpores and fruiting bodies from varieties of Longzhi No. 1 and Hunong No. 1 were collected as test samples, together with wall-broken spores sold in domestic main producing areas. The anthrone-sulfuric acid colorimetric method was used to determine the content of total polysaccharides. The vanillin-glacial acetic acid-perchloric acid colorimetric method was used to determine the content of total triterpenoids. The Lowry method was used to determine the content of total proteins.

RESULTThe content ranges of total polysaccharides, total triterpenoids, and total proteins from 6 domestic main producing areas were 0.40% - 2.25%, 1.36%-3.15% and 0.74% -1.91% respectively. The content ranges of total polysaccharides, triterpenoids, and proteins in the fruiting bodies from 2 varieties cultured in Zhejiang were 0.25% -1.42%, 0.44% -1.42% and 1.82% -3.67% respectively. In addition, the ranges of samples from wall-unbroken spores were 0.41% - 0.91%, 0.09% - 0.12%, 0.78% - 0.90% respectively and wall-broken spores are 1.03% - 2.25%, 1.89% - 3.15%, 0.96% - 1.04% respectively.

CONCLUSIONThere are significant differences in the contents of main chemical ingredients of wall-broken G. lucidum spores saled in the markets. The samples from Zhejiang contain high content of total polysaccharides and triterpenoids, and samples from Fujian contains more proteins. Between the 2 major varieties cultured in Zhejiang, Longzhi No. 1 contains higher content of triterpenoids, but Hunong No. 1 has more polysaccharides. Contents of triterpenoids and polysaccharides from wall-broken spores are much higher than those of fruiting bodies. The stipes from fruiting bodies contains more polysaccharides than those of the pileus, while the triterpenoids contents are higher in the pileus than stipes. The pileus and stipes collected in the second year contain higher content of polysaccharides than the first year's samples, but the contents of triterpenoids are lower. Wall-breaking treatment would significantly improve the extraction and dissolution rate of total triterpenoids and polysaccharides.

Fungal Proteins ; analysis ; Polysaccharides ; analysis ; Reishi ; chemistry ; Spores, Fungal ; chemistry ; Triterpenes ; analysis

To explore one-step method for the preparation of Ganoderma lucidum multicomponent microemulsion, according to the dissolution characteristics of triterpenes and polysaccharides in Ganoderma lucidum, formulation of the microemulsion was optimized. The optimal blank microemulsion was used as a solvent to sonicate the Ganoderma lucidum powder to prepare the multicomponent microemulsion, besides, its physicochemical properties were compared with the microemulsion made by conventional method. The results showed that the multicomponent microemulsion was characterized as (43.32 +/- 6.82) nm in size, 0.173 +/- 0.025 in polydispersity index (PDI) and -(3.98 +/- 0.82) mV in zeta potential. The contents of Ganoderma lucidum triterpenes and polysaccharides were (5.95 +/- 0.32) and (7.58 +/- 0.44) mg x mL(-1), respectively. Sonicating Ganoderma lucidum powder by blank microemulsion could prepare the multicomponent microemulsion. Compared with the conventional method, this method is simple and low cost, which is suitable for industrial production.
Emulsions ; Fungal Polysaccharides ; analysis ; Materia Medica ; chemistry ; Medicine, Chinese Traditional ; Particle Size ; Powders ; Reishi ; chemistry ; Solubility ; Technology, Pharmaceutical ; methods ; Triterpenes ; analysis