Effects of Oleic Acid Addition Methods on the Metabolic Flux Distribution of Ganoderic Acids R, S and T's Biosynthesis.

The effects of oleic acid addition methods on the metabolic flux distribution of ganoderic acids R, S and T's biosynthesis from Ganoderma lucidum were investigated. The results showed that adding filter-sterilized oleic acid in the process of submerged fermentation and static culture is of benefit to the synthesis of ganoderic acids R, S and T. The metabolic fluxes were increased by 97.48%, 78.42% and 43.39%, respectively. The content of ganoderic acids R, S and T were 3.11 times, 5.19 times and 1.44 times higher, respectively, than they were in the control group, which was without additional oleic acid. Ganoderic acids R, S and T's synthesis pathways (GAP), tricarboxylic acid cycles (TCA), pentose phosphate pathways (PP) and glycolysis pathways (EMP) were all enhanced in the process. Therefore, additional oleic acid can strengthen the overall metabolic flux distribution of G. lucidum in a submerged fermentation-static culture and it can reduce the accumulation of the by-product mycosterol. This study has laid an important foundation for improving the production of triterpenes in the submerged fermentation of G. lucidum.

Cordycepin inhibits pancreatic cancer cell growth in vitro and in vivo via targeting FGFR2 and blocking ERK signaling.

Cordycepin (3'-deoxyadenosine) from Cordyceps militaris has been reported to have anti-tumor effects. However, the molecular target and mechanism underlying cordycepin impeding pancreatic cancer cell growth in vitro and in vivo remain vague. In this study, we reported functional target molecule of cordycepin which inhibited pancreatic cancer cells growth in vitro and in vivo. Cordycepin was confirmed to induce apoptosis by activating caspase-3, caspase-9 and cytochrome c. Further studies suggested that MAPK pathway was blocked by cordycepin via inhibiting the expression of Ras and the phosphorylation of Erk. Moreover, cordycepin caused S-phase arrest and DNA damage associated with activating Chk2 (checkpoint kinase 2) pathway and downregulating cyclin A2 and CDK2 phosphorylation. Very interestingly, we showed that cordycepin could bind to FGFR2 (KD = 7.77 × 10-9) very potently to inhibit pancreatic cancer cells growth by blocking Ras/ErK pathway. These results suggest that cordycepin could potentially be a leading compound which targeted FGFR2 to inhibit pancreatic cells growth by inducing cell apoptosis and causing cell cycle arrest via blocking FGFR/Ras/ERK signaling for anti-pancreatic cancer new drug development.

Optimization of Ganoderma lucidum Polysaccharides Fermentation Process for Large-Scale Production.

The objective of this study was to increase the intracellular polysaccharide yield of Ganoderma lucidum. The accordingly optimized fermentation medium by central composite design method contains glucose 40 g L-1, yeast powder 12 g L-1, potassium dihydrogen phosphate 3 g L-1, initial pH 5.5, and inoculum size 10 mL 100 mL-1. Under this condition, the predicted value of intracellular polysaccharide yield was 2.03 g L-1. Shake flask experiments confirmed that the average intracellular polysaccharide yield was 1.98 g L-1 similar to the predicted value. The yields of intracellular polysaccharides in the 5-L and 50-L fermentors were 2.59 g L-1 and 2.65 g L-1, respectively. The molecular weight distribution of intracellular and extracellular polysaccharides obtained was determined by HPSEC-MALLS-RI. The results showed that the weight-average molecular weight of component 1 in the intracellular crude polysaccharide was 4.695 × 106 Da and the mass fraction was 58%. The weight-average molecular weight of component 2 in the extracellular polysaccharide was 5.554 × 104 Da. The mass fraction was 94.9%. The liquid submerged fermentation process of G. lucidum mycelium obtained from this study has effectively increased the yield of intracellular polysaccharides. Its intracellular and extracellular polysaccharides have good immunological activity. Conceivably, the optimized process can be applied for the large-scale production.

Chemical Compounds and Antioxidant Activity of Volatile Oil from the White Jelly Mushroom, Tremella fuciformis (Tremellomycetes).

To fully analyze the composition of volatile oil extracted from Tremella fuciformis, hydrodistillation (HD) and solid phase microextraction (SPME) were adopted simultaneously. In both cases, the analysis was carried out using gas chromatography-mass spectrometry and the antioxidant activity of the volatile oil was determined by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method with rutin as a positive control. Nineteen components in HD and 68 components in SPME were identified, respectively. Moreover, the oil obtained from T. fuciformis by HD indicated that aromatic compounds were a major class (93.5%), followed by the terpenes (5.7%), alkanes (0.4%), and alcohols (0.3%). Among them, butylated hydroxytoluene was the highest concentration (92.5%) of the compounds. The compounds detected by SPME were different from those of HD, and the substances with the largest content were esters (57.7%), followed by alcohols (19.0%), acids (7.0%), and aldehydes (6.3%). Only three of the same substances were detected in both of them, namely borneol, (-)-α-terpineol, and acetic acid. In the DPPH assays, strong antioxidant activity (IC50 = 0.176 mg/mL) was evident in volatile oil from T. fuciformis. Antioxidant activity was positively correlated with the concentration of volatile oil.

[Preliminary Investigation of the Amount, the Molecular Weight and the Activity of Polysaccharides from Chaenomeles Speciosa Fruits in Ethanol Fractional Precipitation].

Chaenomeles speciosa fruits were extracted using water. The extracts were precipitated with 20%~95% (φ) ethanol, respectively. The amount of total polysaccharide was measured with phenol-sulfuric acid method. A method using high-performance size-exclusion chromatography (HPSEC) equipped with multiangle laser-light-scattering photometry (MALLS) and differential refractometry (RI) was presented for determining the molecular weight and molecular weigh distribution. RAW264.7 macrophage were cultured and stimulated with the polysaccharides in vitro and the production of nitric oxide in the cells was determined by the Griess assay. The aim of the study is to determine the amount and the molecular weight of the polysaccharides from Chaenomeles speciosa fruits, and preliminary investigate the immunomodulatory activity, The study provided the basis datas for the further research of Chaenomeles speciosa fruits. , and provided a simple and system method for the research of natural polysaccharide. The ethanol fractional precipitation showed that the order of total polysaccharide content was 95%>80%>40% ≥60%>20%. The results indicated that most polysaccharide from Chaenomeles speciosa fruits might be precipitated when ethanol concentration was up to 95% (T) and the crude polysaccharide purity had risen from 35. 1% to 45. 0% when the concentration of ethanol increased from 20% to 95%. HPSEC-MALLS-RI system showed that all the polysaccharide samples had the similar compositions. They appeared three chromatographic peaks and the retention time were not apparently different. The Mw were 6. 570 X 10(4) g . mol-1 and 1. 393 X 10(4) g . mol-1 respectively, and one less than 10 000 which was failure to obtain accurate values. The molecular weight of the first two polysaccharide distribution index(Mw/Mn)were 1. 336 and 1. 639 respectively. The polysaccharide samples had not exhibited immunomodulatory activity assessed on the basis of nitric oxide production by RAW264. 7 macrophage cells in the experiment.

Nutritional Composition of Three Domesticated Culinary-Medicinal Mushrooms: Oudemansiella sudmusida, Lentinus squarrosulus, and Tremella aurantialba.

The nutritional composition of three recently domesticated culinary-medicinal mushroom species (Oudemansiella sudmusida, Lentinus squarrosulus, and Tremella aurantialba) was evaluated for contents of protein, fiber, fat, total sugar content, amino acid, carbohydrate, and nucleotide components. The data indicated that fruiting bodies of these three mushroom species contained abundant nutritional substances. The protein contents of L. squarrosulus and O. submucida were 26.32% and 14.70%, which could be comparable to other commercially cultivated species. T. aurantialba contained 74.11% of carbohydrate, of which soluble polysaccharide was 40.55%. Oudemansiella sudmusida contained 15.95% of arabitol as the highest sugar alcohol in three mushrooms. These mushrooms also possessed distinct taste by their flavor component composition. Among them, L. squarrosulus contained 10.68% and 9.25% of monosodium glutamate-like and sweet amino acids, which were higher than the other two mushrooms. However, the nucleotide amounts of the three mushrooms were all lower than those of other commercially cultivated mushrooms. Among them, L. squarrosulus contained the highest amount of flavor nucleotides, which was 1.01‰. Results revealed that these three mushroom species are potentially suitable resources for commercial cultivation and healthy food.

Purification, chemical modification and immunostimulating activity of polysaccharides from Tremella aurantialba fruit bodies.

Ultrafiltration and a series of chromatographic steps were used to isolate and purify polysaccharides from Tremella aurantialba fruit bodies. Three crude fractions (TAP50w, TAP10-50w, and TAP1-10w), five semi-purified fractions (TAPA-TAPE), and one purified fraction (TAPA1) were obtained. A sulfated derivative of TAPA1 (TAPA1-s) was prepared by chemical modification. The immunostimulating activity of the polysaccharide fractions in vitro was determined using the mouse spleen lymphocyte proliferation assay. Of the three crude fractions tested, cell proliferation rates were increased most by TAP50w. Furthermore, TAPA1-s was markedly more stimulatory than TAPA1, indicating that sulfonation was an effective way to enhance the immunostimulating activity of polysaccharide.

Structural elucidation of a novel fucogalactan that contains 3-O-methyl rhamnose isolated from the fruiting bodies of the fungus, Hericium erinaceus.

A new heteropolysaccharide, HEPF3, was isolated from the fruiting bodies of Hericium erinaceus. HEPF3 has a molecular weight of 1.9 x 10(4) Da and is composed of fucose and galactose in a ratio of 1:4.12. Compositional analysis, methylation analysis, together with 1H and 13C NMR spectroscopy established that HEPF3 consists of a branched pentasaccharide repeating unit with the following structure: [structure: see text]. HEPF3 also contains a minor proportion of 3-O-methylrhamnose that is thought to terminate the polymer main chain.

[Activation of mouse macrophages by the alkali-extracted polysaccharide from spore of Ganoderma lucidum].

AIM: To investigate the activation of mouse macrophages by the alkali-extracted polysaccharides from the spore of Ganoderma lucidum (LZSBS). METHODS: The mouse macrophages cultured in-vitro were stimulated by LZSBS. IL-1beta and TNF-alpha in the culture supernatants were detected by ELISA. NO production was detected by Griess assay. The percentage of phagocytosis of latex beads by mouse macrophages was counted under microscope. RESULTS: The mouse macrophages stimulated by LZSBS increased in volume and darkened in appearance under phase-contrast microscope. LZSBS-activated mouse macrophages secreted IL-1beta and TNF-alpha produced a large amount of NO. The percentage of phagocytosis of latex beads by mouse macrophages was also significantly increased in the presence of LZSBS. CONCLUSION: LZSBS can activate markedly the mouse macrophages.