Sulphureus Var Research Articles

Acetyl Eburicoic Acid from Laetiporus sulphureus var. miniatus Suppresses Inflammation in Murine Macrophage RAW 264.7 Cells

The basidiomycete Laetiporus sulphureus var. miniatus belongs to the Aphyllophorales, Polyporaceae, and grows on the needleleaf tree. The fruiting bodies of Laetiporus species are known to produce N-methylated tyramine derivatives, polysaccharides, and various lanostane triterpenoids. As part of our ongoing effort to discover biologically active compounds from wood-rotting fungi, an anti-inflammatory triterpene, LSM-H7, has been isolated from the fruiting body of L. sulphureus var. miniatus and identified as acetyl eburicoic acid. LSM-H7 dose-dependently inhibited the NO production in RAW 264.7 cells without any cytotoxicity at the tested concentrations. Furthermore it suppressed the production of proinflammatory cytokines, mainly inducible nitric oxide synthase, cyclooxygenase-2, interleukin (IL)-1β, IL-6 and tumor necrosis factor α, when compared with glyceraldehyde 3-phosphate dehydrogenase. These data suggest that LSM-H7 is a crucial component for the antiinflammatory activity of L. sulphureus var. miniatus.

Four lanostane-type triterpenes from the fruiting bodies of mushroom Laetiporus sulphureus var. miniatus

Two new 3,4-seco-lanostane-type triterpenes, named as 15α-hydroxy-3,4-seco-lanosta-4(28),8,24-triene-3,21-dioic acid (1), 5α-hydroxy-3,4-seco-lanosta-4(28),8,24-triene-3,21-dioic acid 3-methyl ester (2), and one new lanostane triterpene 15α-acetoxylhydroxytrametenolic acid (3) together with a known one versisponic acid D (4) were isolated from the fruiting bodies of Laetiporus sulphureus var. miniatus. Their structures were determined on the basis of extensive spectroscopic methods and comparison with reported data. All four compounds were evaluated for their cytotoxicities against five human cancer cell lines; however, none exhibited inhibitory effects.

Mushrooms Collected from Deogyu Mountain, Muju, Korea and Their Antioxidant Activity

Mushrooms collected from Deogyu mountain, Korea, in 2011, were identified as four classes, four orders, 13 families, 22 genera, and 33 species. In particular, agaricales was most abundant and comprised more than 70%. Their antioxidant activities were estimated using three different bioassay methods, the 2,2′ -azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) radical scavenging assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, and reducing power assay. As a result, the methanol extracts of Stereum ostrea, Laetiporus sulphureus var. miniatus, and Tyromyces sambuceus exhibited potent antioxidant activity in all bioassays tested.

목재부후균에 의한 trans,trans-farnesol의 생물변환

In this study, we screened and evaluated possibility of wood rot fungi as biocatalyst for biotransformation of sesquiterpenes. Screening were performed to select the most promising microorganisms with ability to biotransformation the substrate trans,trans-fanesol. Trans,trans-farnesol which is synthesized precursor of sesquiterpenes was used for resistance test on 19 of wood rot fungi. From the 19 tested wood rot fungi, 5 were selected by resistance test on different concentration of trans,trans-fanesol. Biotransformation was performed with selected wood rot fungi on liquid culture. The metabolites detected by GC-MS analysis were nerolidol for Laetiporus sulphureus var. miniatus (jungh) Imaz and eicosane for Coriolus versicolor (L.Fr) Prlar and isoborneol for Fomitopsis pinicola and isocyclocitral for Lampteromyces japonicas. As the results, wood rot fungi could be potential biocatalyst for biotransformation of sesquiterpenes.

Biochemical Characterization of the Exopolysaccharide Purified from Laetiporus sulphureus Mycelia

The extracellular polysaccharide (EPS) was isolated from mycelial cultures of Laetiporus sulphureus var. miniatus and purified by DEAE cellulose and Sephadex G-50 column chromatography. The purified EPS (EPS-2-1) was composed of only glucose units and its molecular mass was 6.95 kDa. The chemical structure of EPS-2-1 consisted of a main chain containing (1-->4)-Glcp units with branches at the C-6 position of the chain carrying -Glcp-(1-->4)-linked residues. The effect of purified EPS on immunomodulatory genes and proteins of the Bcl-2 family was observed using cultured U937 human leukemia cells. Of note, the levels of Bax and Bad proteins treated with the EPS (4 mg/ml) were approximately 23- and 18-times higher than those in non-treated cells, respectively. These results may suggest that the EPS purified from the mushroom L. sulphureus is associated with the activation of immunomodulatory mediators, Bax and Bad proteins.

Exopolysaccharide of Laetiporus sulphureus var. miniatus downregulates LPS-induced production of NO, PGE2, and TNF-α in BV2 microglia cells via suppression of the NF-κB pathway

Our previous study showed that the exopolysaccharide (EPS) of Laetiporus sulphureus var. miniatus was well characterized and prevented cell damage in streptozotocin-induced apoptosis. However, little is known about the molecular mechanisms underlying its anti-inflammatory effects. Therefore, we attempted in this study to determine whether EPS induces a significant inhibition of pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated murine BV2 microglia cells. Our results showed that EPS significantly inhibited LPS-induced pro-inflammatory mediators, such as nitric oxide (NO), prostaglandin E2 (PGE2), and tumor necrosis factor-α (TNF-α), without any significant cytotoxicity. EPS also downregulated mRNA and protein expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF-α in LPS-induced BV2 microglia cells. Our data also revealed that EPS treatment significantly reduced translocation of nuclear factor-κB (NF-κB) subunit p65 and its DNA-binding activity in LPS-stimulated BV2 microglia cells. Furthermore, we confirmed by using proteasome inhibitor N-acetyl-l-cysteine (NAC), that the inhibition of NF-κB activity influenced the expression of pro-inflammatory genes in LPS-induced BV2 microglia cells. As expected, NAC suppressed the expression of iNOS, COX-2, and TNF-α by blocking proteasome-mediated degradation. Taken together, our data indicate that EPS inhibits the expression of pro-inflammatory mediators by suppressing NF-κB activity.

Initial acidic pH is critical for mycelial cultures and functional exopolysaccharide production of an edible mushroom, Laetiporus sulphureus var. miniatus JM 27

We conducted a time course experiment on mycelial cultures of Laetiporus sulphureus var. miniatus. The strain showed significant survival in an initial pH range of 2.0 to 7.0 for 24 days, during which time oxalic acid was accumulated. A structural analysis of purified exopolysaccharide suggested that it contained 96.1% glucose, and the mode of linkage was mainly → 4-Glcp-(1 → units, with branches at the C-6 position consisting of a Glcp-(1 → 4) linked side chain. An exopolysaccharide purified from the acidophilic strain was added to cultured U937 cells, resulting in significantly increased transcription levels of p53 and p21 genes.

Hypoglycemic effect of polysaccharides produced by submerged mycelial culture of Laetiporus sulphureus on streptozotocininduced diabetic rats

The hypoglycemic effect of the crude extracellular polysaccharides (EPS) produced from submerged mycelial culture of an edible mushroom Laetiporus sulphureus var. miniatus in streptozotocin (STZ)-induced diabetic rat was investigated. Hypoglycemic effect of EPS was evaluated in STZ-induced diabetic rats, and its possible mechanism was suggested by the results of western blot analysis and immunohistochemical staining. The results revealed that orally administrated EPS, when given 48 h after STZ treatment exhibited an excellent hypoglycemic effect, lowering the average plasma glucose level in EPS-fed rats to 43.5% of STZ-treated rats. The plasma levels of total cholesterol and triglyceride were significantly increased upon STZ treatment and they were markedly reduced by oral administration of EPS to near-normal levels. The results of immunohistochemical staining of the pancreatic tissues showed that EPS treatment considerably increased the insulin antigenesity of diabetic islet β-cells, suggesting the possibility of β-cell proliferation or regeneration by EPS therapy. Moreover, immunoblotting study revealed that protein levels of iNOS was increased and SOD2, catalase, GPx were significantly increased after EPS treatments, suggesting alleviated oxidative stress mediated by STZ. Orally administrated EPS exhibited considerable hypoglycemic effect in STZ-induced diabetic rats and that these EPS may be useful for the management of diabetes mellitus.

Relationships among three Japanese Laetiporus taxa based on phylogenetic analysis and incompatibility tests

Relationships among three Japanese Laetiporus taxa (“L. sulphureus var. sulphureus” auct. jap., L. sulphureus var. miniatus, and L. versisporus) were assessed with phylogenetic analysis and incompatibility tests. Gene phylogenies inferred from the internal transcribed spacer region of nuclear ribosomal DNA, elongation factor 1α, and β-tubulin gene regions suggested that Japanese Laetiporus was divided into four groups: the yellow pore form of L. sulphureus var. miniatus, the white pore form of L. sulphureus var. miniatus, and two “L. sulphureus var. sulphureus”/L. versisporus groups. A morphologically distinct species, Laetiporus versisporus, sharing a clade with “L. sulphureus var. sulphureus” auct. jap., was proved to be an anamorphic form of “L. sulphureus var. sulphureus” auct. jap. The “sulphureus/ versisporus” isolates showed two divergent sequence types in each region. Some isolates had intraindividual polymorphism assigned to both sequence types. This finding suggests that speciation via hybridization is ongoing in the “sulphureus/versisporus” group. Single spore isolates from the “sulphureus/versisporus” group, white pore group, and yellow pore group were incompatible with each other. Our results provided strong support for the new recognition of three Laetiporus taxa in Japan.

Production of extracellular polysaccharides by submerged mycelial culture of Laetiporus sulphureus var. miniatus and their insulinotropic properties

In the present study, optimum culture conditions for the production of extracellular polysaccharides (EPS) in submerged culture of an edible mushroom, Laetiporus sulphureus var. miniatus and their stimulatory effects on insulinoma cell (RINm5F) proliferation and insulin secretion were investigated. The maximum mycelial growth (4.1 g l(-1)) and EPS production (0.6 g l(-1)) in submerged flask culture were achieved in a medium containing 30 g l(-1) maltose, 2 g l(-1) soy peptone, and 2 mM MnSO(4).5H2O at an initial pH 2.0 and temperature 25 degrees C. In the stirred-tank fermenter under optimized medium, the concentrations of mycelial biomass and EPS reached a maximum level of 8.1 and 3.9 g l(-1), respectively. Interestingly, supplementation of deep sea water (DSW) into the culture medium significantly increased both mycelial biomass and EPS production by 4- and 6.7-fold at 70% (v/v) DSW medium, respectively. The EPS were proved to be glucose-rich polysaccharides and were able to increase proliferation and insulin secretary function of rat insulinoma RINm5F cells, in a dose-dependent manner. In addition, EPS also strikingly reduced the streptozotocin-induced apoptosis in RINm5F cells indicating the mode of the cytoprotective role of EPS on RINm5F cells.

A benzofuran glycoside and an acetylenic acid from the fungus Laetiporus sulphureus var. miniatus.

A new benzofuran glycoside, masutakeside I (1) and a new C10 acetylenic acid, masutakic acid A (2) were isolated from the fruiting bodies of the fungus Laetiporus sulphureus var. miniatus. Their structures were established by spectroscopic and chemical methods. The known compounds 3-6 were also obtained and identified as egonol, demethoxyegonol, egonol glucoside and egonol gentiobioside. Some of these compounds exhibited cytotoxicity against Kato III cells.