Diversity of Wood-Inhabiting Polyporoid and Corticioid Fungi in Odaesan National Park, Korea

Polyporoid and corticioid fungi are among the most important wood-decay fungi. Not only do they contribute to nutrient cycling by decomposing wood debris, but they are also valuable sources for natural products. Polyporoid and corticioid wood-inhabiting fungi were investigated in Odaesan National Park. Fruit bodies were collected and identified based on morphological and molecular analyses using 28S and internal transcribed spacer regions of DNA sequences. As a result, a total of 149 species, 69 genera, 22 families, and 11 orders were recognized. Half (74 species) of the species were polypores, and the other half (75 species) were corticioid fungi. Most of the species belonged to Polyporales (92 species) followed by Hymenochaetales (33 species) and Russulales (11 species). At the genus level, a high number of species was observed from Steccherinum, Hyphodontia, Phanerochaete, Postia, and Trametes. Concerning distribution, almost all the species could be found below 1,000 m, and only 20% of the species were observed from above 1,000 m. Stereum subtomentosum, Trametes versicolor, T. hirsuta, T. pubescens, Bjerkandera adusta, and Ganoderma applanatum had wide distribution areas. Deciduous wood was the preferred substrate for the collected species. Sixty-three species were new to this region, and 21 species were new to Korea, of which 17 species were described and illustrated.

Molecular characterization of a novel thermostable laccase PPLCC2 from the brown rot fungus Postia placenta MAD-698-R

Background Laccase has been considered important for the degradation of lignocellulose by wood rot fungi. The properties and functions of laccase in white rot fungi have been investigated extensively, but those from brown rot fungi remain largely unknown. In this paper, a laccase isoform Pplcc2 from the brown rot fungus Postia placenta MAD-698-R was expressed heterologously in Pichia pastoris GS115, purified and the properties of the enzyme were determined. Results The molecular weight of the protein was determined to be 67 kDa using SDS-PAGE. It cannot oxidize syringaldazine (SGZ), but it can oxidize 2,2'-azino-di-(3-ethylbenzothialozin-6-Sulfonic acid) (ABTS) and 2,6-dimethoxyphenol (DMP). Specific activity for ABTS was 1960 ± 19 Unit/mg. The catalytic constant (k cat) was 1213 ± 18.3 s-1 for ABTS and 293.2 ± 21.9 s-1 for DMP. Km was 22.08 µM for ABTS and 11.62 µM for DMP. The optimal pH for the oxidation of ABTS and DMP was 3.5 and 5.0 respectively. The optimal temperature for the oxidation of ABTS and DMP was 60°C. Conclusions This is the first identified thermo activated and thermostable laccase in brown rot fungi. This investigation will contribute to understanding the roles played by laccases in brown rot fungi.

Enhancement of beta-Glucosidase Activity from a Brown Rot Fungus Fomitopsis pinicola KCTC 6208 by Medium Optimization

beta-Glucosidase, which hydrolyzes cellobiose into two glucoses, plays an important role in the process of saccharification of the lignocellulosic biomass. In this study, we optimized the activity of beta-glucosidase of brown-rot fungus Fomitopsis pinicola KCTC 6208 using the response surface methodology (RSM) with various concentrations of glucose, yeast extract and ascorbic acid, which are the most significant nutrients for activity of beta-glucosidase. The highest activity of beta-glucosidase was achieved 3.02% of glucose, 4.35% of yeast extract, and 7.41% ascorbic acid where ascorbic acid was most effective. The maximum activity of beta-glucosidase predicted by the RSM was 15.34 U/mg, which was similar to the experimental value 14.90 U/mg at the 16th day of incubation. This optimized activity of beta-glucosidase was 23.6 times higher than the preliminary activity value, 0.63 U/mg, and was also much higher than previous values reported in other fungi strains. Therefore, a simplified medium supplemented with a cheap vitamin source, such as ascorbic acid, could be a cost effective mean of increasing beta-glucosidase activity.

Taxonomic Study of the Genus Abundisporus in Korea

The polypore genus Abundisporus Ryvarden is characterized by resupinate to pileate fruitbodies with a purplish brown hymenophore, slightly thick-walled, pale yellowish and non-dextrinoid basidiospores, and causing white rot. A purple color hymenophore, an easily observable and striking character, was considered the main distinctive feature at the generic level within polypores. However, due to highly similar basidiocarp features, species identification within these purple polypores is particularly difficult. Three species of purple colored polypores have been reported in Korea (Abundisporus fuscopurpureus, A. pubertatis, and Fomitopsis rosea). Based on morphological re-examination, ecological information, and sequence analysis of the internal transcribed spacer, we showed that previous classification was incorrect and there is only one species (A. pubertatis) in Korea. We provide a detailed description of A. pubertatis in Korea, as well as a taxonomic key to distinguish wood rot fungi with a purple hymenophore.

Methyl 9-Oxo-(10E,12E)-octadecadienoate Isolated from Fomes fomentarius Attenuates Lipopolysaccharide-Induced Inflammatory Response by Blocking Phosphorylation of STAT3 in Murine Macrophages

Fomes fomentarius is a fungus of the Polyporaceae family and is used in traditional oriental therapies. Although the anti-inflammatory activities of this species have been previously reported, the identity of the bioactive compounds responsible for this activity remains unknown. Here, we investigated whether methyl 9-oxo-(10E,12E)-octadecadienoate (FF-8) purified from F. fomentarius exerts anti-inflammatory activity in murine macrophages stimulated with lipopolysaccharide (LPS). FF-8 suppressed secretion of nitric oxide (NO) and prostaglandin E2 through downregulation of inducible NO synthase and cyclooxygenase-2 expression induced by LPS. In addition, pretreatment of cells with FF-8 led to a reduction in levels of secreted inflammatory cytokines such as tumor necrosis factor-alpha and interleukin-6 in macrophages stimulated with LPS. Conversely, FF-8 did not affect nuclear factor kappaB, p38, c-Jun NH2-terminal kinase, and extracellular signal-regulated kinase pathways. Instead, FF-8 specifically interfered with signal transducer and activator of transcription 3 (STAT3) phosphorylation induced by LPS. Collectively, this study demonstrated that FF-8 purified from F. fomentarius suppresses inflammatory responses in macrophages stimulated with LPS by inhibiting STAT3 activation. Further studies will be required to elucidate the anti-inflammatory effect of FF-8 in vivo.

Differential Modulation of Lipopolysaccharide-Induced Inflammatory Cytokine Production by and Antioxidant Activity of Fomentariol in RAW264.7 Cells

Medicinal mushrooms have been used worldwide to treat cancer and modulate the immune system. Over the last several years, there has been increasing interest in isolating bioactive compounds from medicinal mushrooms and evaluating their health beneficial effects. Fomes fomentarius is used in traditional oriental medicine and is known to possess antioxidant, anti-inflammatory, antidiabetic, and antitumor effects. In the present study, we isolated fomentariol from Fomes fomentarius and investigated its anti-inflammatory effect in murine macrophages (RAW264.7 cells) stimulated with lipopolysaccharides. Fomentariol inhibited the production of nitric oxide and intracellular reactive oxygen species triggered by lipopolysaccharides. Interestingly, fomentariol differentially regulated cytokine production triggered by lipopolysaccharides. Fomentariol effectively suppressed the production of interleukin-1beta and interleukin-6 but not tumor necrosis factor-alpha. The inhibitory effect of fomentariol against nitric oxide, interleukin-1beta, and interleukin-6 production was possibly mediated by downregulation of the extracellular signal-regulated kinase signaling pathway. Taken together, our results suggest that fomentariol differentially modulated inflammatory responses triggered by lipopolysaccharides in macrophages and is one of the bioactive compounds that mediate the physiological effects of Fomes fomentarius.

Grape stalks as substrate for white rot fungi, lignocellulolytic enzyme production and dye decolorization / Uso del escobajo como sustrato para el crecimiento de hongos de la pudrición blanca, la producción de enzimas ligninolíticas y la decoloración de tinturas

The aim of this work was to evaluate the potential of grape stalks, an agroindustrial waste, for growth and lignocellulolytic enzyme production via solid-state fermentation, using the following three white rot fungi: Trametes trogii, Stereum hirsutum and Coriolus antarcticus. The decolorization of several dyes by the above mentioned cultures was also investigated. Similar values of dry weight loss of the substrate were measured after 60 days (33-43 %). C. antarcticus produced the highest laccase and Mn-peroxldase activities (33.0 and 1.6 U/g dry solid). The maximum endoglucanase production was measured in S. hirsutum cultures (10.4 U/g), while the endoxylanase peak corresponded to T. trogii (14.6 U/g). The C. antarcticus/grape stalk system seems potentially competitive in bioremediation of textile processing effluents, attaining percentages of decolorization of 93, 86, 82, 82, 77, and 58 % for indigo carmine, malachite green, azure B, remazol brilliant blue R, crystal violet and xylidine, respectively, in 5 h.

El objetivo de este trabajo fue evaluar el potencial del escobajo, un residuo agroindustrial, como sustrato para el crecimiento y la producción de enzimas lignocelulósicas de tres hongos causantes de pudrición blanca en la madera: Trametes trogii, Stereum hirsutum y Coriolus antarcticus. Para ello se utilizaron técnicas de fermentación en estado sólido. También se ensayó la decoloración de colorantes industriales sobre estos cultivos. La pérdida de peso seco del sustrato fue similar después del día 60 (33-43 %). C. antarcticus produjo las mayores actividades de lacasa y Mn-peroxidasa (33,0 y 1,6 U/g peso seco). La mayor actividad endoglucanasa fue medida en cultivos de S. hirsutum (10,4 U/g), y la mayor actividad endoxilanasa en T. trogii (14,6 U/g). El sistema C. antarcticus/escobap mostró un importante potencial para su aplicación en la biorremediación de efluentes textiles, con porcentajes de decoloración de 93, 86, 82, 82, 77 y 58 % para índigo carmín, verde de malaquita, azure B, azul R brillante de remazol, cristal violeta y xilidina, respectivamente, en 5 h.

Fomitoside K, a New Lanostane Triterpene Glycoside from the Fruiting Body of Fomitopsis nigra

In an effort to identify the chemical constituents of fruiting bodies of Fomitopsis pinicola, a new lanostane triterpene glycoside, designated as fomitoside K, has been isolated from its methanolic extract. Its chemical structure was assigned on the basis of various spectroscopic studies.

Isolation and characterization of a novel lignocellulose decomposing fungal strain.

A strain F1 with high cellulase activity obtained from the deadwood stack was characterized as Ceriporia lacerate by examination of the general taxonomical characteristics and phylogenetic sequence analysis of rDNA ITS gene. The endoglucanase (EG) and filter paper cellulase (FPase) activities of the strain showed remarkable stability in the pH range of 4.0-7.0, and maintained about their maximal value of 76% and 50% after incubation at 70˚C for 6 h respectively. The strain grew particularly well with CMC-Na (1.0%) and yeast extract (0.4%) at 28˚C (pH 6.0) in flasks stirred at 150 × g for 6 days. Based on the thermostability and pH stability of cellulase, the strain appears to have potential in industrial applications and bioresource utilization.

Extraction and characterization of polygalacturonase of Fomes sclerodermeus produced by solid-state fermentation / Extracción y caracterización de poligaracturonasa de Fomes sclerodermeus producida por fermentación en estado sólido

Polygalacturonase (PG) production by Fomes sclerodermeus using solid-state fermentation (SSF) was carried out. Maximal PG activity (26 U/gdw) was obtained between days 11 and 13 at the end of exponential growth. PG activity in the crude extract was more stable at pH 5-6 and 30 °C and had optimum activity at pH 5 and 50 °C. Optimal conditions for PG extraction were: one time extraction with Na2SO4 as solvent with 10 min. of agitation. In a scale-up system, PG activity per gram of dry substrate decreased about 60% compared with the activity obtained in an Erlenmeyer flask; however, high total PG activity was obtained.

Se estudió la producción de poligalacturonasa (PG) por Fomes sclerodermeus usando técnicas de fermentación en estado sólido. La actividad PG máxima (26 U/g ps) fue observada entre los días 11 y 13. La actividad PG en los extractos crudos fue más estable a pH 5-6 y 30 °C, con una actividad óptima a pH 5 y a 50 °C. Las condiciones óptimas para la extracción de PG se lograron con una única extracción empleando Na2SO4 como solvente, con 10 minutos de agitación. En el escalado del sistema, la actividad PG por gramo de peso seco de sustrato disminuyó cerca de 60% comparada con la obtenida en frascos Erlenmeyer, pero la actividad total fue mayor.

Checklist of Hymenomycetes (Aphyllophorales s.l.) and Heterobasidiomycetes in Israel

A checklist is presented concerning the species composition of Hymenomycetes (Aphyllophorales s.l.) and Heterobasidiomycetes in Israel based on data of previous studies and field sample collections. In total, 242 species are presented, of which five are new records for the Israeli mycobiota, namely Australohydnum dregeanum, Ceriporiopsis consobrina, C. resinascens, Fibroporia vaillantii, and Postia inocybe. The distribution and habitat of each species in Israel are also summarized. This checklist will serve as valuable data for future species diversity studies of these fungi in Israel.

Immunomodulatory effects of Fomes fomentarius polysaccharides: an experimental study in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the immunomodulatory effects of Fomes fomentarius polysaccharides (FFP) in mice.</p><p><b>METHODS</b>MTT assay was employed to evaluate the in vitro metabolic activity of the mouse splenocytes treated with FFP at different concentrations, and the secretion of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (INF-gamma) and interleukin 2 (IL-2) from the cells were measured by enzyme-linked immunosorbent assay. The changes in the phagocytotic activity of mouse macrophage in response to FFP treatment were evaluated by phagocytosis percentage of chicken red blood cells (CRBCs). The effect of FFP on the humoral immunity was assessed in mice immunized with sheep red blood cells (SRBCs) by measuring the serum levels of specific antibody (hemolysin) against SRBCs.</p><p><b>RESULTS</b>FFP at the concentrations of 25, 50, and 100 microg/ml all significantly enhanced the metabolic activity of mouse splenocytes in vitro and increased the production of TNF-alpha, IFN-gamma and IL-2. FFP treatment also markedly enhanced the metabolic activity of mouse peritoneal exudate cells and TNF-alpha production by the cells. At the doses of 25, 50, and 100 mg/kg, FFP significantly increased serum hemolysin level in mice immunized with SRBCs, and FFP at 50 and 100 mg/kg obviously increased the capacity of mouse peritoneal macrophages in vivo for CRBC phagocytosis.</p><p><b>CONCLUSION</b>FFP can promote the secretion of TNF-alpha, IFN-gamma and IL-2 by mouse immunocytes and enhance mouse humoral immune response and the phagocytotic activity of the macrophages.</p>

First Report of Ceriporiopsis resinascens (Phanerochaetaceae, Basidiomycota) in Korea

An unrecorded Ceriporiopsis species was collected at Mt. Gariwang, Gangwon Province, in 2008. Based on morphological characteristics, such as a fully resupinate basidiocarp, a reddish white to pinkish poroid hymenophore and a monomitic hyphal system with clamp connections, the species was identified as Ceriporiopsis resinascens. This is the first report of Ceriporiopsis resinascens in Korea. We confirmed the identity of the species as Ceriporiopsis resinascens based on ITS sequence analysis.

Degradation of Three Aromatic Dyes by White Rot Fungi and the Production of Ligninolytic Enzymes

This study was conducted to evaluate the degradation of aromatic dyes and the production of ligninolytic enzymes by 10 white rot fungi. The results of this study revealed that Pycnoporus cinnabarinus, Pleurotus pulmonarius, Ganoderma lucidum, Trametes suaveolens, Stereum ostrea and Fomes fomentarius have the ability to efficiently degrade congo red on solid media. However, malachite green inhibited the mycelial growth of these organisms. Therefore, they did not effectively decolorize malachite green on solid media. However, P. cinnabarinus and P. pulmonarius were able to effectively decolorize malachite green on solid media. T. suaveolens and F. rosea decolorized methylene blue more effectively than any of the other fungi evaluated in this study. In liquid culture, G. lucidum, P. cinnabarinus, Naematoloma fasciculare and Pycnoporus coccineus were found to have a greater ability to decolorize congo red. In addition, P. cinnabarinus, G. lucidum and T. suaveolens decolorized methylene blue in liquid media more effectively than any of the other organisms evaluated in this study. Only F. fomentarius was able to decolorize malachite green in liquid media, and its ability to do so was limited. To investigate the production of ligninolytic enzymes in media containing aromatic compounds, fungi were cultured in naphthalene supplemented liquid media. P. coccineus, Coriolus versicolor and P. cinnabarinus were found to produce a large amount of laccase when grown in medium that contained napthalene.

Hypoglycemic Effects of Exo-biopolymers Produced by Five Different Medicinal Mushrooms in STZ-induced Diabetic Rats

Hypoglycemic effects of exo-biopolymers (EBP) produced by submerged mycelial cultures of Coriolus versicolor, Cordyceps sinensis, Paecilomyces japonica, Armillariella mellea, and Fomes fomentarius were investigated in streptozotocin (STZ)-induced diabetic rats. The rats from each experimental group were orally administered with EBPs (100 mg/kg BW) daily for 2 weeks. Though the hypoglycemic effect was achieved in all the cases, however, C. versicolor EBP proved as the most potent one. The administration of the C. versicolor EBP substantially reduced (29.9%) the plasma glucose level as compared to the saline administered group (control). It also reduced the plasma total cholesterol (TC), triglyceride (TG), aspartate aminotransferase (AST) and, alanine aminotransferase (ALT) levels by 9.22, 23.83, 16.93, and 27.31%, respectively. The sugar and amino acid compositions of this EBP were also analyzed in detail.

Anti-complementary Activities of Exo- and Endo-biopolymer Produced by Submerged Mycelial Culture of Eight Different Mushrooms

The Elfvingia applanata (EA), Hericium erinaceum (HE),Grifola frondosa (GF), Pholiota nameko (PN), Pleurotus eryngii (PE), Trametes suaveolens (TS), Fomes fomentarius (FF), and Inonotus obliquus (IO) could produce the endo- (EN) and exo-biopolymer (EX) in submerged culture. The highest anti-complementary activity of the EN was exhibited by PN (49.1%), followed by HE (38.6%), TS (37.0%),and FF (33.0%),whereas the high activity of the EX was found with GF (59.8%),followed by HE (36.3%),TS (30.8%),and IO (28.8%). The EN of P. nameko (EN-PN) and EX of G. frondosa (EX-GF) were found to contain 78.6% and 41.2% carbohydrates, while 21.4% and 58.8% protein, respectively. The sugar and amino acid compositions of EN-PN and EX-GF were also analyzed in detail.

Evaluation of Prognostic Factors in Acute Renal Failure at the Emergency Department

PURPOSE: This study intended to identify prognostic factors influencing recovery, progressing chronic renal failure (CRF) and mortality in acute renal failure (ARF) patients at the emergency department. METHODS: We retrospectively analyzed 104 patients with ARF and with serum creatinine (Cr) level above 2 mg/dL and glomerular filtration rate (GFR) 60 years, underlying disease, cause of ARF, urine volume, existence of oliguria, duration of oliguria, proteinuria, GFR, serum BUN, serum Cr, BUN/Cr ratio, arterial pH, PaO2, S a O2, serum Na+, serum K+, serum albumin, serum total bilirubin, serum osmolarity, serum AST, serum ALT, serum creatine kinase, and serum myoglobin were all significant factors discriminating between recovery patients and nonrecovery patients (progressing CRF, mortality). Based on a multivariate analysis, sex, existence of oliguria, duration of oliguria, GFR, BUN/Cr ratio, PaO2, serum K+, SaO2, serum bilirubin, and serum osmolarity were useful factors which might affected non-recovery. CONCLUSION: In ARF, the prognostic factors were serum K+, S a O2, duration of oliguria, and BUN/Cr ratio. The higher serum K+, the lower SaO2, the longer the duration of oliguria, and the lower the BUN/Cr ratio are for ARF patients at the emergency department, the more the intensive care emergency physician must perform.

Characterization About Vibrio alginolyticus Phage Isolated from Marine Products

Two phages for the pathogenic V. alginolyticus were isolated from marine products. These 2 phages were examined temperature stability, pH stability, inactivation by UV irradiation, damage on restriction system of host cell, antibody production, structure protein analysis and western blotting assay. V. alginolyticus phages(VAPs) fomed plaques about 0.5 - 0.9mm in diameter and bands 50 - 60% in sucrose density gradient, VAPs were stable below 65'C, pH 5 - 10 and mostly inactivation by UV irradiation for 120sec. Latend period was 15 - 20 min. and burst size was 1.3 - 1.4 * 10 PFU/cell. Restriction system of V. alginolyticus isolated was mostly inactivated by 45C, 20min. heating. VAPs had 14 specific structural proteins and 5 proteins related to antibody production.

Electromicroscopic Characterization about Vibrio alginolyticus Phage Isolated from Marine Products

The study of bacteriophage began by F.W. Twort in 1915 and the lytic cycle recognized by d'Herellel in 1917. It repeated about the marine bacteriophage containing Vibrio phage by Smith, Spencer and Ju. Authors isolated 2 virulent phages for the pathogenic V. alginolyticus from marine products. These 2 phages were examined their ultrastructure & host-infection by elecron microscopy and in vivo test using skin of rats. V. alginolyticus phages(VAPs) fomed plaques about 0.5 - 0.9mm in diameter and bands 50 - 60% in sucrose density gradient. VAP had 50 - 120nm tail and 40 - 90nm head in diameter. In vivo test, using rat skin, as well as in vitro test VAP had the activity to V. alginolyticus isolated.