Phosphatidylcholine and ceramide derivatives from white rot fungus Microporus xanthropus PP17-20.

The undescribed phosphatidylcholine (1), along with twelve known compounds, was isolated from the cultures of white rot fungus Microporus xanthropus PP17-20. In this work the fungus was cultivated in Yeast-Malt extract medium to explore active compound production. The chemical structures were elucidated on the basis of spectroscopic and HRESIMS data. Several isolated compounds were evaluated for anti-proliferative activity against A549 and MCF-7 cancer cell lines.

Engleromophilane, a new broad spectrum bioactive eremophilane-type sesquiterpene from Engleromyces sinensis fungus.

A new eremophilane sesquiterpene, named engleromophilane (1) together with known eremoxylarin E (2) and steroids (3-7) were isolated from the fungus Engleromyces sinensis culture. The structures were deduced by the analysis of spectroscopic and MS data, together with the comparison of calculated 13C NMR chemical shifts and Electronic Circular Dichroism (ECD) spectra. Compound 1 showed cytotoxic effects against Hela, PC-3, HT29 and A549 cell lines with IC50 in the ranges of 4.84-9.48 µg/mL. Compounds 1 and 2 exhibited substantial antimicrobial activity against E. coli, S. aureus, and B. subtilis. Moreover, compounds 1-3 showed α-glucosidase inhibitory activity, in which 2 displayed a strong inhibitory effect with an IC50 value of 0.13 ± 0.01 µg/mL. This work has given additional value to the E. sinensis fungus as a remarkable bioactive compound producer, together with the possibility of increasing cultivation to industrial scales.

Antiproliferative polyketides from fungus Xylaria cf. Longipes SWUF08-81 in different culture media.

Bioactive compounds from the wood-decay fungus Xylaria cf. longipes SWUF08-81, cultivated in three different culture media (GM, YM and PDB), were isolated. Their structures and stereochemistry were deduced from spectroscopic and MS data analysis, together with quantum chemical calculations of 13C NMR chemical shifts and electronic circular dichroism (ECD) spectra. Five undescribed polyketides including dibenzofuran (1), mellein (2), dihydroisocoumarin (15), and two pyrans (16, 17), together with twenty-three compounds were determined. Compounds 18 and 20 were significantly toxic against cancer cell lines (HCT116, HT29, MCF-7 and HeLa) based on the MTT assay. Quantification by HPLC showed that 18 was produced three-fold higher in the broth of PDB than YM. These studies showed that the production of different compounds were primarily dependent on nutrition sources and it has given a starting point for the growth optimization conditions for the scaling up of bioactive compounds production.

Screening of White-Rot Fungi Isolates for Decolorization of Pulp and Paper Mill Effluent and Assessment of Biodegradation and Biosorption Processes.

Ten white-rot fungal isolates were evaluated for the decolorization potential of pulp and paper mill effluent. Trametes elegans PP17-06, Pseudolagarobasidium sp. PP17-33, and Microporus sp.2 PP17-20 showed the highest decolorization efficiencies between 42 and 54% in 5 d. To reveal the mechanisms involved in decolorization and assess the long-term performance, PP17-06, which showed the highest decolorization efficiency, was further investigated. It could reduce the ADMI color scale by 63.6% in 10 d. However, extending the treatment period for more than 10 d did not significantly enhance the decolorization efficiencies. The maximum MnP activity of 3.27 U L-1 was observed on the 6 d during the biodegradation. In comparison, laccase activities were low with the maximum activity of 0.38 U L-1 (24 d). No significant LiP activities were monitored during the experiment. Dead fungal biomass showed an optimum decolorization efficiency of 44.18% in 8 d employing the biosorption mechanism. No significant changes in the decolorization efficiency were observed after that, suggesting the equilibrium status was reached. These results revealed that PP17-06 has the potential to decolorize pulp and paper mill effluent by employing both biodegradation and biosorption processes.

Isolation, Characterization, and Comparative Genomics of the Novel Potential Probiotics from Canine Feces.

Lactic acid bacteria (LAB) are commonly used as probiotics; however, not all LAB strains have the same beneficial effects. To successfully use LAB as probiotics in canines, LAB species should originate from the canine intestinal tract as they display host specificity. The objective of this study was to investigate the phenotypic and genomic traits of potential probiotic LAB isolated from canine fecal samples. Twenty LAB samples were evaluated for their potential probiotic characteristics including resistance to low pH, bile salts, hydrophobicity, auto-aggregation, co-aggregation, adhesion to epithelia or mucosa, and production of inhibitory compounds. Additionally, we evaluated their safety and other beneficial effects on canine health, such as DPPH free radical scavenging, and ß-galactosidase. Four strains demonstrated potential probiotic characteristics and were selected: Enterococcus hirae Pom4, Limosilactobacillus fermentum Pom5, Pediococcus pentosaceus Chi8, and Ligilactobacillus animalis FB2. Safety evaluations showed that all strains lacked hemolytic activity, could not produce biogenic amines, and did not carry any pathogenic genes. In addition, L. fermentum Pom5 and P. pentosaceus Chi8 displayed susceptibility to all antibiotics and concordant with the absence of antibiotic resistance genes. Based on their phenotypic and genomic characteristics, L. fermentum Pom5 and P. pentosaceus Chi8 were identified as potential probiotic candidates for canines.

Xylaria furcata reconsidered and nine resembling species.

BACKGROUND: Xylaria collections from termite nests with dichotomously branched stromata have been identified as X. furcata. However, Léveillé's original material is no longer available, and the modern interpretation of X. furcata is based on a 1908 collection made by von Höhnel from termite nests at Buitenzorg Botanical Garden in Java. A packet of this von Höhnel material at FH was designated as the neotype by Rogers et al. in 2005. RESULTS: We reexamined the neotype from FH and its duplicates from various herbaria and found that three different species were mixed in these specimens. Despite that all of them have dichotomously branched stromata and tiny ascospores, only one fits the 2005 neotypification of X. furcata, where exposed perithecial mounds on the stromatal surface were unambiguously indicated. This portion of material is redesignated as the neotype, while the other two species with immersed perithecia are described as new: X. hoehnelii and X. robustifurcata. The ITS sequence obtained from the neotype helped us designate a specimen with cultures obtained from it as the epitype. From specimens identifiable as X. furcata, we describe four new species: X. brevifurcata, X. furcatula, X. insignifurcata, and X. tenellifurcata. Additionally, we recognize X. furcata var. hirsuta at the species level as X. hirsuta and consider X. scoparia a distinct species rather than a synonym of X. furcata. Molecular phylogenetic analyses based on three protein-coding loci showed that X. furcata and resembling species were grouped into two clusters: the X. furcata cluster with half-exposed to fully exposed perithecial mounds and the X. hoehnelii cluster with largely immersed perithecial mounds. CONCLUSION: Ten species are recognized for X. furcata and resembling species, all of which could have been identified as X. furcata in the past. Its diversity has been overlooked primarily due to the small and similar stromata. Several additional species have been confirmed to be related to X. furcata by DNA sequences but are yet to be described due to the lack of mature stromata. While the species diversity of macrotermitine termites is equally high in Africa as in Asia, all of the species are primarily found in Asia, with X. hirsuta as the only exception. This suggests that there may be many more undiscovered species for this fungal group.

Exploring the Xylariaceae and its relatives.

The Xylariaceae and its relatives rank as one of the best-known members of the Ascomycota. They are now well recognized for their diversity, global distribution, ecological activities and their outstanding novel metabolites with wide ranging bioactivity.

Hypoxylonone, a new oxa-bridged seven-membered ring analog from fungus Hypoxylon cf. subgilvum SWUF15-004.

A new oxa-bridged seven-membered ring analog, hypoxylonone (1), and thirteen known compounds (2-14) were isolated from fungus Hypoxylon cf. subgilvum SWUF15-004. The structures were elucidated by the analysis of spectroscopic (IR, 1 D and 2 D NMR), HRESIMS and X-ray diffraction (MoKα) data. Several isolated compounds were evaluated for cytotoxicity against four human cancer cell lines (HeLa, HT29, MCF-7, A549). Compound 1 exhibited weak inhibitory effects of the nitric oxide production in RAW264.7 cells. Compounds 8 and 9 exhibited slight cytotoxicity.

High Potential Decolourisation of Textile Dyes from Wastewater by Manganese Peroxidase Production of Newly Immobilised Trametes hirsuta PW17-41 and FTIR Analysis.

Coloured wastewater from the textile industry is a very serious global problem. Among 16 different white-rot fungal isolates, Trametes hirsuta PW17-41 revealed high potential for decolourisation of mixed textile dyes (Navy EC-R, Ruby S3B and Super Black G) from real industrial wastewater samples. The efficiency of dye decolourisation was evaluated using the American Dye Manufacturers' Institute (ADMI) standard methodology. The suitable support for fungal mycelium immobilisation was nylon sponges. The optimal dye decolourisation (95.39%) was achieved by using palm sugar and ammonium nitrate as carbon and nitrogen sources, respectively. The initial pH was 5 and the agitation speed was 100 rpm at 30 °C. The ADMI values of textile dyes decreased from 2475 to 114 within two days, reducing the treatment time from seven days before optimisation. The major mechanism of dye decolourisation was biodegradation, which was confirmed by UV-visible and FTIR spectra. Manganese peroxidase (MnP) (4942 U L-1) was found to be the main enzyme during the decolourisation process at an initial dye concentration of 21,200 ADMI. The results indicated the strong potential of immobilised fungal cells to remove high concentrations of textile dyes from industrial wastewater and their potential ability to produce high MnP and laccase activities that can be used in further application.

New furan derivatives from Annulohypoxylon spougei fungus.

Two new furan derivatives, annulofurans A-B (1-2), together with six known compounds were isolated from Annulohypoxylon spougei fungus. The structures were determined based on NMR and mass spectrometry data. The absolute configurations of annulofurans A-B were determined by Electronic Circular Dichroism (ECD) experiment and comparisons with the experimental ECD spectra of synthesized stereoisomers. The evaluation of the effects on radish and ruzi grass radicle elongation by the isolated compounds showed that annulofuran A affected radicle elongation of ruzi grass. The known 2-hydroxyphenylacetic acid methyl ester (7) had significant effects against both radish and ruzi grass radicle elongation, which were comparable to the commercial herbicide, glyphosate.

Antioxidant Activity and Cytotoxicity against Cancer Cell Lines of the Extracts from Novel Xylaria Species Associated with Termite Nests and LC-MS Analysis.

Xylaria species associated with termite nests or soil have been considered rare species in nature and the few which have been reported upon have been found to act as a rich source of bioactive metabolites. This study evaluated 10 ethyl acetate extracts of five new Xylaria species associated with termite nests or soil for their antioxidant activity, and cytotoxicity against different cancer and normal cell lines. DPPH and ABTS radical scavenging activities of the extracts demonstrated strong capacity with low IC50 values. The highest observed activities belonged to X. vinacea SWUF18-2.3 having IC50 values of 0.194 ± 0.031 mg/mL for DPPH assay and 0.020 ± 0.004 mg/mL for ABTS assay. Total phenolic content ranged from 0.826 ± 0.123 to 3.629 ± 0.381 g GAE/g crude extract which correlated with antioxidant activities. The high total phenolic content could contribute to the high antioxidant activities. Cytotoxicity was recorded against A549, HepG2, HeLa and PNT2 and resulted in broad spectrum to specific activity depending on the cell lines. The highest activities were observed with X. subintraflava SWUF16-11.1 which resulted in 11.15 ± 0.32 to 13.17 ± 2.37% cell viability at a concentration of 100 µg/mL. Moreover, LC-MS fingerprints indicated over 61 peaks from all isolates. There were 18 identified and 43 unidentified compounds compared to mass databases. The identified compounds were from various groups of diterpenoids, diterpenes, cytochalasin, flavones, flavonoids, polyphenols, steroids and derivatives, triterpenoids and tropones. These results indicate that Xylaria spp. has abundant secondary metabolites that could be further explored for their therapeutic properties.

Anti-inflammatory and anti-proliferative activities of chemical constituents from fungus Biscogniauxia whalleyi SWUF13-085.

The fungus Biscogniauxia whalleyi SWUF13-085 from the Graphostomataceae family was studied for potential anti-inflammatory and anticancer agents. A diverse array of natural products was identified. Six of which were undescribed compounds, including xylariterpenoids L-N, (1R,2S,6R,7S)-1,2-dihydroxy-α-bisabolol, 6-[(1R)-1-hydroxy-1-methyl-2-propenyl]-4-methoxy-3-methyl-2H-pyran-2-one and (1R*,4S*,5S*,7S*,10R*)-guaia-11 (12)-en-7,10-diol. Several of the isolated compounds such as bergamotene, guaiane and phthalide derivatives showed activity in both the inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells with IC50 values in the range of 2.48-10.82 µg/mL and anti-proliferation against HeLa cells with IC50 values in the range of 8.64-31.16 µg/mL. While compounds such as cerebrosides A and C only exhibited inhibitory effects on NO production with IC50 values in the range of 4.45-10.28 µg/mL.

Twelve New Taxa of Xylaria Associated with Termite Nests and Soil from Northeast Thailand.

The diversity of Xylaria species associated with termite nests in northeast Thailand was investigated. Among the 14 taxa included in this study, 11 species and one variety were described as new, and another two species resemble the existing taxa, X. escharoidea and X. nigripes. The newly described taxa are X. chaiyaphumensis, X. conica, X. fulvescens, X. ischnostroma, X. margaretae, X. minima, X. reinkingii var. microspora, X. siamensis, X. sihanonthii, X. subintraflava, X. thienhirunae, and X. vinacea. Their morphological and cultural characteristics are described and illustrated, and their ITS, α-actin and ß-tubulin sequences were analysed. A dichotomous key to the 17 species of Xylaria occurring in Thailand is provided.

A new α-pyrone derivative from Annulohypoxylon stygium SWUF09-030.

A new α-pyrone derivative, namely annulopyronoate (1), together with the known isobenzofuranone, diphenyl ether, sesquiterpene, phenolic and diketopiperazine derivatives (2-12) were isolated from a filamentous fungus Annulohypoxylon stygium SWUF09-030. The structures were determined by analysis of spectroscopic and MS data. The absolute configuration of 1 was determined by comparison of experimental and calculated Electronic Circular Dichroism (ECD) spectra. Compounds 3-5 and 7 exhibited both anti-proliferative effects against HeLa, HT29, HCT116, MCF-7 and Vero cell lines and inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells.

Anti-inflammatory and cytotoxic agents from Xylaria sp. SWUF09-62 fungus.

The ongoing search for anti-cancer agents from microorganisms led to the isolation of four new compounds including 6-ethyl-8-hydroxy-4H-chromen-4-one (1), 6-ethyl-7,8-dihydroxy-4H-chromen-4-one (2), (3S)-3,4-dihydro-8-hydroxy-7-methoxy-3-methylisocoumarin (3) and (3S)-3,4-dihydro-5,7,8-trihydroxy-3-methylisocoumarin (4), together with eleven known compounds (5-15) from Xylaria sp. SWUF09-62 fungus. The chemical structures were deduced from IR, 1D and 2D NMR, and MS data. The absolute configurations of 3 and 4 were determined by ECD experiment. Compounds 2 and 4 indicated possible chemo-prevention and chemo-therapeutic properties, exhibited anti-inflammatory properties by reducing nitric oxide production in LPS-stimulated RAW264.7 cells (IC50 = 1.57 ± 0.25 and 3.02 ± 0.27 µg/mL) and cytotoxicity against HT29 cells (IC50 = 16.46 ± 0.48 and 97.78 ± 7.14 µg/mL).

Chemical constituents and cytotoxic activity from the wood-decaying fungus Xylaria sp. SWUF08-37.

A new cyclic pentapeptide, pentaminolarin (1), and a new cytochalasin, xylochalasin (2), along with thirteen known compounds (3-15) were isolated from the wood-decaying fungus Xylaria sp. SWUF08-37. The absolute configurations of 1 were determined by a combination of Marfey's method and TDDFT ECD calculation and the absolute configurations of 2 were established by TDDFT ECD calculation. Compound 12 showed moderate cytotoxicity against HeLa (IC50 = 19.60 µg/mL), HT29 (IC50 = 17.31 µg/mL), HCT116 (IC50 = 14.28 µg/mL), MCF-7 (IC50 = 15.38 µg/mL), and Vero (IC50 = 24.97 µg/mL) cell lines by MTT assay. Compounds 1 and 2 showed slight cytotoxicity against all tested cancer cell lines.

Utilization of oil palm decanter cake for valuable laccase and manganese peroxidase enzyme production from a novel white-rot fungus, Pseudolagarobasidium sp. PP17-33.

Oil palm decanter cake (OPDC) in the current study was converted to valuable products as laccase and manganese peroxidase (MnP) by an undescribed strain of the white-rot fungus, Pseudolagarobasidium sp. PP17-33. The optimization to enhance the production of enzymes through solid-state fermentation was performed using Plackett-Burman design and response surface methodology. The highest observed laccase was 5.841 U/gds and observed MnP was 5.156 U/gds, which enhanced yield by 2.59-fold and 1.94-fold from the non-optimization. The optimized medium (mg/g of OPDC) consisted of 0.852 mg CuSO4·5H2O, 13.512 mg glucose, 2 mg yeast extract, 0.2 mg KH2PO4, 1.5 mg MgSO4·7H2O, 0.01 mg FeSO4·7H2O, 0.15 mg MnSO4·H2O, 0.01 mg ZnSO4·7H2O and 0.3 mg Tween 80 (pH 5.0) when incubated at 30 °C for 7 days. The most significant variables of laccase and MnP productions were CuSO4·5H2O and glucose concentrations. This study is the first to report on the production of ligninolytic enzymes from OPDC waste using white-rot fungi. In addition, five different white-rot fungi, Coriolopsis aspera, C. retropicta, Dentipellis parmastoi, Nigroporus vinosus and Tyromyces xuchilensis, are newly observed producers of ligninolytic enzymes in Thailand. The results obtained from this study are significant not only for agro-industrial waste management but also for value-added enzyme production.

A new amino amidine derivative from the wood-decaying fungus Xylaria cf. cubensis SWUF08-86.

The secondary metabolites of Xylaria cf. cubensis SWUF08-86 fungus were investigated, and the chromatographic separation of the crude extracts yielded seventeen compounds. The structure elucidation by spectroscopic analysis including 1D and 2D NMR and the comparison of these data with literature, along with HREIMS spectrometry, revealed one new amino amidine derivative (1), together with five known simple cyclic dipeptide analogs, diketopiperazines (2-6) and eleven other known compounds, including one hemi-cycline (10), three aromatic derivatives (11-13), one sesquiterpene (14) and three sterols (15-17). The isolated compounds were screened for anticancer and anti-pathogenic bacterial and fungal activities. Based on this work, Xylaria cf. cubensis SWUF08-86 has proven to be a diverse secondary metabolites producer.

A new cerebroside and the cytotoxic constituents isolated from Xylaria allantoidea SWUF76.

A new cerebroside, namely allantoside (1), and 10 known compounds (2-11) were isolated from Xylaria allantoidea SWUF76. The structure of compound 1 was determined by comprehensive spectroscopic analysis including 1D and 2D nuclear magnetic resonance (NMR) as well as high-resolution electron ionisation mass spectrometry (HREIMS) and electrospray ionisation mass spectrometry (ESIMS). Compounds 1, 4, 5, 6, 7, 8 and 11 were evaluated for cytotoxic activities against cancer cell lines (Hela, HT29, HCT116 and MCF-7) and normal Vero cell lines by MTT assay. Compounds 6 and 7 exhibited anticancer activity after 24 h of treatment. Compound 7 showed significant cytotoxicity against Hela (IC50 = 2.24 µg/mL), HT29 (IC50 = 2.51 µg/mL), HCT116 (IC50 = 3.50 µg/mL) and MCF-7 (IC50 = 3.77 µg/mL) and Vero (IC50:3.65 µg/mL) cells. Compound 6 showed slight cytotoxicity against all tested cancer cell lines.

Antimalarial and cytotoxic constituents of Xylaria cf. cubensis PK108.

Xylaria cf. cubensis PK108 was identified by its distinctive morphological characteristics and its internal transcribed spacers sequence analysis. The chromatographic separation and structural elucidation based on spectroscopic analysis of fungal crude extracts led to 10 compounds; tryptoquivaline L (1), fiscalin C (2), epi-fiscalin C (3), cytochalasin D (4), ergosterol (5), ergosterol peroxide (6), chevalone C (7), xylaranol B (8), helvolic acid (9) and cyclo-(L-Pro-L-Leu) (10). The bioassay screening showed that 4 displayed cytotoxicity against KB and NCI-H187 cancer cell lines with IC50 values of 3.25 and 5.95 µg mL(-1). 6 exhibited cytotoxicity against NCI-H187 with an IC50 value of 5.81 µg mL(-1). 7 and 9 showed antimalarial activity with IC50 values of 25.00 and 6.25 µg mL(-1), respectively. This result establishes Xylaria as broad spectrum bioactive compound producers.