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October 2010

Hydroxyl radical generation by an extracellular low-molecular-weight substance and phenol oxidase activity during wood degradation by the white-rot basidiomycete Trametes versicolor.

One-electron oxidation activity, as measured by ethylene generation from 2-keto-4-thiomethylbutyric acid, phenol oxidase activity, and the generation of hydroxyl radical were examined in cultures of the lignin-degrading white-rot basidiomycete fungus, Trametes (Coriolus) versicolor. The activity levels of specific lignin-degrading enzymes and cellulases, as well as the rate of wood degradation, also were examined. The fungus secreted a low-molecular-weight substance (M(r) 1000-5000) that catalyzed a redox reaction between molecular oxygen and an electron donor, to produce the hydroxyl radical via hydrogen peroxide. During wood decay, T. versicolor also produced significant amounts of laccase and lignin peroxidase, carboxymethyl cellulase, and Avicelase. The roles of the hydroxyl radical, phenol oxidases, and cellulases in wood degradation by white-rot fungi are discussed. That the hydroxyl radical produced by the low-molecular-weight substance secreted by T. versicolor results in new phenolic substructures on the lignin polymer, making it susceptible to attack by laccase or manganese peroxidase is suggested.[…]

Reactions of pentachlorophenol with laccase from Coriolus versicolor.

Laccase, purified from Coriolus versicolor, removed pentachlorophenol (PCP) from solution at pH 5, depending on initial PCP concentration and amount of laccase. With 100 units of laccase, 100% of 25 microg ml(-1) PCP and 60% of 200 microg ml(-1) PCP were removed respectively over 72 h. No free chloride was released in the reaction. In reaction with 100 microg PCP, products were primarily polymers (about 80,000 MW) with only 2-3 pg of o- and p-chloranils formed. Polymers were stable to acid hydrolysis and no release of PCP, or other low-molecular-weight products, was detected over several weeks. Laccase has a potential use in the biotreatment of aqueous effluents containing PCP, with polymerised products being removed from solution due to their high molecular weight.[…]

Effect of environmental conditions on biological decolorization of textile dyestuff by C. versicolor.

Effects of environmental conditions such as pH, media composition, carbon and nitrogen sources, TOC/N ratio, and dyestuff concentrations on decolorization of reactive phytalocyanin type textile dyestuff Everzol Turquoise Blue G by white rot fungi, Coriolus versicolor 20) or low nitrogen concentration was essential for effective decolorization of the dyestuff. Dyestuff concentration should be lower than 500 mg/l for complete decolorization. Only partial decolorization was observed for dyestuff concentrations above 500 mg/l. Adsorption of the dyestuff on surfaces of the fungi was insignificant (<20%).[…]

Generation of nuclear hybrids overcoming the natural barrier of incompatibility: transfer of nuclei from Lentinula edodes into protoplasts of Coriolus versicolor.

Heterokaryotic nuclear hybrids overcoming the natural barriers of incompatibility have been studied in basidiomycetes. To produce these nuclear hybrids between incompatible mushrooms, which have several potent pharmacological effects, nuclear transfer was performed between Lentinula edodes and Coriolus versicolor. Nuclei from serine auxotrophs of Lentinula edodes, LE207 (Ser-) were transferred into the protoplasts of arginine auxotrophs of Coriolus versicolor, CV17 (Arg-), using 30% polyethylene glycol 4000 in 10 mM CaCl2-glycine solution (pH 8.0). Nuclear transfer progenies were selected by nutritional complementation on minimal media supplemented with 0.6 M sucrose. The progenies were classified based on colony morphology to L. edodes-like, C. versicolor-like and non-parental type. Most of the progenies grew slower than either parent. The number of nuclei per cell was similar but the DNA content varied between progenies. The isozyme patterns of nuclear hybrids resembled either of the parent profiles or showed a mixed profile.[…]

Effect of ethanol on enzymatic activity of fungal laccases.

Blue laccase from Coriolus versicolor and blue and yellow laccases from Panus tigrinus were isolated, purified and studied in acetate buffer solutions, with and without addition of various amounts of ethanol, using syringaldazine and 2,6-dimethoxyphenol as substrates. Effect of ethanol on blue laccases could be successfully described using the mixed inhibition model, over the range of 0-2.5 M ethanol concentrations. Yellow laccase from P. tigrinus behaves differently, which may be explained by the presence of some extra molecules in its structure, which possibly stabilize the enzyme and might be exchanged in ethanol solutions.[…]

Susceptibility of natural killer (NK) cells to reactive oxygen species (ROS) and their restoration by the mimics of superoxide dismutase (SOD).

Natural killer (NK) cells are susceptible to reactive oxygen species (ROS), and lose the activity by the effects of ROS. Cancer bearing hosts usually suffer from oxidative stress (OS), and the NK-activity decreases to a significantly lower level than normal controls. Superoxide dismutase (SOD)-mimicking substances, such as protein-bound polysaccharide of Coriolus versicolor (Fr) QUEL (PSK) and iron-chelating chlorine e6-Na (FeCNa), can restore the NK-activity of cancer bearing hosts, when collaborating with catalase. Incorporation of 3H-thymidine by ROS-treated NK-cells is not affected, indicating that these cells are still active in the nucleic acid metabolism. Intraperitoneal administration of anti-Asialo GM1 antibody extinguished the NK-activity. NK-cells affected by ROS lost the adherence to target cancer cells in both in vitro and in vivo. ROS may change the surface charge of NK-cells to anionic, resulting in an inability of adhesion to target cancer cells which usually show the negative surface charge.[…]

Effect of polysaccharide krestin on glutathione peroxidase gene expression in mouse peritoneal macrophages.

Polysaccharide krestin (PSK) is a protein-bound polysaccharide extracted from the sporophore Coriolus versicolor. Previously, we found that PSK could reduce the oxidative injury that oxidised low-density lipoprotein (Ox-LDL) produced in monocytes/macrophages, and therefore have some pro-phylactic or therapeutic effect on atherosclerosis. Glutathione peroxidases, including selenium-dependent glutathione peroxidase (SeGPx) and non-selenium-dependent glutathione peroxidase (non-SeGPx, also called glutathione S-transferase [GST]), play an important role in the defence against oxidative injury. In order to find out if the effects of PSK were associated with antioxidant enzymes, we investigated its effect on glutathione peroxidase activity and messenger RNA (mRNA) expression in mouse peritoneal macrophages. Results showed that PSK enhanced SeGPx and non-SeGPx activity, and increased SeGPx and GST-P (pi class GST) mRNA in mouse peritoneal macrophages. In addition, the induction by PSK of the two glutathione peroxidases could be blocked by cycloheximide (30 micrograms/mL), but 5 micrograms/mL actinomycin D and 50 micrograms/mL acetovanilone (a superoxide inhibitor) had no effect. We conclude that PSK improved glutathione peroxidase activity through transcriptional induction of mRNA expression.[…]

Effect of spent cotton stalks on color removal and chemical oxygen demand lowering in olive oil mill wastewater by white rot fungi.

Wastewater from olive oil mill was decolorized (and its chemical oxygen demand reduced in static cultivation) using the fungi Coriolus versicolor, Funalia trogii, Phanerochaete chrysosporium and Pleurotus sajor-caju. The effect of cotton stalk on decolorizing and COD removing capability was demonstrated. P. chrysosporium (in 20% medium with cotton stalk) reduced the COD by 48% and color by 58%, F. trogii (in 30% medium with cotton stalk)) by 51 and 55%, respectively.[…]

Cell growth and gene modulatory activities of Yunzhi (Windsor Wunxi) from mushroom Trametes versicolor in androgen-dependent and androgen-insensitive human prostate cancer cells.

The incidence of prostate cancer varies greatly throughout the world; it is highest in African-Americans and lowest in the Asian populations of China, India, and Japan. Geographical differences in both prevalence of latent prostate cancer and mortality have been postulated to be influenced by diverse tumor-promoting and protective factors, both environmental and dietary. Prostate cancer is a tumor with an extremely long latency; the pattern of prostate tumorigenesis, in terms of the display and sequence of appearance of particular molecular or biochemical features, or morphological changes, characterizing different stages of the carcinogenic process, is expected to be heterogeneous. Some insights into tumor heterogeneity and progression can be obtained from studies using cell lines, particularly those derived from different anatomical sites. The present study aims to investigate whether hormone-responsive LNCaP and androgen-refractory JCA-1, PC-3, and DU-145 prostate cancer cells are responsive to Yunzhi (YZ), a proprietary dietary supplement prepared from extracts of Trametes versicolor, also known as Coriolus versicolor (a mushroom consumed by Chinese for its purported health benefits), and to elucidate its mechanism of action. Ethanolic extracts (70%) of YZ significantly reduced LNCaP cell growth, down-regulated the levels of secreted PSA, but had less effects on the expression of intracellular PSA and did not affect levels of the androgen receptor. In androgen-unresponsive prostate cancer cells, YZ had a much less pronounced suppressive effect on proliferation of PC-3 and DU-145 cells, compared to LNCaP, and was inactive against JCA-1 cells. Western blot analyses show that the expression of Rb, a key regulatory protein in G1/S transition, and PCNA, integrally involved in mammalian cell DNA replication, were significantly reduced by treatment with YZ in PC-3 and DU-145 cells, respectively. In contradiction, none of these biochemical parameters were affected in JCA-1 cells under identical treatment conditions. Further analysis shows that YZ increased the levels of signal transducer and activator family of transcription factors STAT 1 and STAT 3 in JCA-1 and not LNCaP cells. The greater sensitivity of LNCaP cells to this polysaccharopeptide raises the possibility that YZ may be considered as an adjuvant therapy in the treatment of hormone responsive prostate cancer; additionally, it may have chemopreventive potential to restrict prostate tumorigenic progression from the hormone-dependent to the hormone-refractory state.[…]

Evaluation of solid substrates for enzyme production by Coriolus versicolor, for use in bioremediation of chlorophenols in aqueous effluents.

In the development of a system for the removal of chlorophenols from aqueous effluents, a range of solid substrates for the growth of Coriolus versicolor were investigated. Substrates included wood chips, cereal grain, wheat husk and wheat bran. Suitability for transformation of chlorophenols depended on laccase production by the fungus. The greatest amount of laccase (90% removal of chlorophenols within 100 min.[…]