Trametes versicolor (L.) Lloyd as a source of thermostable serine protease: production and characterization

Proteases are ubiquitously present and are among the largest groups of commercially important enzymes. Here, we investigated a wood-rot basidiomycete Trametes versicolor (L.) Lloyd [Syn. Coriolus versicolor (L.) Quél.; Polyporus versicolor (L.) Fr.] as a source of the enzyme serine protease, its production, and optimized to obtain a higher yield of the enzyme.. The significant variables with optimized values for maximum production of the enzyme were temperature (30?C), incubation time (120 h) and wheat bran (10 g). The yield increased by 30.76% by statistically optimizing the media. The optimized temperature and pH for the maximum protease activity was 50?C and pH 7.0, respectively. The enzyme was purified through ion exchange (using DEAE cellulose 52 resin) and gel filtration chromatography (using Superdex 200 column). The purified enzyme had a retention time of 7 min in RP-HPLC. The enzyme was stable at a broad range of temperature (30-60?C) and pH (5.0-8.0) with a half-life of 58.72 min, Vmax of 37.17 ?M min/mL and Km of 0.657 mg/mL. Its activity was enhanced by Na+, Ca2+, Mg2+ ions and SDS surfactant. These properties make this enzyme a valuable candidate for industrial applications

On the novel inhibitory action of mushroom extract of coriolus versicolor and it’s bioactivity against drug resistant bacteria salmonella typhimurium (MTCC 3241).

The aim of this study was to investigate the effects of the mushroom extract of Coriolus versicolor on drug resistant strain of S. typhimurium experimental results revealed that hot water ethanol extract and Tris- HCl ethanol extract of C. versicolor were not only effective against drug resistant strain of S.typhimurium (MTCC 3214) but also against other pathogenic microorganisms such as K. pneumoniae, S. typhi, E. coli, S. pyogenes and Aspergillus niger. The extracts exerted their effects on the microorganisms by inhibiting their growth.The optimum temperature and pH conditions for mycelial growth of C. versicolor were found to be 28±1ºC and 5.98 pH respectively. Maximum mycelial growth was observed by employing starch and ammonium sulphate as carbon and nitrogen source and on supplementing the cultivation medium with 20% (v/v) culture filtrate of C. versicolor. The present investigation highlights optimization of culture conditions and determination of antimicrobial spectrum C. versicolor .This indicate that the extracts of C. versicolor is a novel inhibitory source against drug resistant bacteria salmonella typhimurium.

Starch degradation and nutrition value improvement in corn grits by solid state fermentation technique with Coriolus versicolor

The study was conducted to evaluate effect of Coriolus versicolor mycelia on degrading starch and improving nutrition value in corn grits through solid state fermentation technique. The results showed that using soybean meal as a nitrogen source, á-amylase secreted from C. versicolor expressed 407.25U/g of activity, leading to 45.15 percent of starch degraded. The activity grew with fermentation time until the 15th day, after that the amylase was deactivated rapidly. An orthogonal experiment designed for the study illustrated that degradation rate of starch in corn grits attained to maximum, 50.51 percent, when 100g of corn grits, added 16g of soybean meal, were fermented by C. versicolor for 12 days, in an initial pH 5.5. After fermenting, compared to the nonfermented control, contents of amino acids, total sugar, crude fat and crude protein were increased by 21.00 percent, 38.45 percent, 55.56 percent, 69.15 percent respectively. The significant improvement of nutrition value in corn grits is probably attributed to the intense metabolism of C. versicolor.

Protective Effect of Coriolus versicolor Cultivated in Citrus Extract Against Nitric Oxide-Induced Apoptosis in Human Neuroblastoma SK-N-MC Cells

Nitric oxide (NO) is a reactive free radical and a messenger molecule in many physiological functions. However, excessive NO is believed to be a mediator of neurotoxicity. The medicinal plant Coriolus versicolor is known to possess anti-tumor and immune-potentiating activities. In this study, we investigated whether Coriolus versicolor possesses a protective effect against NO donor sodium nitroprusside (SNP)-induced apoptosis in the human neuroblastoma cell line SK-N-MC. We utilized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis, and caspase-3 enzyme activity assay in SK-N-MC cells. MTT assay showed that SNP treatment significantly reduces the viability of cells, and the viabilities of cells pre-treated with the aqueous extract of Coriolus versicolor cultivated in citrus extract (CVEcitrus) was increased. However, aqueous extract of Coriolus versicolor cultivated in synthetic medium (CVEsynthetic) showed no protective effect and aqueous citrus extract (CE) had a little protective effect. The cell treated with SNP exhibited several apoptotic features, while those pre-treated for 1 h with CVEcitrus prior to SNP expose showed reduced apoptotic features. The cells pre-treated for 1 h with CVEcitrus prior to SNP expose inhibited p53 and Bax expressions and caspase-3 enzyme activity up-regulated by SNP. We showed that CVEcitrus exerts a protective effect against SNP-induced apoptosis in SK-N-MC cells. Our study suggests that CVEcitrus has therapeutic value in the treatment of a variety of NO-induced brain diseases.

Dye decolorizing potential of a novel fungus Coriolus versicolor ML04 in the medium optimized by response surface methodology

The potential of the white rot fungus, Coriolus versicolor ML04 to decolorize the widely used textile dye Blue BB was tested by employing statistical optimization. Response surface methodology (RSM) involving a central composite design (CCD) was applied to evaluate the interactive effects of four significant factors in different ranges i.e.; glucose (0.5 - 2.5 g/L), yeast extract (0.4 -1.2 g/L), dye concentration (100 - 500 ppm) and inoculum size (5 - 20 percent v/v) to decolorize the Blue BB. The results demonstrated the effectiveness of the statistical experimental design and the ability of C. versicolor ML04 for maximum dye decolorization (>96 percent) at the optimum conditions of the significant factors.

Culture Conditions for Mycelial Growth of Coriolus versicolor

Coriolus versicolor, is one of the most popular medicinal mushrooms due its various biologically active components. This study was conducted to obtain basic information regarding the mycelial culture conditions of C. versicolor. Based on the culture, and MCM media were suitable for the mycelial growth of the mushroom. The optimum carbon and nitrogen sources were dextrin and yeast extract, respectively, and the optimum C/N ratio was 10 to 2 when 2% glucose was used. Other minor components required for optimal growth included thiamine-HCl and biotin as vitamins, succinic acid, lactic acid and citric acid as organic acids, as well as MgSO4.7H2O as mineral salts.

Anti-tumor Effect of Phellinus Linteus and Coriolus Versicolor Capsule on S180 sarcoma and H22 hepatoma in mice / 中国实验方剂学杂志

Objective: To observe the anti-tumor effect of Phellinus Linteus and Coriolus Versicolor Capsules (PLCVC) in S180 sarcoma and H22 hepatoma animal models in mice. Methods: The sarcoma S1180 and hepatoma H22 models were established in mice. After 12 days of treatment, the animals were killed, and the subcutaneous sarcoma were separated and weighted. The levels of vascular endothelial growth factor(VEGF), CD4 and CD8 of S180 tumor tissue were investigated by immunohistochemical method. KM mice were intraperitoneal injected with H22 hepatoma cells, and treated with different experimental drugs. The survival time was observed and recorded, and life-prolongation rate was calculated. Result: PLCVC could inhibit the growth of S180 and H22 tumor, and inhibit the expression of VEGF, improve the expression of CD4 and CD8. The survival time of the mice treated by PLCVC were significantly longer than the untreated group. Conclusion: PLCVC can inhibit the growth of tumour, the mechanism is partially related to inhibiting angiogenesis and improving the immunological function.

Effects of Liquid Culture of Coriolus Versicolor on Lipid Metabolism and Enzyme Activities in Rats Fed High Fat Diet

The effects of liquid culture of Coriolus versicolor on weight gain, food intakes, food efficiency ratios, serum and hepatic lipid concentrations, serum protein levels and serum enzyme activities, were studied in growing male rats. Sprague-Dawley rats were given four different types of diets for a succeeding period of 5 weeks, respectively : a normal diet group (7% corn oil), a high fat diet group (7% corn oil + 15% lard), a 20% or 30% C. versicolor diet groups (high fat diet + 20% or 30% C. versicolor in water) according to the levels of C. versicolor supplementation. The body weight gains of the rats fed the 30% C. versicolor diets were lower than those in the rats fed high fat diet. The epididymal fat pad weight of the rats fed high fat diet and 20% or 30% C. versicolor diets were significantly higher than that of the rats fed normal diet. The concentrations of triglyceride in the serum and the liver of the rats fed the 30% C. versicolor diets were more significantly decreased compared to rats on the high fat diet. The concentrations of total cholesterol in the serum and the liver of rats fed the high fat diet, 20% and 30% C. versicolor diets were similar to those of rats fed the normal diet. The HDL-cholesterol concentration and the HDL-cholesterol/total-cholesterol ratio of the rats fed 20% and 30% C. versicolor diets were significantly lower than those of the rats fed high fat diet. But the atherogenic index of the rats fed 20% or 30% C. versicolor diets were significantly higher than those of the rats fed high fat diet. There were no differences in the activities of glutamic oxaloacetic transaminase, glutamic pyruvic transaminase and alkaline phosphatase in the serum among the experimental groups. These results showed that the 30% C. versicolor diet feeding decreased the triglyceride in serum and liver of the rats.

Ultrastructural Process of Protoplast Fusion Between Lentinula edodes and Coriolus versicolor

Protoplast fusion is a useful technique for establishing fungal hybrids to overcome the natural barriers. The ultrastructure of protoplast and its fusion process were observed using a scanning electron microscopy(SEM) and a transmission electron microscopy(TEM). The protoplasts were variable in size from 0.5~15microm in diameter, and the mean diameter was about 3~5microm. It was impossible to discriminate protoplasts of Lentinula edodes from protoplasts of Coriolus versicolor by size and surface structure. Big aggregates of the dehydrated protoplasts were observed, after polyethylene glycol 4000 treatment. Nucleus, mitochondria, lipid granules and various vesicles having granules were scattered in the cytoplasm. The vesicles were heterogeneous in size and vary from one protoplast to another. The fused membrane layer of the two protoplasts was observed. Time protoplast membrane contact and reorganization of membrane components were essential condition for protoplast fusion. Transmission electron micrograph showed fused protoplasts and flattening of the cells in the area of the membrane contact. We hope that our electron microscopic observations provide some insights into the understanding of the fusion process of protoplast in fungi.

Analgesic Effect of Yunzhi Polysaccharopeptide and Preliminary Analysis of Its Mechanism / 中成药

Objective:To observe the analgesic effect of Yunzhi polysaccharopeptide (PSP) and analyze its mechanism. Methods: The hot plate test in mice and the tail stimulation vocalization test in rats were used.Results: PSP administered po at a dose of 1.0 g/kg for 7 days could produce a significant analgesic effect, which could last for more than two hours. The analgesic effect of PSP disappeared after lesion of mediobasal hypothalamus. Conclusions: PSP could elicit a central analgesic effect.

Purification of Polysaccharide Peptide of Coriolus Versicolor in Affinity Chromatography / 中成药

Objective: To study a new purification method of Polysaccharide Peptide of Coriolus versicolor in affinity chromatography. Methods: Isotherm and kinetics in absorption and the optimal conditions of absorption and elution were studied through static experiments. The static absorption capacity q m and absorption constant K d were calculated according to Chase model. Results: q m =55.57mg/g wet resin, K d =5.312g/L, the dynamic absorption capacity is 43.1mg, polysaccharide/g wet resin and 10.3mg protein/g wet resin.Conclusion: Affinity chromatography can be used to purify PSP preliminarily.