Selenized yeast in production of selenium enriched Pleurotus ostreatus mushroom with good flavour

The aim of this study was to investigate the potential influence of selenized yeast (Sel Plex, Alltech Inc., Lexington, USA) on chemical composition and flavour of solid state grown mushroom Pleurotus ostreatus. Amino acid composition that influences the flavour of selenium-enriched P.ostreatus P80 (137.84 ppm of selenium in d.w.) and non enriched cultivated strains with particular emphasis on selenomethionine was determined by HPLC method, after complete hydrolysis. Volatile flavour compounds of mushroom cultures were analyzed by GC-MS using Headspace sampler. In mushrooms with high selenium content, selenium in the form of L-selenomethionine was present. High selenium concentration in fruit body did not significantly change the amino acid composition of mushrooms. The major amino acids of fruit body sample were glutamic acid, alanine, aspartic acid and tryptophan. In the fruit body of P.ostreatus P80 control, 30 volatile compounds were detected, and in selenium enriched sample number of detected compounds was 25. Compounds that were detected in control sample, but not in eriched fruit body are 2-amino-N-ethylpropanamide, 2-methylpropyl pentan-2-yl sulfite, 2,3-pentanedione, heptan-2-one, pentan-1-ol, 2-methyldihydrofuran-3(2H)-one,1-hydroxyacetone, 3-hydroxy-2-butanon, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine and benzaldehyde. Compounds that were detected in enriched P.ostreatus P80, but not in control are acetone, ethylacetate, 2-propanol, acetonitrile, 2-methylbenzaldehyde and 2-hydroxyethylmethacrylate. Selenized yeast presents good source of selenium for selenium enriched mushroom production with good flavour, which is probably safety for consumption

Optimization of White-Rot Fungi Mycelial Culture Components for Bioremediation of Pharmaceutical-Derived Pollutants

White-rot fungi can degrade a wide spectrum of environmental pollutants, including pharmaceuticals, which are not efficiently removed from wastewater by conventional methods, e.g., the activated sludge method. However, the treatment of wastewater with the use of fungal cultures (mycoremediation) also has significant limitations: among others, the need to use appropriate, often-expensive culture media. We aimed to screen 18 media ingredients, including seven agrifood byproducts for Armillaria mellea, Phanerochaete chrysosporium and Pleurotus ostreatus in submerged cultures to select the low-cost medium optimal for biomass production and laccase activity. We screened nine mathematic models to describe the relation of fungal growth and the amount of the selected byproduct in media. Finally, we tested the ability of the strain with the highest mycelial growth and enzyme-producing ability in the selected medium to degrade eight drug contaminants. Three media variants composed of byproducts provided both efficient growth and laccase production: corn steep liquor + poplar, dried distillers grains with solubles + poplar and corn steep liquor 50%. Among the investigated growth models, the Han–Levenspiel equation described well the specific growth rate in function of the nominal substrate concentration in one-component media. Pleurotus ostreatus, the fungus with the highest ligninolytic enzyme activity, cultured in medium composed of corn steep liquor, removed six of eight drug contaminants with a removal degree of 20–90% in 48 h. The obtained data on the optimal culture media consisting of insoluble components provide initial data for upscaling the process and designing an appropriate type of bioreactor for the process of removing drug contaminants from water

A Search for the Optimum Selenium Source to Obtain Mushroom-Derived Chemopreventive Preparations

The objective of this research was to test whether selenium-yeast (Se-yeast) is a better source of selenium than sodium selenite for accumulation in mycelia and immunoactive cell wall polysaccharides. Culture media were enriched in selenium to a concentration of 20 mu g/mL. Selenium was added to the medium either in the form of sodium selenite or in form of Se-yeast (Sel-Plex; Alltech Inc., Lexington, KY). The total selenium concentrations in the mycelium biomass and in the isolated crude polysaccharides were determined using atomic absorption spectroscopy. We found that selenium accumulated more efficiently in cultures enriched with Se-yeast. A higher concentration of selenium was also found in the crude polysaccharide fractions isolated from the mycelium grown in Se-yeast-enriched media. With the use of the needle trap gas chromatography-mass spectrometry method, we found that there are significant differences in the composition of the volatile aroma and flavor compounds secreted by the mycelia cultivated in different media

Selective Biological Effects of Selenium-Enriched Polysaccharide (Se-Le-30) Isolated from <i>Lentinula edodes</i> Mycelium on Human Immune Cells

A common edible mushroom Lentinula edodes, is an important source of numerous biologically active substances, including polysaccharides, with immunomodulatory and antitumor properties. In the present work, the biological activity of the crude, homogenous (Se)-enriched fraction (named Se-Le-30), which has been isolated from L. edodes mycelium by a modified Chihara method towards human peripheral blood mononuclear cells (PBMCs) and peripheral granulocytes, was investigated. The Se-Le-30 fraction, an analog of lentinan, significantly inhibited the proliferation of human PBMCs stimulated with anti-CD3 antibodies or allostimulated, and down-regulated the production of tumor necrosis factor (TNF)-α by CD3+ T cells. Moreover, it was found that Se-Le-30 significantly reduced the cytotoxic activity of human natural killer (NK) cells. The results suggested the selective immunosuppressive activity of this fraction, which is non-typical for mushroom derived polysaccharides

Identification of the Primary Structure of Selenium-Containing Polysaccharides Selectively Inhibiting T-Cell Proliferation

We previously described the biosynthesis, isolation, and immunosuppressive activity of the selenium-containing polysaccharide fraction isolated from the mycelial culture of Lentinula edodes. Structural studies have shown that the fraction was a protein-containing mixture of high molar mass polysaccharides α- and β-glucans. However, which of the components of the complex fraction is responsible for the immunosuppressive activity non-typical for polysaccharides of fungal origin has not been explained. In the current study, we defined four-polysaccharide components of the Se-containing polysaccharide fraction determined their primary structure and examined the effect on T- and B-cell proliferation. The isolated Se-polysaccharides, α-1,4-glucan (Mw 2.25 × 106 g/mol), unbranched β-1,6-d-glucan, unbranched β-1,3-d-glucan and β-1,3-branched β-1,6-d-glucan (Mw 1.10 × 105 g/mol), are not typical as components of the cell wall of L. edodes. All are biologically active, but the inhibitory effect of the isolated polysaccharides on lymphocyte proliferation was weaker, though more selective than that of the crude fraction

Selenium-Containing Exopolysaccharides Isolated from the Culture Medium of <i>Lentinula edodes</i>: Structure and Biological Activity

In continuation of our research on the influence of selenium incorporation on the biosynthesis, structure, and immunomodulatory and antioxidant activities of polysaccharides of fungal origin, we have isolated from a post-culture medium of Lentinula edodes a selenium (Se)-containing exopolysaccharide fraction composed mainly of a highly branched 1-6-α-mannoprotein of molecular weight 4.5 × 106 Da, with 15% protein component. The structure of this fraction resembled mannoproteins isolated from yeast and other mushroom cultures, but it was characterized by a significantly higher molecular weight. X-ray absorption fine structure spectral analysis in the near edge region (XANES) suggested that selenium in the Se-exopolysaccharide structure was present mainly at the IV oxidation state. The simulation analysis in the EXAFS region suggested the presence of two oxygen atoms in the region surrounding the selenium. On the grounds of our previous studies, we hypothesized that selenium-enriched exopolysaccharides would possess higher biological activity than the non-Se-enriched reference fraction. To perform structure–activity studies, we conducted the same tests of biological activity as for previously obtained mycelial Se-polyglucans. The Se-enriched exopolysaccharide fraction significantly enhanced cell viability when incubated with normal (human umbilical vein endothelial cells (HUVEC)) cells (but this effect was absent for malignant human cervical HeLa cells) and this fraction also protected the cells from oxidative stress conditions. The results of tests on the proliferation of human peripheral blood mononuclear cells suggested a selective immunosuppressive activity, like previously tested Se-polyglucans isolated from L. edodes mycelium. The Se-exopolysaccharide fraction, in concentrations of 10–100 µg/mL, inhibited human T lymphocyte proliferation induced by mitogens, without significant effects on B lymphocytes. As with previously obtained Se-polyglucans, in the currently tested Se-polymannans, the selenium content increased the biological activity. However, the activity of selenium exopolysaccharides in all tests was significantly lower than that of previously tested mycelial isolates, most likely due to a different mode of selenium binding and its higher degree of oxidation

Optimization of Se- and Zn-Enriched Mycelium of <i>Lentinula edodes</i> (Berk.) Pegler as a Dietary Supplement with Immunostimulatory Activity

Mycelial cultures of Lentinula edodes, an edible and medicinal mushroom, have been used in our previous research to obtain selenium-containing immunomodulatory preparations. Our current attempts to obtain a new preparation containing both selenium and zinc, two micronutrients necessary for the functioning of the immune system, extended our interest in the simultaneous accumulation of these elements by mycelia growing in media enriched with selenite and zinc(II) ions. Subsequently, we have studied the effects of new L. edodes mycelium water extracts with different concentrations of selenium and zinc on the activation of T cell fraction in human peripheral blood mononuclear cells (PBMCs). Flow cytometry analysis was used to measure the expression of activation markers on human CD4+ and CD8+ T cells stimulated by anti-CD3 and anti-CD3/CD28 antibodies (Abs). It was demonstrated that statistically significant changes were observed for PD-1 and CD25 antigens on CD8+ T cells. The selenium and zinc content in the examined preparations modified the immunomodulatory activity of mycelial polysaccharides; however, the mechanisms of action of various active ingredients in the mycelial extracts seem to be different