Relationship between the Quality of Konjac Flour and the Molecular Matter Nature of Konjac Mannan

We compared the immunomodulating activities in mice of extracts from Phellinus linteus grown on germinated brown rice (PB), Phellinus linteus (PL) alone, and germinated brown rice (BR) alone. The PL, BR and PB-treated mice were administered with the respective extract (2 mg/head/day) by oral gavage for 4 weeks. All extracts markedly decreased the IgE production and allergic responses in serum and splenocytes. PL and PB increased the proportion of CD4⁺ but not CD8⁺ T cells in splenocytes. Cytokine production was significantly augmented in all treated mice; the concentration of IFN-γ was greater in the PL, BR and PB mice than in the control group. The concentration of IL-10 was lower in the BR group than in the other groups. These results may be related to the suppression of IgE production. We conclude that PB modulated the immune responses of IgE production and Th1/Th2 cytokine secretion in murine splenocytes.     

<Emphasis Type="BoldItalic">Phellinus linteus</Emphasis> grown on germinated brown rice suppresses IgE production by the modulation of Th1/Th2 balance in murine mesenteric lymph node lymphocytes

We compared the immunomodulating effects of Phellinus linteus (PL), germinated brown rice (BR) and P.␣linteus grown on germinated brown rice (PB) on IgE production in murine mesenteric lymph node (MLN) lymphocytes. All extracts decreased IgE concentrations by 43–65% compared to control mice in both serum and MLN lymphocytes. In addition, PL and PB increased the proportion of CD4⁺ T cells by␣9% and 12% in MLN lymphocytes. IFN-γ concentration, Th1 cytokine, was significantly increased by 44–67%, whereas IL-4 and IL-10 concentrations, Th2 cytokine, significantly decreased by 30–60% in the three treated groups compared to control group. These results suggest that PB suppresses IgE production through the modulation of Th1/Th2 balance to down-regulate Th2 response in MLN lymphocytes, even though a synergistic effect of PB was not found.     

Fractionation and Anti-tumor Activity of the Mycelia of Liquid-cultured Phellinus linteus

All the fractions of Phellinus linteus mycelia showed anti-tumor activity toward solid tumors planted in mice. The highest anti-tumor activity of 81.2% was observed in the protein–glucan complex obtained by precipitating the 24% NaOH extract at pH 6.0. This protein–glucan complex consisted of 39.3% polysaccharide and 49.4% protein. Its ¹³C- and ¹H-NMR data showed that the main glucan part of the complex was simple α-1,3-glucan chains.     

The Production of β-Fructofuranosidase from Bifidobacterium spp.

The productions of β-fructofuranosidase from Bifidohacterium longum A1, B. adolescentis G1, and four other strains of Bifidobacteria were investigated. All strains used in this study were grown in modified BL broth containing a mixture of fructooligosaccharides [1^F (1-β-D-fructofuranosyl)_n-1sucrose, GF_n (n = 2 – 5)] as the only carbon source. Hydrolyses of 1-kestose, sucrose, and inulin were detected in the extract of the cell. The highest activity on 1-kestose was detected in the extract of B. longum A1 followed by B. adolescentis G1. The other extracts weakly attacked 1-kestose. The relative activities of the extract of B. adolescentis G1 for 1-kestose, nystose, 1^F-fructosylnystose, sucrose, and inulin were 100, 82.5, 50.8, 28.3, and 15.0, respectively. The relative activities for various substrates differed from invertases (yeast β-fructofuranosidases) and exo-inulinase from Penicillium trzehinskii.     

Production of a hypoglycemic, extracellular polysaccharide from the submerged culture of the mushroom, Phellinus linteus

Mycelial growth and extracellular polysaccharide production of Phellinus linteus were optimal at pH 5 and 25 °C. Maximum biomass production (14.2 g l^–1) was after 15 d of cultivation, whereas, extracellular polysaccharide was maximal (3.5 g l^–1) after 21 d. The hypoglycemic effect of the polysaccharide, investigated in streptozotocin-induced diabetic rats, decreased plasma glucose, total cholesterol and triacylglycerol concentrations by 49%, 32%, and 28%, respectively, and aspartate aminotransferase activity by 20%. The results indicate the potential of this polysaccharide to prevent hyperglycemia in diabetic patients.     

Extracts of Phellinus linteus grown on germinated brown rice suppress liver damage induced by carbon tetrachloride in rats

Extracts of Phellinus linteus (EPB), grown on germinated brown rice, protected rats from liver injury induced by carbon tetrachloride (CCl4). Peroxidation products in the liver were decreased to 10% by EPB. Catalase and superoxide dismutase activities were significantly decreased to 55% and 39% by CCl4 administration, but EPB blocked this effect, resulting in enzyme activities at control levels. Expression of cytochromeP450 2E1 (CYP2E1) protein was significantly decreased to 88% in CCl4-treated rats but remained at control levels when EPB was also administered. EPB did not affect the altered fatty acid composition induced by CCl4. The hepatoprotective effect of EPB may be mediated by EPB's prevention of CCl4-induced CYP2E1 degradation.     

Phellinstatin, a new inhibitor of enoyl-ACP reductase produced by the medicinal fungus Phellinus linteus

A new trimeric hispidin derivative, phellinstatin, was isolated from a culture broth of the medicinal fungus Phellinus linteus and its structure was established by various spectral analysis. Phellinstatin strongly inhibited Staphylococcus aureus enoyl-ACP reductase with an IC₅₀ of 6 μM and also showed antibacterial activity against S. aureus and MRSA.     

Identification of metabolites produced from <Emphasis Type="Italic">N</Emphasis>-phenylpiperazine by <Emphasis Type="Italic">Mycobacterium</Emphasis> spp

Mycobacterium sp. 7E1B1W and seven other mycobacterial strains known to degrade hydrocarbons were investigated to determine their ability to metabolize the piperazine ring, a substructure found in many drugs. Cultures were grown at 30°C in tryptic soy broth and dosed with 3.1 mM N-phenylpiperazine hydrochloride; samples were removed at intervals and extracted with ethyl acetate. Two metabolites were purified from each of the extracts by high-performance liquid chromatography; they were identified by mass spectrometry and ¹H nuclear magnetic resonance spectroscopy as N-(2-anilinoethyl)acetamide and N-acetyl-N′-phenylpiperazine. The results show that mycobacteria have the ability to acetylate piperazine rings and cleave carbon-nitrogen bonds.     

Physiological modelling of the response of Kocuria rosea exposed to changing water activity

Physiological effects of NaCl concentration (equivalent water activities, a_w, 1 to 0.87) were investigated with the moderately halophilic and piezotolerant bacterium, Kocuria rosea (formerly Micrococcus roseus), grown in bacteriological peptone/yeast extract broth. This bacterium, which was isolated from open shallow seawater, can grow in 150 g NaCl l^–1 (optimum NaCl concentration: 30 g l^–1, a_w=0.984) and under 207 MPa of hydrostatic pressure. The effects of water activity on _m can be quantitatively predicted, to a high level of accuracy by application of the Aiba/Edwards, and the Levenspiel-type unstructured inhibition-type kinetic models.     

Exploring QSAR for CYP11B2 binding affinity and CYP11B2/CYP11B1 selectivity of diverse functional compounds using GFA and G/PLS techniques

A data set of a series of 132 structurally diverse compounds with cytochrome 11B2 and 11B1 (CYP11B2 and CYP11B1) enzyme inhibitory activities was subjected to molecular shape analysis to explore contributions of shape features as well as electronic, structural, and physicochemical parameters toward enzyme inhibitory activities, in search of appropriate molecular scaffolds with optimum substitutions for highly potent CYP11B2 inhibitors. Genetic function approximation (GFA) and genetic partial least squares (G/PLS) were used as chemometric tools for modeling, and the derived equations were of acceptable statistical quality considering both internal and external validation parameters (Q²: 0.514–0.659, R²_pred: 0.510–0.734). The G/PLS models with spline option for CYP11B2 and CYP11B1 inhibition and selectivity modeling appeared to be the best models based on r_m²_(overall) criterion. The study indicates the importance of the pyridinylnaphthalene and pyridylmethylene-indane scaffolds with less polar and electrophilic substituents for optimum CYP11B2 inhibitory activity and CYP11B2/CYP11B1 selectivity.     

Inhibition of cytochrome P450 isozymes and ornithine decarboxylase activities by polysaccharides from soybeans fermented with Phellinus igniarius or Agrocybe cylindracea

Polysaccharides (5, 10, 25, 50 and 100 microg ml(-1)) from soybeans and soybeans fermented with Phellinus igniarius or Agrocybe cylindracea inhibited cytochrome P450 1A1, cytochrome P450 1A2 and cytochrome P450 2B1 activities in rat liver microsomes. The polysaccharides (5, 10 and 25 microg ml(-1)) also suppressed 12-O-tetradecanoylphorbol-13-acetate-induced ornithine decarboxylase activity. The most potent inhibitors of cytochrome P450 isozymes and ornithine decarboxylase activities were the polysaccharides from soybeans fermented with Agrocybe cylindracea.     

Production of cellulolytic and xylanolytic enzymes by an Arthrographis species

A fungal isolate, Arthrographis sp. strain F4, when grown in shake-flask culture, produced cellulolytic and xylanolytic enzymes optimally at 30°C with an initial pH of 5.0 to 6.0. Coarsely-ground filter paper was the most suitable carbon substrate for production of the enzymes. Inorganic nitrogen sources gave higher activities of the enzymes than organic nitrogen sources: NH₄NO₃ and yeast extract was the most effective combination. Significant stimulation (P<0.05) of enzyme production was achieved with 0.1% (v/v) Tween 80.B.C. Okeke was and S.K.C. Obi is with the Department of Microbiology, University of Nigeria, Nsukka, Nigeria. B.C. Okeke is now with the Department of Bioscience and Biotechnology, Royal College Building, University of Strathclyde, Glasgow G1 1XW, UK     

Pleiotropic consequences of mutations towards antibiotic-hypersensitivity in Serratia marcescens

Various mutants (oxa ^s ) were isolated from Serratia marcescens SM-6 by selecting for hypersensitivity towards oxacillin. All mutants found are highly pleiotropic and able to yield spontaneous revertants which behave like the wild-type. Mutant W 1421 mostly studied shows the following phenotypic properties not found in the wild-type: (1) The growth is hypersensitive to various antibiotics, detergents and dyes which differ remarkably in their chemical structure and antibacterial action-mechanism, (2) the cells can be easily solubilized by 0.05% Sodium-dodecylsulfate, (3) the cells allow the adsorption of the roughmutant specific Salmonella phage 6SR, (4) strong cellular binding of crystal violet, (5) agglutination of the cells in 0.3% auramin solution and (6) reduced formation of red pigment. Strain W 1421 is assumed to be a lipopolysaccharide-defective mutant. The outer membrane of mutant W 1421 analyzed by Sodiumdodecylsulfate-polyacrylamide gel electrophoresis possesses a single protein less than that of the wild-type. Mutant W 1421 is further characterized by its low exolipase activity; exoprotease and exonuclease activities are as in the wild-type. This specific exoenzyme deficiency can be overcome either by backmutation to oxacillin-resistance or by growing mutant W 1421 in a medium supplemented with certain non-metabolizable polysaccharides, e.g. glycogen or pectin B. Both polysaccharides increase the exolipase activity of the wild-type too.List of Abbreviations amp ampicillin - LPS lipopolysaccharide - MIC minimal inhibitory concentration - NB nutrient broth - oxa oxacillin - str streptomycin - TBY tryptone broth with yeast extract - SDS sodium-dodecylsulfate - OD optical density This paper is dedicated to Prof. Dr. R. W. Kaplan, University of Frankfurt/M., on the occasion of his 65th birthday     

Polysaccharides from Phellinus linteus inhibit cell growth and invasion and induce apoptosis in HepG2 human hepatocellular carcinoma cells

It has been demonstrated that the medicinal mushroom Phellinus linteus (PL), which consists mainly of polysaccharides, possesses antitumor and immunomodulatory properties in vivo and in vitro. The mechanism, however, by which PL inhibits growth and invasive behavior of HepG2 cells remains poorly understood. Here we demonstrated that PL inhibited proliferation and colony formation of HepG2 and that the growth inhibition of HepG2 cells was mediated by S-phase cell cycle arrest. PL also markedly inhibited cancer cell adhesion and invasion of the extracellular matrix. Additionally, we demonstrated that PL-induced apoptosis was associated with a reduction in B-cell lymphoma 2 levels and an increase in the release of cytochrome c. These results suggest that PL exerts a direct antitumor effect by initiating apoptosis and cell cycle blockade in HepG2 cells.     

Optimization of Xylanase Production by Thermomyces Lanuginosus in Tomato Seed Meal Using Response Surface Methodology

Summary A 3² central composite experimental design was performed with the aim of optimizing xylanase production by Thermomyces lanuginosus grown on corn cobs in submerged cultures. Xylanase production was first tested on different nitrogen sources (tomato skin, tomato seed meal, corn steep liquor, meat peptone, bacto-tryptone and yeast extract). Tomato seed meal was the selected substrate to test the effect of two variables on xylanase production (corn cobs and tomato seed meal concentrations). A second-order quadratic model and a response surface method showed that the optimum condition for xylanase production was corn cobs 4.6% (w/v) and tomato seed meal 2.1% (w/v). The optimum conditions found were transferred to 7-l bioreactors, where activities as high as 1630 U/ml were obtained.     

Two new benzofuran derivatives from the fungus Stereum sp. YMF1.1684

A basidiomycete fungus, Stereum sp. YMF1.1684, was studied for its ability to produce new compounds on nutrient-enriched cultivation media. One new benzofuran, 2-hydroxy-1-((S)-2-(prop-1-en-2-yl)-2,3-dihydrobenzofuran-5-yl)propan-1-one, named as phenostereum A (1) and one novel dimer benzofuran, named as phenostereum B (2) were isolated from EtOAc extract of the culture broth of the fungus Stereum sp. YMF1.1684. The structures were elucidated using spectroscopic data from 1D, 2D NMR and HRESIMS experiments.     

Tumor-inhibitory and liver-protective effects of <Emphasis Type="Italic">Phellinus igniarius</Emphasis> extracellular polysaccharides

The tumor-inhibitory and liver-protective effects of crude extracellular polysaccharides (EPS) extracted from the liquid mycelial culture of the mushroom Phellinus igniarius were studied in mice. The mice were injected with murine sarcoma S180 and murine hepatoma H22. Crude EPS at 100, 200, 400 mg kg⁻¹ body weight was administered to EPS groups each day in the twelve consecutive days. The result showed that EPS 200 mg kg⁻¹ body weight significantly inhibited S180 and H22 at 65.0 and 46.3%, respectively. Moreover, EPS could not only keep the numbers of WBC, RBC, PLT and the concentration of HGB in a normal range, but also normalize the activities of AST, ALT and ALP. For example, in EPS-treated mice, AST significantly reduced with the percentage of A/G reverse in S180 (P < 0.05) and H22 (P < 0.01) when the mice took EPS 200 mg kg⁻¹ body weight. In conclusion, it was remarkable that P. igniarius EPS exhibited antitumor activity related to dosage and protected liver function by sustaining the blood routine as well as keeping the blood biochemical indexes normal.     

Protein a of quinolinate synthetase is the site of oxygen poisoning of pyridine nucleotide coenzyme synthesis in Escherichia coli

De novo biosynthesis of pyridine nucleotide coenzymes in Escherichia coli is initiated by an enzyme complex (quinolinate synthetase) containing protein B which converts l-aspartate into iminoaspartate protein A, which then generates quinolinate on the pathway to the coenzymes. This complex has been shown to be poisoned by hyperbaric oxygen. ^7,8 We performed assays made dependent on both proteins B and A versus only protein A, using cell-free extracts of hyperbaric-oxygen poisoned and aerobically grown cells. The specific activities were produced by a similar amounts of 68% and 60%, respectively, when measured in assays made dependent on enzymes B and A versus only protein A that was derived from oxygen-poisoned extract. Thus, protein A is the oxygen-sensitive component.     

Emulsification using environmental compatible emulsifiers and de-emulsification using D.C. field and immobilized Nocardia amarae

As environmental compatible emulsifiers, various polysaccharides were investigated and de-emulsification methods were studied using filtration, direct current (D.C.) field, and also Nocardia amarae. 0.01% (v/v) of alginic acid, arabic gum, chitosan, and curdlan showed more than about 2 h of emulsion stability in the half-life of emulsion layer by a homogenizer and showed about twice synergic effects with other polysaccharides in emulsification activity at the ratio of 9:1 (v/v). De-emulsification by 110V D.C. field took about 1 h for the separation of oils from 1 l of oil-in-water emulsion in case of 0.01% arabic gum. Oil-water separator was designed using non-woven fabrics in filtration system and Nocardia amarae grown in n-hexadecane or kerosene was immobilized in the non-woven fabrics.     

Polysaccharides isolated from <Emphasis Type="Italic">Phellinus baumii</Emphasis> stimulate murine splenocyte proliferation and inhibit the lipopolysaccharide-induced nitric oxide production in RAW264.7 murine macrophages

Polysaccharides isolated from Phellinus baumii (PBP) significantly enhanced both lipopolysaccharide (LPS)-induced B lymphocyte proliferation and concanavalin A (Con A)-induced T lymphocyte proliferation. However, PBP (12.5–100 μg ml⁻¹) significantly suppressed the LPS-induced nitric oxide (NO) production in RAW264.7 cells in a concentration-dependent manner. The maximal inhibition of PBP on NO production was 37.5% at 100 μg ml⁻¹. These results provide useful in vitro information to explain the immunostimulating activity and anti-inflammatory activity of PBP.