Structural characterization and anti-inflammatory activities of novel polysaccharides obtained from Pleurotus eryngii

Natural polysaccharides named PEP-0.1-1,PEP-0-1 and PEP-0-2 from edible mushroom species Pleurotus eryngii were obtained in the present study.Results showed that molecular weights of these polysaccharides were 3235,2041 and 23933 Da,respectively.Further,structural characterization revealed that PEP-0.1-1 had a→4-α-D-Glcp-1→backbone and contained→4)-α-D-Glcp and→4)-β-D-Glcp reducing end groups.PEP-0-1 backbone contained→4-α-D-Glcp-1→and→6-α-3-O-Me-D-Galp-1→,and the side chains containedα-D-Glcp,β-D-Manp-1→andα-D-Glcp-3→.However,PEP-0-2 backbone consisted of→4-α-DGlcp-1→and→6-α-3-O-Me-D-Galp-(1→6)-α-D-Galp-1→while the side chains containedα-D-Glcp andβ-D-Manp-1→.Biological activity analysis was then carried out and found that all these polysaccharides could significantly suppress the relative mRNA expression of toll-like receptor 4,nitric oxide(NO),tumor necrosis factor-α,interleukin(IL)-1βand IL-6 in lipopolysaccharide(LPS)-induced inflammation of RAW264.7 cells,as well as the over secretion of the above cell cytokines.Moreover,Western blotting analysis revealed that all these purified fractions displayed significant inhibition effects on the expression of c-Jun N-terminal kinases protein induced by LPS in mitogen activated protein kinase pathway,along with the relieving on the inhibition effect of LPS on IκB-αprotein expression.In summary,the information generated by the present study could provide a theoretical basis for the exploration of novel healthy food materials from edible mushroom with antiinflammation activities.

白腐菌Pleurotus eryngii-Co007产木质素降解酶条件的优化

木质素降解酶是参与降解木质素的酶类，广泛存在于白腐真菌中。为提高白腐菌Pleurotus eryngii—Co007的产木质素降解酶的活力，研究了该菌株的产酶曲线、发酵参数及培养基组分对其产木质素降解酶的影响。单因素试验结果表明该菌株发酵第8天时发酵液中木质素降解酶酶活最高；较优的发酵参数是pH5．0—6．0、发酵温度25—27℃，接种量10％～15％，其中pH影响最显著；麦芽糖和有机氮源有利于木质素降解酶的表达。但考虑到酶表达的增加量和生产成本的经济性选择3％的葡萄糖、0．2％酵母粉和0．2％牛肉膏组合氮源；诱导剂中秸杆、Cu^2＋和吐温80诱导产酶效果明显。

Effects of Penicillium spp. and Trichoderma spp.on Pleurotus ostreatus Growth and Screening of Effective Disinfectants

[Objective] This study was conducted to investigate the effects of Penicillium spp. and Trichoderma spp. on the growth of Pleurotus ostreatus （oyster mushroom） and to screen effective disinfectants to control them and other pathogenic microbes. [Method] Six strains or species of Penicilliurn and Trichoderma were inocu- lated into PDA plates growing with P. ostreatus to observe the growth of their mycelia. And the inhibitory effects of hydrogen peroxide, bromogeramine bromide, 84 disinfectant, lysol, potassium permanganate, and 75% medical alcohol on Trichoderma pleuroticola, Trichoderma viride, Trichoderma harzianum, Penicillium citrinum, Penicillium thomii and Penicillium brevicompactum were detected. [Result] Confrontation test revealed that Penicillium spp. and Trichoderma spp. competed for nutrients with P. ostreatus mycelia, antagonized against the growth of P. ostreatus mycelia, invaded and twined around P. ostreatus mycelia, and also produced toxins poisoning P. ostreatus mycelia. The six disinfectants exhibited different inhibitory effects against different species of Penicillium and Trichoderma. Among them the in- hibitory effects of bromogeramine bromide against the six species of Penicillium and Trichoderma were significantly different. However, the inhibitory effects of 3% hydrogen peroxide, 84 disinfectant, 5% lysol, potassium permanganate and 75% medical alcohol were significantly different among Penicillium spp., but did not among Tri- choderma spp. [Conclusion] Bromogeramine bromide can be used to control the bacteria and fungi in mushroom production for it has significant inhibitory effects on the six species of both Penicillium and Trichoderma.

The Interplanting Technology of Pleurotus ostreatus and Dendrobium officinale

In summer, Cha No. 39 of 292 bags and 205 of 244 bags were inter- planted with Dendrobium officinale on cultivation bed, respectively, whose yields and biological efficiencies reached 102.46 kg and 103.98%, and 96.77 kg and 113.32%, accordingly. The seedlings of Dendrobium officinale interplanted with Pleurotus os- treatus grew by 2.78 cm during 10 weeks on cultivation beds, and the seedlings grew by 2.68 cm in the cultivation beds without interplanting arrangement. These demonstrated that the interplanting of Dendrobium officinale and Pleurotus ostreatus has none adverse effects on growth of Dendrobium officinale.

Steroids from the Fungus Pleurotus ostreatus

Twelve steroids, including eight ergostane-type sterols and four mono-glucosides of ergostane-type sterols, were isolated from the ethyl acetate soluble fraction of the fungus Pleurotus ostreatus ( Jacq. : Fr.) Kummer. A new compound 3-O-beta-D-glucopyranosyl-22E, 24R-ergosta-7, 22-diene-5alpha, 6beta, 9alpha-triol, was structurally elucidated by spectroscopic and chemical methods. The eleven known compounds were 22E, 24R-ergosta-7, 22-diene-3beta, 5alpha, 6beta, 9alpha-tetraol; 3-O-beta-D-glucopyranosyl-22E, 24R-ergosta-7, 22-diene-5alpha, 6beta-diol; 22E, 24R-ergosta-7, 22-diene-3beta, 5alpha, 6beta-triol; 22E, 24R-ergosta-5, 7, 22-trien-3beta-ol 3-O-beta-D-glucopyranoside; ergosterol; 22E, 24R-ergosta-7, 22-dien-3beta-ol 3-O-beta-D-glucopyranoside; 22E, 24R-ergosta-7, 22-diene-3beta-ol; 22E, 24R-ergosta-4, 6, 8, 22-tetraen-3-one; 22E, 24R-ergosta-7, 22-diene-3beta, 5alpha, 6alpha-triol; ergosterol peroxide; 22E, 24R-ergosta-7, 22-diene-3beta, 5alpha-diol-6-one.

虎奶菇Pleurotus tuber-regium深层发酵的研究

报道了虎奶菇Pleurotus tuber-regium深层发酵的最适培养基和摇瓶发酵条件。试验结果表明,Pleurotus tuber-regium对单糖、双糖、多糖和糖醇都有一定的利用,但葡萄糖、麦芽糖等对菌丝生长效果较好;酵母粉、黄豆饼粉等蛋白质含量高的氮源对菌丝的生长较为有利;通过L9(34)正交试验确定了Pleurotus tuber-regium发酵的最适培养基为:5%葡萄糖,0.4%酵母粉,0.1%KH2PO4,0.3%MgSO4.7H2O,0.001%VB1及微量NaCl;一定浓度内Na+对菌丝体生长有促进效果。最适的摇瓶发酵条件为:培养基起始pH7.0,培养温度32℃,接种量15%,摇床转速150r/min,250mL三角瓶最适装液量为70mL,发酵周期为6d。

东北林区土著白腐菌Pleurotus ostreatus产漆酶培养基的优化

为提高东北土著白腐真菌的产酶能力,采用摇瓶试验对采自东北林区的白腐真菌Pleurotus ostreatus的产漆酶培养基进行优化,并与目前普遍采用的TK培养基进行对比.结果表明:最佳碳源为玉米粉,质量浓度为8g/L;最佳氮源为氯化铵,质量浓度为0.88g/L.在最佳培养基条件下,白腐真菌Pleurotus ostreatus的漆酶活力和生物量分别达149.45U/L和0.0452g(干重)/L.对比优化后的培养基和目前普遍采用的TK培养基的产漆酶活性,结果表明优化的培养基产漆酶活性是TK培养基的3倍.运用SPSS统计软件对各影响因素进行了差异显著性检验,发现白腐真菌Pleurotus ostreatus产漆酶的主要影响因子为碳源类型.试验证明优化后的培养基能明显促进白腐真菌产漆酶,而选取适合的碳源类型则是配制培养基的关键.

Pleurotus ostreatus对直接耐晒翠蓝染料溶液脱色的研究

研究Pleurotus ostreatus(白腐真菌)对直接耐晒翠蓝GL染料溶液的脱色性能。结果表明,Pleurotus os-treatus能有效用于直接耐晒翠蓝染料溶液的脱色,pH值为5、温度为22℃左右脱色效果最佳;C溶液/C培养基>20%时,增大营养物浓度对脱色率无显著影响;Pleurotus ostreatu对GL的生物吸附符合Langmuir吸附模式,饱和吸附量为20mg/g(湿菌体);并可用Freundlich吸附模型表达,吸附动力学可用准二级速率方程描述;脱色是通过菌体吸附和菌酶降解双重机制实现的。

Quality Evaluation and Standards Development of Liquid Spawn of Pleurotus eryngii

With four Pleurotus eryng# varieties as test materials, the mycelium pelletdensity, dry weight and diameter, fermentation broth pH value, and deoxidized sugar content were determined during their liquid fermentation to investigate the growth of mycelia and changes in fermentation broth. In addition, the sensory, biochemical and microbiological indicators of excellent P. eryngii mycelia were determined by sensory tests, microscope morphology observation and mycelium pellet back-inoculation tests. The results showed that the standards for high-quality liquid spawn of P. eryngii were as follows： bacterial liquid was light yellow and clarifiedwith rich bitter almond flavor but no bacterial contamination; mycelium pellets were visible, white and uniform with burrs on the surface, diameter 〈2.5 mm and concentration of 80%-100%; mycelium pellets were stout, and there were 3--4 clamp connectionsper visual field under the microscope; fermentation time was generally 120-144 h.

Variety Test on Low Temperature-resistant Pleurotus ostreatus Strains in Beijing Area

With 7 familiar Pleurotus ostreatus strains in Beijing region as the test materials,the randomized block design method was adopted to analyze the antagonistic effects,mycelial growth rate,yield of mushroom at low temperature season,total biological efficiency and agronomic characteristics of fruit bodies.The results showed that there were antagonistic effects and differences in all the tested items between all the tested strains,of which the strain PL5 had the shortest spawn age,the strain PL3 showed the highest total biological efficiency,and the fructification of the strain PL7 tasted crisp and tender.

Optimization of the Technique for Liquid Culture of Pleurotus cornucopiae Seeds

[Objective] This study aimed to optimize the conditions for liquid culture of P. comucopiaeseeds. [Method] ＂P. cornucopiae 2＇ selected by our laboratory was used as the experimental strain in this study. Single factor experiment in conjunction with Plackett-Burman （PB） design as well as Response Surface Methodology（RSM） based on Central Composite Design （CCD） were used to screen and optimize the eight factors influencing the technique for liquid culture of P. cornucopiae seeds. [Result] Firstly, the eight factors influencing the biomass of P. cornucopiae seeds in liquid culture were confirmed by means of single factor experiment; secondly, through Plackett-Burman experiment design, three main factors influencing the biomass of P. cornucopiae seeds in liquid culture were screened, namely the amount of added mannitol, the initial pH and culture temperature; lastly, the second order model, that is Y=-70.72＋2.44X3 ＋2.67X7 ＋3.72X8 ＋0.08X3 X7 -0.01X7 X8-0.05X32- 0.34X72-0.08X82, was obtained through Central Composite Design （CCD） along with Response Surface Methodology （RSM）. [Conclusion] The optimal conditions for the production of P. cornucopiae seeds in liquid culture were as follows： the amount of added mannitol of 28.17 g/L, the initial pH controlled at 5.90, 250 ml erlenmeyer flask filled with 100 ml of liquid medium, inoculum size of 5%（v/v）, rotation speed of 150 r/min, and gyratory culture at 23.97℃ for 5 days.

Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii

A versatile peroxidase (VP-Peco60-7 ) was generated and purified from the liquid culture of Pleurotus eryngii. The purification procedure included ammonium sulfate precipitation, ion exchange chromatography, and gel chromatography. The molecular weight and isoelectric point (pI) of VP-Peco60-7 were determined to be approxi-mately 40 kDa and 4.1, respectively. By N-terminal sequence determination and peptide mapping analysis, VP-Peco60-7 was found to be similar to the versatile peroxidase isoenzyme VPL1, which was previously isolated from liquid cultures of the same species. However, the molecular weight and pI of VP-Peco60-7 were different from those of versatile peroxidases of liquid cultures, implying that the VP-Peco60-7 in this study is of a novel type. With 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) as a substrate, the maximal enzyme activity was obtained at 50 °C and pH 3.0. The catalysis of ABTS by VP-Peco60-7 was expressed by the Michaelis-Menten equa-tion. At 50 °C and pH 3.0, the maximum velocity (V max ) was 188.68 U·mg-1 and the michaelis constant (K m ) was 203.09 μmol·L-1 .

小麦秸秆发酵产生的真菌Pleurotus ostreatus IBL-02锰过氧化物酶的溶胶-凝胶固定化及其催化污水脱色(英文)

Solid state bio-processing of wheat straw was carried out through an indigenous fungal strain Pleurotus ostreatus IBL-02 under pre-optimized fermentation conditions. The maximum activity, 692±12 U/mL, of the industrially important manganese peroxidase (MnP) enzyme was recorded after five days of still culture incubation. The crude MnP was 2.1-fold purified with a specific activity of 860 U/mg after purification on a Sephadex-G-100 gel column. On native and SDS-PAGE electrophoresis gels, the purified MnP fraction was a single homogenous band of 45 kDa. An active fraction of MnP was immobilized using hydrophobic sol-gel entrapment comprising tetramethoxysilane (T) and propyltrimethoxysilane (P) at different T:P molar ratios. Characterization revealed that after 24 h incubation at varying pH and temperatures, the MnP fraction immobilized at a T:P ratio of 1:2 in the sol-gel retained 82% and 75% of its original activity at pH4 and 70 ℃, respectively. The optimally active fraction at a 1:2 T:P ratio was tested against MnSO4 as a substrate to determine the kinetic catalytic constants KM and Vmax . To explore the industrial applicability of P. ostreatus IBL-02 MnP, both the free and immobilized MnP were used for the decolorization of four different textile industrial effluents. A maximum of 100% decolorization was achieved for the different textile effluents within the shortest time period. A lower KM , higher Vmax , hyper-activation, and enhanced acidic and thermal resistance up to 70 ℃ were the novel catalytic features of the sol-gel immobilized MnP, suggesting that it may be a potential candidate for biotechnological applications particularly for textile bioremediation purposes.

Protective roles of trehalose in Pleurotus pulmonarius during heat stress response

High temperature is one of the major abiotic stresses that limit edible mushroom growth and development. The understanding of physiological alterations in response to heat stress and the corresponding mechanisms involved is vital for the breeding of heat-resistant edible mushroom strains. Although trehalose functions as a protectant against abiotic stresses in fungi, the putative role of trehalose in thermotolerance remains to be elucidated. In this study, we found heat stress inhibited the growth of two Pleurotus pulmonarius strains, heat-sensitive and less-sensitive, and the inhibition was more significant for the sensitive strain. Heat stress leads to the increase of lipid peroxidation and intracellular trehalose accumulation, with a higher level in the heat-sensitive strain, and this effect is independent of exogenous trehalose application. In addition, a lower concentration of exogenous trehalose application in sensitive strain than in less-sensitive strain was found to alleviate the inhibition of mycelium growth and further increase the intracellular trehalose concentration by heat stress. Thus, the protective effects of trehalose were more remarkable in the sensitive strain. The activities of intracellular trehalose metabolic enzymes, i.e., trehalose-6-phosphate synthase, trehalose phosphorylase and neutral trehalase, were determined, and our data indicated that the changes of these enzymes activities in the sensitive strain were more beneficial to accumulate trehalose than that in the less-sensitive strain.

Application of Pyroligneous Acid in the Prevention and Control of Brown Blotch Disease in Pleurotus ostreatus

In the present study, agar diffusion test and field trials were carried out using a mushroom （Pleurotus ostreatus） culti- var Shouyanping as the experimental material to determine the antibacterial activity of refined pyroligneous acid prepared from almond shell against Pseudomonas tolaasfi and its effects on mycelial growth of mushroom. The results revealed that 1.25%, 2.50% and 5.00% of refined pyroligneous acid obviously inhibited the occurrence of brown blotch disease in mushroom, but also greatly promoted mushroom mycelial growth. Therefore, 1.25% and 2.50% of refined pyroligneous acid can be used in fields to prevent brown blotch disease, and 5.00% refined pyroligneous acid can be used to control the incidence of brown blotch disease when it widely occurs in fields.

Effects of the <i>Camellia oleifera </i>Shell Substrate on the Yield and Nutritional Composition of <i>Pleurotus geesteranus</i>

Pleurotus geesteranus was cultivated on the substrates blended with different ratios of treated and untreated Camellia oleifera shells using cottonseed hull as the control substrate. The mycelial growth rate, yield, nutritional composition, ash and heavy metals of the Pleurotus geesteranus cultivated on these substrates were compared. The results suggest that the Camellia oleifera shell in substrate can accelerate the mycelial growth and increase the yield, nutrients and the contents of protein, ash, crude fiber and amino acid of Pleurotus geesteranus. It was found that the contents of tannin and saponin in Camellia oleifera shell affected the mycelial growth rate. The optimal C/N of the substrate for the growth of Pleurotus geesteranus was determined to be 27 ± 0.7. The C/N ratios higher than 30 reduced the protein, fat and soluble sugar contents of the cultivated Pleurotus geesteranus. The contents of heavy metals including Hg, As, Cd and Pb were found in Pleurotus geesteranus cultivated on the substrates containing Camellia oleifera shell complex.

广泛(尤其在亚洲)种植的凤尾菇(Pleurotus sajor-caju)是一个独立的新种吗

在现代真菌学中,"Pleurotus sajor-caju"具有两种含义:一种是指严格分类学上的含义,即Lentinus sajor-caju的同义命名,另一种是广泛生物技术学含义,即被称为"Pleurotus sajor-caju"的真正隶属于侧耳属(Pleurotus)的一个菌株。文中突出分类地位并给出这一具有疑问的侧耳属成员的正确名称。显微镜分析和BLAST序列比对(内部转录间隔区同源性在97%~100%之间)结果都显示,该问题菌株属于肺形侧耳(P.pulmonarius)。它是肺形侧耳的一个热带生态型,在分类学上是一个新变种(即P.pulmonarius var.stechangii)。该菌的命名是为了铭记卓越的蕈菌学家张树庭教授将这一菌株推广进入食用菌产业。我们对该疑问菌种的先前失真的命名提出更正,并给出了新的学名:Antromycopsis stechangii。

Effect of <i>Carya cathayensis Sarg</i>Shell Substrate on Yield and Nutrient Amount of <i>Pleurotus geesteranus</i>

Using cottonseed hull as the control substrate, different proportions of the untreated and treated Carya cathayensis Sarg shells were added to cultivation of Pleurotus geesteranus. The mycelial growth rate, yield, nutritional composition and contents of heavy metals of the Pleurotus geesteranus cultivated on these substrates were determined. The results suggest that added to treated Carya cathayensis Sarg shell promoted the mycelial growth and increased the yield, biological efficiency and the contents of crude fiber, ash, amino acids and essential amino acids of Pleurotus geesteranus. In addition, the fat content and soluble sugar contents of Pleurotus geesteranus were decreased with the increase of the Carya cathayensis Sarg shell content in substrate, but their contents of heavy metals including mercury (Hg), arsenic (As), cadmium (Cd) and lead (Pb) were higher than that obtained on control substrate.

Bioconversion of low quality lignocellulosic agricultural waste into edible protein by Pleurotus sajor-caju(Fr.) Singer

Pleurotus sajor-caju (Fr.) Singer was cultivated on selected agro wastes viz. cotton stalks, groundnut haulms, soybean straw, pigeon pea stalks; leaves; wheat straw, alone or in combinations. Cotton stalks, pigeon pea stalks; wheat straw alone or in combination were found to be more suitable than groundnut haulms; soybean straw for the cultivation. Organic supplements such as groundnut oilseed cake, gram powder; rice bran not only affected growth parameters but also increased yields. Thus bioconversion of lignocellulosic biomass by P. sajor-caju offers a promising way to convert low quality biomass into an improved human food.

Effects of Different Proportions of Spent Pleurotus ostreatus Substrate on Seed Germination and Seedling Growth of Mung Beans

[Objectives] This study was conducted to investigate the effects of different proportions of spent Pleurotus ostreatus substrate on the germination and seedling growth of mung beans. [Methods] The cellulose-degrading bacteria HB8 and HF1 were mixed with a commercially available microbial composting agent, respectively, for the composting of spent P. ostreatus substrate. Mung beans were cultivated with different proportions of spent mushroom substrate compost and soil. The seed germination rate, plant height, fresh weight and chlorophyll content of mung bean were used as indicators to investigate the effects of the organic fertilizer from spent P. ostreatus substrate on the growth of mung bean seedlings. [Results] The addition of cellulose-degrading bacteria can significantly improve the composting effect of the spent mushroom substrate. After 8 d of cultivation of mung beans with different ratios of the mushroom substrate organic fertilizer, 50% of the organic fertilizer can make the plant height, fresh weight and leaf chlorophyll content of mung bean seedlings reach the highest value and was suitable for mung bean breeding and cultivation. [Conclusions] This study provides scientific basis and technical indicators for the rapid and harmless treatment of spent mushroom substrate and its application in crop cultivation and nursery.